PR:000003622angiotensin-converting enzyme 2PR:000016456transmembrane protease serine 2PR:P07711cathepsin L1 (human)GO:0019015viral genomePR:000036197viral proteinCL:0000233plateletCHEBI:5054FibrinGO:0005102receptor bindingGO:0061025membrane fusionGO:0075509endocytosis involved in viral entry into host cellGO:0046718viral entry into host cellHP:0004936Venous thrombosisGO:0042730fibrinolysisGO:0072378blood coagulation, fibrin clot formation3occurrence1increased2decreasedSars-CoV-2<p>Virus from the coronaviridae family related to SARS-CoV, 229E, NL63, OC43, HKU1 and MERS.</p>
<p>Transmitted by aerosols</p>
2021-02-23T04:50:402022-09-09T05:09:369606Homo sapiens10090mouse9666Mustela lutreola9685Felis catus9694Panthera tigris9615Canis familiaris9974Manis javanica9541Macaca fascicularis10036Mesocricetus auratus9669Mustela putorius furoWCS_452646Neovison visonWCS_9606humanBinding to ACE2Binding to ACE2Molecular<p><span style="font-size:12px"><span style="font-family:Arial,Helvetica,sans-serif">Angiotensin-converting enzyme 2 (<a href="https://www.genecards.org/cgi-bin/carddisp.pl?gene=ACE2">ACE2</a>) is an enzyme that can be found either attached to the membrane of the cells (mACE2) in many tissues and in a soluble form form (sACE2). </span></span></p>
<p><span style="font-family:Arial,Helvetica,sans-serif"><span style="font-size:12px">A table on ACE2 expression levels according to tissues <em>(Kim et al.)</em></span></span></p>
<table cellspacing="0" class="MsoTableGrid" style="border-collapse:collapse; border:medium none; height:806px; width:1049px">
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<p style="text-align:center"> </p>
</td>
<td style="background-color:#a6a6a6; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">Sample size</span></strong></span></span></p>
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<td style="background-color:#a6a6a6; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">ACE2 mean expression</span></strong></span></span></p>
</td>
<td style="background-color:#a6a6a6; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">Standard deviation of expression</span></strong></span></span></p>
</td>
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<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Intestine</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">51</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">9.50</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.183</span></span></span></p>
</td>
</tr>
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<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Kidney</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">129</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">9.20</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">2.410</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Stomach</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">35</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">8.25</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">3.715</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Bile duct</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">9</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">7.23</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.163</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Liver</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">50</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">6.86</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.351</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Oral cavity</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">32</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">6.23</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.271</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Lung</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">110</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">5.83</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">0.710</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Thyroid</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">59</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">5.65</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">0.646</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Esophagus</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">11</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">5.31</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.552</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Bladder</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">19</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">5.10</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.809</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Breast</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">113</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">4.61</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">0.961</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Uterus</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">25</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">4.37</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.125</span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">Protaste</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:146px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">52</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">4.35</span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; vertical-align:top; width:147px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.905</span></span></span></p>
</td>
</tr>
</tbody>
</table>
<p><span style="font-size:12px"><span style="font-family:Arial,Helvetica,sans-serif"><strong><span style="color:#0070c0">ACE2 receptors in the brain (endothelial, neuronal and glial cells):</span></strong></span></span></p>
<p><span style="font-size:12px"><span style="font-family:Arial,Helvetica,sans-serif"><span style="color:#0070c0">The highest ACE2 expression level in the brain was found in the pons and medulla oblongata in the human brainstem, containing the medullary respiratory centers (Lukiw et al., 2020). High ACE2 receptor expression was also found in the amygdala, cerebral cortex and in the regions involved in cardiovascular function and central regulation of blood pressure including the sub-fornical organ, nucleus of the tractus solitarius, paraventricular nucleus, and rostral ventrolateral medulla (Gowrisankar and Clark 2016; Xia and Lazartigues 2010). The neurons and glial cells, like astrocytes and microglia also express ACE-2. </span></span></span></p>
<p><span style="font-size:12px"><span style="font-family:Arial,Helvetica,sans-serif"><span style="color:#0070c0">In the brain, ACE2 is expressed in endothelium and vascular smooth muscle cells (Hamming et al., 2004), as well as in neurons and glia (Gallagher et al., 2006; Matsushita et al., 2010; Gowrisankar and Clark, 2016; Xu et al., 2017; de Morais et al., 2018) (from Murta et al., 2020). Astrocytes are the main source of angiotensinogen and express ATR1 and MasR; neurons express ATR1, ACE2, and MasR, and microglia respond to ATR1 activation (Shi et al., 2014; de Morais et al., 2018). </span></span></span></p>
<p><span style="font-size:12px"><span style="font-family:Arial,Helvetica,sans-serif"><span style="color:#1abc9c"><strong><em>ACE2 receptors in the intestines</em></strong></span></span></span></p>
<p dir="ltr" style="text-align:justify"><span style="font-size:12px"><span style="font-family:Arial,Helvetica,sans-serif"><span style="color:#1abc9c"><span style="background-color:transparent">The highest levels of ACE2 are found at the luminal surface of the enterocytes, the differentiated epithelial cells in the small intestine, lower levels in the crypt cells and in the colon (Liang et al, 2020; Hashimoto et al., 2012, Fairweather et al. 2012; Kowalczuk et al. 2008). </span></span></span></span></p>
<p dir="ltr" style="text-align:justify"> </p>
<p dir="ltr" style="text-align:justify"> </p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="font-family:Arial,sans-serif"><span style="color:black"><strong><span style="font-size:9.0pt"><span style="font-family:"Times New Roman",serif"><em>In vitro</em> methods supporting interaction between ACE2 and SARS-CoV-2 spike protein</span></span></strong></span></span></span></p>
<p style="text-align:justify"><span style="font-size:12pt"><span style="font-family:Arial,sans-serif"><span style="color:black"><span style="font-size:9.0pt"><span style="font-family:"Times New Roman",serif">Several reports using surface plasmon resonance (SPR) or biolayer interferometry binding (BLI) approaches. to study the interaction between recombinant ACE2 and S proteins have determined a dissociation constant (Kd) for SARS-CoV S and SARS-CoV-2 S as follow,</span></span></span></span></span></p>
<table cellspacing="0" class="Table" style="border-collapse:collapse; width:568px">
<tbody>
<tr>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">Reference</span></strong></span></span></p>
</td>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">ACE2 protein </span></strong></span></span></p>
</td>
<td style="background-color:#f7f7f7; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; height:28px; width:140px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">SARS-CoV S</span></strong></span></span></p>
</td>
<td style="background-color:#f7f7f7; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; height:28px; width:151px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">SARS-CoV2 S</span></strong></span></span></p>
</td>
<td style="background-color:#f7f7f7; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:1px solid black; height:28px; width:128px">
<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">Method</span></strong></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><strong><span style="font-size:9.0pt">Measured Kd</span></strong></span></span></p>
</td>
</tr>
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<p><span style="font-size:11px"><span style="font-family:Arial,Helvetica,sans-serif">doi:<a class="id-link" href="https://doi.org/10.1126/science.abb2507" rel="noopener" target="_blank">10.1126/science.abb2507</a></span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:102px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1–615 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">306–577 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:128px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">SPR</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">325.8 nM</span></span></span></p>
</td>
</tr>
<tr>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1–1208 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">14.7 nM</span></span></span></p>
</td>
</tr>
<tr>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:19px; width:176px">
<p><span style="font-size:11px"><span style="font-family:Arial,Helvetica,sans-serif">doi:<a class="id-link" href="https://doi.org/10.1001/jama.2020.3786" rel="noopener" target="_blank">10.1001/jama.2020.3786</a></span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:102px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">19–615 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">306–527 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:128px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">SPR</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">408.7 nM</span></span></span></p>
</td>
</tr>
<tr>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">319–541 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">133.3 nM</span></span></span></p>
</td>
</tr>
<tr>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:19px; width:176px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><a href="https://elifesciences.org/articles/61390#bib67" style="color:blue; text-decoration:underline"><span style="font-size:9.0pt">Lan et al., 2020</span></a></span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:102px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">19–615 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">306–527 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:128px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">SPR</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">31.6 nM</span></span></span></p>
</td>
</tr>
<tr>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">319–541 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">4.7 nM</span></span></span></p>
</td>
</tr>
<tr>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:19px; width:176px">
<p><span style="font-size:11px"><span style="font-family:Arial,Helvetica,sans-serif">doi:<a class="id-link" href="https://doi.org/10.1016/j.cell.2020.02.058" rel="noopener" target="_blank">10.1016/j.cell.2020.02.058</a></span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:102px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1–614 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">306–575 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:128px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">BLI</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1.2 nM</span></span></span></p>
</td>
</tr>
<tr>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">328–533 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">5 nM</span></span></span></p>
</td>
</tr>
<tr>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:19px; width:176px">
<p><span style="font-size:11px"><span style="font-family:Arial,Helvetica,sans-serif">doi:<a class="id-link" href="https://doi.org/10.1126/science.abb2507" rel="noopener" target="_blank">10.1126/science.abb2507</a></span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:102px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">1–615 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">306–577 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td rowspan="2" style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:128px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">BLI</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">13.7 nM</span></span></span></p>
</td>
</tr>
<tr>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:140px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"> </span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:151px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">319–591 aa</span></span></span></p>
</td>
<td style="background-color:white; border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; width:156px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt">34.6 nM</span></span></span></p>
</td>
</tr>
</tbody>
</table>
<p><span style="font-size:9.0pt"><span style="font-family:"Times New Roman",serif">Pseudo typed vesicular stomatitis virus expressing SARS-CoV-2 S (VSV-SARS-S2) expression system can be used efficiently infects cell lines, with Calu-3 human lung adenocarcinoma epithelial cell line, CaCo-2 human colorectal adenocarcinoma colon epithelial cell line and Vero African grey monkey kidney epithelial cell line being the most permissive (Hoffmann et al., 2020; Ou et al., 2020). It can be measured using a wide variety of assays targeting different biological phases of infection and altered cell membrane permeability and cell organelle signaling pathway. Other assay measured alteration in the levels of permissive cell lines all express ACE2 or hACE2-expressing 293T cell (e.g. pNUO1-hACE2, pFUSE-hIgG1-Fc2), as previously demonstrated by indirect immunofluorescence (IF) or by immunoblotting are associated with ELISA(W Tai et al., nature 2020). To prioritize the identified potential KEs for selection and to select a KE to serve as a case study, further in-silico data that ACE2 binds to SARS-CoV-2 S is necessary for virus entry. The above analysis outlined can be used evidence-based assessment of molecular evidence as a MIE.</span></span></p>
<p> </p>
<p style="text-align:justify"> </p>
UBERON:0000062organCL:0000000cellHighMixedHighAdult, reproductively matureHighDuring development and at adulthoodHighHighHighModerateModerateLow<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">de Morais SDB, et al. Integrative Physiological Aspects of Brain RAS in Hypertension. Curr Hypertens Rep. 2018 Feb 26; 20(2):10.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Gallagher PE, et al. Distinct roles for ANG II and ANG-(1-7) in the regulation of angiotensin-converting enzyme 2 in rat astrocytes. Am J Physiol Cell Physiol. 2006 Feb; 290(2):C420-6.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Gowrisankar YV, Clark MA. Angiotensin II regulation of angiotensin-converting enzymes in spontaneously hypertensive rat primary astrocyte cultures. J Neurochem. 2016 Jul; 138(1):74-85.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Hamming I et al. Tissue distribution of ACE2 protein, the functional receptor for SARS coronavirus. A first step in understanding SARS pathogenesis. J Pathol. 2004 Jun;203(2):631-7.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Jakhmola S, et al. SARS-CoV-2, an Underestimated Pathogen of the Nervous System. SN Compr Clin Med. 2020.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Lukiw WJ et al. SARS-CoV-2 Infectivity and Neurological Targets in the Brain. Cell Mol Neurobiol. 2020 Aug 25;1-8.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Matsushita T, et al. CSF angiotensin II and angiotensin-converting enzyme levels in anti-aquaporin-4 autoimmunity. J Neurol Sci. 2010 Aug 15; 295(1-2):41-5.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Murta et al. Severe Acute Respiratory Syndrome Coronavirus 2 Impact on the Central Nervous System: Are Astrocytes and Microglia Main Players or Merely Bystanders? ASN Neuro. 2020. PMID: 32878468</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Shi A, et al. Isolation, purification and molecular mechanism of a peanut protein-derived ACE-inhibitory peptide. PLoS One. 2014; 9(10):e111188.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:#0070c0">Xia, H. and Lazartigues, E. Angiotensin-Converting Enzyme 2: Central Regulator for Cardiovascular Function. Curr. Hypertens. 2010 Rep. 12 (3), 170– 175</span></span></span></p>
2020-03-02T03:18:472023-04-03T04:03:07SARS-CoV-2 cell entry SARS-CoV-2 cell entry Cellular<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Coronavirus is recognized by the binding of S protein on the viral surface and angiotensin-converting enzyme 2 (ACE2) receptor on the cellular membrane, followed by viral entry via processing of S protein by transmembrane serine protease 2 (TMPRSS2) <span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Hoffmann et al., 2020b).</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif"> ACE2 is expressed on epithelial cells of the lung and intestine, and also can be found in the heart, kidney, adipose, and male and female reproductive tissues </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Lukassen et al., 2020, Lamers et al., 2020, Chen et al., 2020, Jing et al., 2020, Subramanian et al., 2020)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">. </span></span></span></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SARS-CoV-2 is an enveloped virus characterized by displaying spike proteins at the viral surface (Juraszek et al., 2021). Spike is critical for viral entry (Hoffmann et al., 2020b) and is the primary target of vaccines and therapeutic strategies, as this protein is the immunodominant target for antibodies (Yuan et al., 2020, Ju et al., 2020, Robbiani et al., 2020, Premkumar et al., 2020, Liu et al., 2020). Spike is composed of S1 and S2 subdomains. S1 contains the N-terminal (NTD) and receptor-binding (RBD) domains, and the S2 contains the fusion peptide (FP), heptad repeat 1 (HR1) and HR2, the transmembrane (TM) and cytoplasmic domains (CD) (Lan et al., 2020). S1 leads to the recognition of the angiotensin-converting enzyme 2 (ACE2) receptor and S2 is involved in membrane fusion (Hoffmann et al., 2020b, Letko et al., 2020, Shang et al., 2020).</span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">Upon binding to ACE2, the spike protein needs to be activated (or primed) through proteolytic cleavage (by a host protease) to allow membrane fusion. Fusion is a key step in viral entry as it is the way to release SARS-CoV-2 genetic material inside the cell. Cleavage happens between its spike’s S1 and S2 domains, liberating S2 that inserts its N-terminal domain into a host cell membrane and mediates membrane fusion </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Millet and Whittaker, 2018)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">.</span></span></span> Many proteases were identified to activate coronaviruses including furin, cathepsin L, trypsin-like serine proteases TMPRSS2, TMPRSS4, TMPRSS11, and human airway trypsin-like protease (HATs). These may operate at four different stages of the<a href="https://www.wikipathways.org/index.php/Pathway:WP4846"> virus infection cycle</a>: (a) pro-protein convertases (e.g., furin) during virus packaging in virus-producing cells, (b) extracellular proteases (e.g., elastase) after virus release into extracellular space, (c) cell surface proteases [e.g., type II transmembrane serine protease (TMPRSS2)] after virus attachment to virus-targeting cells, and (d ) lysosomal proteases (e.g., cathepsin L) after virus endocytosis in virus-targeting cells (Li, 2016).<span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif"> SARS-CoV-2 lipidic envelope may fuse with two distinct membrane types, depending on the host protease(s) responsible for cleaving the spike protein: (i) cell surface following activation by serine proteases such as TMPRSS2 and furin </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Hoffmann et al., 2020b)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">; or (ii) endocytic pathway within the endosomal–lysosomal compartments including processing by lysosomal cathepsin L </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Yang and Shen, 2020)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">. These flexibility for host cell factors mediating viral entry, highlights that the availability of factors existing in a cell type dictates the mechanism of viral entry </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Kawase et al., 2012)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">. When TMPRSS2 (or other serine proteases such as TMPRSS4 </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Zang et al., 2020)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif"> or human airway trypsin-like protease [HAT]</span></span></span> <span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Bestle et al., 2020a)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">) is expressed, fusion of the virus with the cell surface membrane is preferred </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Shirato et al., 2018)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">, while in their absence, the virus can penetrate the cell by endocytosis </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Kawase et al., 2012)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">. A third factor has also been shown to facilitate SARS-CoV-2 entry in cells that have ACE2 and even promote, although to very low levels, SARS-CoV-2 entry in cells that lack ACE2 and TMPRSS2 which is the neuropilin-1 (NRP-1) </span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">(Cantuti-Castelvetri et al., 2020)</span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-family:"Calibri",sans-serif">. This key event deals with SARS-CoV-2 entry in host cells and is divided in three categories: TMPRSS2, capthesin L and NRP-1.</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>TMPRSS2 Spike cleavage:</strong></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">TMPRSS2 (transmembrane serine protease 2, (<a href="https://www.ncbi.nlm.nih.gov/gene/7113" style="color:blue; text-decoration:underline">https://www.ncbi.nlm.nih.gov/gene/7113</a>) is a cell-surface protease (Hartenian et al., 2020) that facilitates entry of viruses into host cells by proteolytically cleaving and activating viral envelope glycoproteins. Viruses found to use this protein for cell entry include Influenza virus and the human coronaviruses HCoV-229E, MERS-CoV, SARS-CoV and SARS-CoV-2 (COVID-19 virus).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">TMPRSS2 is a membrane bound serine protease also known as epitheliasin. TMPRSS2 belongs to the S1A class of serine proteases alongside proteins such as factor Xa and trypsin. Whilst there is evidence that TMPRSS2 autoclaves to generate a secreted protease, its physiological function has not been clearly identified. However, it is known to play a crucial role in facilitating entry of coronavirus particles into cells by cleaving the spike protein (Huggins, 2020).</span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">After ACE2 receptor binding, SARS-CoV-2 S proteins can be subsequently cleaved and activated by host cell-surface protease at the S1/S2 and S2’ sites, generating the subunits S1 and S2 that remain non-covalently linked. Cleavage leads to activation of the S2 domain that drives fusion of the viral and host membranes (Hartenian et al., 2020, Walls et al., 2016). For other coronaviruses, processing of spike was proposed to be sequential with S1/S2 cleavage preceding that of S2. Cleavage at S1/S2 may be crucial for inducing conformational changes required for receptor binding or exposure of the S2 site to host proteases. </span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The S1/S2 site of SARS-CoV-2 S protein contains an insertion of four amino acids providing a minimal furin cleavage site (RRAR685↓) (that is absent in SARS-CoV). Interestingly, the furin cleavage site has been implicated in increased viral pathogensis (Bestle et al., 2020b, Huggins, 2020). <span style="color:black">Processing of the spike protein by furin at the S1/S2 cleavage site is thought to occur following viral replication in the endoplasmic reticulum Golgi intermediate compartment (ERGIC) </span><span style="color:black">(Hasan et al., 2020)</span><span style="color:black">. T</span>he spike S2’ cleavage site of SARS-CoV-2 possesses a paired dibasic motif with a single KR segment (KR815↓) (as SARS-CoV) that is recognized by trypsin-like serine proteases such as TMPRSS2. <strong><span style="color:black">The current data support a model for SARS-CoV-2 entry in which furin-mediated cleavage at the S1/S2 site pre-primes spike during biogenesis, facilitating the activation for membrane fusion by a second cleavage event at S2’ by TMPRSS2 following ACE2 binding</span></strong> <span style="color:black">(Bestle et al., 2020b, Johnson et al., 2020)</span><span style="color:black">.</span></span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Virus</span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">S1/S2 site</span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">S2’ site</span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SARS-CoV-2</span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">TNSP<strong>RRAR</strong>|SVA</span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">PSKPS<strong>KR</strong>|SFIEDL</span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SARS-CoV </span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">S----LLR|STS</span></span></p>
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<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">PLKPT<strong>KR</strong>|SFIEDL</span></span></p>
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<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Camostat mesylate, an inhibitor of TMPRSS2, blocks SARS-CoV-2 infection of lung cells like <span style="color:black">Calu-3 cells but not Huh7.5 and Vero E6 cells</span>. Cell entry was assessed using a viral isolate and viral pseudotypes (artificial viruses) expressing the COVID-19 spike (S) protein. The ability of the viral pseudotypes (expressing S protein from SARS-CoV and SARS-CoV-2) to enter human and animal cell lines was demonstrated, showing that SARS-CoV-2 can enter similar cell lines as SARS-CoV. Amino acid analysis and cell culture experiments showed that, like SARS-CoV, SARS-CoV-2 spike protein binds to human and bat angiotensin-converting enzyme 2 (ACE2) and uses a cellular protease TMPRSS2 for priming. Priming activates the spike protein to facilitate viral fusion and entry into cells. Cell culture experiments were performed using immortalized cell lines and primary human lung cells (Hoffmann et al., 2020b, Rahman et al., 2020).</span></span></p>
<p> </p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>Spike binding to neuropilin-1:</strong></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Neuropilin-1 (NRP1) is a transmembrane glycoprotein that serves as a cell surface receptor for semaphorins and various ligands involved in angiogenesis in vertebrates. NRP1 is expressed in neurons, blood vessels (endothelial cells), immune cells and many other cell types in the mammalian body (maternal fetal interface) and binds a range of structurally and functionally diverse extracellular ligands to modulate organ development and function (Raimondi et al., 2016). NRP1 is well described as a co-receptor for members of the class 3 semaphorins (SEMA3) or vascular endothelial growth factors (VEGFs) (Gelfand et al., 2014). Structurally, NRP1 comprises seven sub-domains, of which the first five are extracellular; two CUB domains (a1 and a2), two coagulation factor V/VIII domains (FV/VIII; b1 and b2) and a meprin, A5 μ-phosphatase domain (MAM; c). NRP1 contains only a short cytosolic tail with a PDZ-binding domain lacking internal signaling activity. The different ligand families bind to different sites of NRP1; SEMA3A binding requires the first three sub-domains of NRP1 (a1, a2, and b1), whereas binding of VEGF-A requires the b1 and b2 domains (Muhl et al., 2017). Additional studies conducted by means of in silico computational technology to identify and validate inhibitors of the interaction between NRP1 and SARS-CoV-2 Spike protein are reported in (Perez-Miller et al., 2020). Represents a schematic picture of VEGF-A triggered phosphorylation of VEGF-R2. Screening of NRP-1/VEGF-A165 inhibitors by in-cell Western (Perez-Miller et al., 2020).v NRP1 acts as a co-receptor for SARS-CoV-2. </span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">NRP1 is a receptor for <span style="color:black">furin-cleaved SARS-CoV-2 spike peptide </span><span style="color:black">(Cantuti-Castelvetri et al., 2020, Daly et al., 2020, Johnson et al., 2020)</span><span style="color:black">. Blockade of NRP1 reduces infectivity and entry, and alteration of the furin site leads to loss of NRP1 dependence, reduced replication in Calu3, augmented replication in Vero E6, and attenuated disease in a hamster pathogenesis disease model </span><span style="color:black">(Johnson et al., 2020)</span><span style="color:black">.</span> In fact, a small sequence of amino acids was found that appeared to mimic a protein sequence found in human proteins that interact with NRP1. The spike protein of SARS-CoV-2 binding with NRP1 aids viral infection of human cells. This was confirmed by applying a range of structural and biochemical approaches to establish that the spike protein of SARS-CoV-2 does indeed bind to NRP1. The host protease furin cleaves the full-length precursor S glycoprotein into two associated polypeptides: S1 and S2. Cleavage of S generates a polybasic RRAR C-terminal sequence on S1, which conforms to a C-end rule (CendR) motif that binds to cell surface neuropilin-1 (NRP1) and neuropilin-2 (NRP2) receptors. It was reported that the S1 CendR motif directly bound NRP1 by X-ray crystallography and biochemical approaches. Blocking this interaction using RNAi or selective inhibitors reduced SARS-CoV-2 entry and infectivity in cell culture (Daly et al., 2020).</span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">NRP1, known to bind furin-cleaved substrates, significantly potentiates SARS-CoV-2 infectivity, which was revealed by a monoclonal blocking antibody against NRP1. It was found that a SARS-CoV-2 mutant with an altered furin cleavage site did not depend on NRP1 for infectivity. Pathological analysis of olfactory epithelium obtained from human COVID-19 autopsies revealed that SARS-CoV-2 infected NRP1-positive cells faced the nasal cavity (Cantuti-Castelvetri et al., 2020). Furthermore, it has been found that NRP1 is a new potential SARS<span style="font-family:"Cambria Math",serif">‑</span>CoV<span style="font-family:"Cambria Math",serif">‑</span>2 infection mediator implicated in the neurologic features and central nervous system involvement of COVID<span style="font-family:"Cambria Math",serif">‑</span>19. Preclinical studies have suggested that NRP1, a transmembrane receptor that lacks a cytosolic protein kinase domain and exhibits high expression in the respiratory and olfactory epithelium, may also be implicated in COVID<span style="font-family:"Cambria Math",serif">‑</span>19 by enhancing the entry of SARS<span style="font-family:"Cambria Math",serif">‑</span>CoV<span style="font-family:"Cambria Math",serif">‑</span>2 into the brain through the olfactory epithelium. NRP1 is also expressed in the CNS, including olfactory<span style="font-family:"Cambria Math",serif">‑</span>related regions such as the olfactory tubercles and paraolfactory gyri. Supporting the potential role of NRP1 as an additional SARS<span style="font-family:"Cambria Math",serif">‑</span>CoV<span style="font-family:"Cambria Math",serif">‑</span>2 infection mediator implicated in the neurologic manifestations of COVID<span style="font-family:"Cambria Math",serif">‑</span>19. Accordingly, the neurotropism of SARS<span style="font-family:"Cambria Math",serif">‑</span>CoV<span style="font-family:"Cambria Math",serif">‑</span>2 via NRP1<span style="font-family:"Cambria Math",serif">‑</span>expressing cells in the CNS merits further investigation (Davies et al., 2020).</span></span></p>
<p style="text-align:justify"> </p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Up-regulation of NRP1 protein in diabetic kidney cells hints at its importance in a population at risk of severe COVID-19. Involvement of NRP-1 in immune function is compelling, given the role of an exaggerated immune response in disease severity and deaths due to COVID-19. NRP-1 has been suggested to be an immune checkpoint of T cell memory. It is unknown whether involvement and up-regulation of NRP-1 in COVID-19 may translate into disease outcome and long-term consequences, including possible immune dysfunction (Mayi et al., 2021).</span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The main feature of NRP1 co-receptor is to form complexes with multiple other receptors. Hence, there is a competition between receptors to complex with NRP-1, which may determine their abilities both quantitatively and qualitatively to transduce signals. It is tempting to hypothesize that the occupancy of NRP-1 with one receptor may thus decrease its availability for virus entry. Recent proteomics work has shown that NRP-1 can form a complex with the α7 nicotinic receptor in mice. Both receptors are expressed in the human nasal and pulmonary epithelium (Mayi et al., 2021).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">NRP1, is highly expressed in the respiratory and olfactory epithelium; it is also expressed in the CNS, including olfactory related regions such as the olfactory tubercles and paraolfactory gyri (Davies et al., 2020).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">More information on tissue distribution and protein expression of NRP1 can be found in https://www.proteinatlas.org/ENSG000000992 50-NRP1</span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>Spike entry via <span style="color:black">lysosomal cathepsins and endocytosis</span>:</strong></span></span></p>
<p style="text-align:justify"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:black">Evidence shows the role of TMPRSS2 and other serine proteases in activating the coronavirus spike protein for plasma membrane fusion. However, studies using various cell culture systems showed that SARS-CoV2 could enter cells via an alternative endosomal–lysosomal pathway.</span> Evidence came from studies<span style="color:black"> demonstrating that lysosomotropic agents reduced SARS-CoV replication in cells lacking TMPRSS2 and other studies, using highly potent and specific small-molecule cathepsin inhibitors, to understand the role of cathepsins in processing and activating the spike for membrane fusion, mainly of cathepsin L (one of the 11 cathepsins) </span><span style="color:black">(Rossi et al., 2004, Simmons et al., 2005)</span><span style="color:black">. SARS-CoV-2 and other coronaviruses can establish infection through endosomal entry in commonly used in vitro cell culture systems. Of relevance, inhibitors of the endosomal pathway, as the cathepsin inhibitor Z-FA-FMK and PIKfyve inhibitor apilimod, blocked viral entry in Huh7.5 and Vero E6 cells but not Calu-3 cells.</span></span></span></p>
<p style="text-align:justify"><strong>Viral entry leads to delivery of virion proteins and translation of viral proteins immediately: </strong></p>
<p style="text-align:justify"><span style="font-size:14px">Coronavirus is a class of viruses that have single-stranded positive-sense RNA genomes in their envelopes [Kim D,<em> et al., 2020</em>]. The virus contains a <span style="color:#131413">29.7 kB positive-sense RNA genome flanked by 5' and 3' untranslated regions of 265 and 342 nucleotides, respectively</span><span style="color:black"> </span><span style="color:#131413">that contain cis-acting secondary RNA structures essential for RNA synthesis [</span>Huston N. C.<em> et al., 2021</em>]<span style="color:black">. T</span>he genome just prior to the 5′ end contains the transcriptional regulatory sequence leader (TRS-L) [Budzilowicx C.J., <em>et al., 1985</em>]. The SARS-CoV genome is polycistronic and contains 14 open reading frames (ORFs) that are expressed by poorly understood mechanisms [Snijder E. J., <em>et al.</em>, 2003]<span style="color:black">.</span> Preceding each ORF there are other TRSs called the body TRS (<span style="color:black">TRS B). </span>The <span style="color:black">5′ two-thirds of the </span>genome contains <span style="color:black">two large, overlapping, nonstructural ORFs and the 3′ third contains the remainder ORFs [Di H., <em>et al.</em>, 2018].</span> Genome expression starts with the translation of <span style="color:#131413">two large ORFs of the 5’ two-thirds: ORF1a of</span><span style="color:black"> 4382 amino acids and ORF1ab of 7073 amino acid that occurs via a</span><span style="color:#131413"> programmed (- 1) ribosomal frameshifting </span>[Snijder E. J., <em>et al.</em>, 2003]<span style="color:black">, yielding</span><span style="color:#131413"> pp1a and pp1ab</span><span style="color:black">. These two polyproteins are cleaved into 16 subunits by two viral proteinases encoded by ORF1a,</span> <span style="color:black">nsp3, and nsp5 that contain a papain-like protease domain and a 3C-like protease domain</span> [Sacco M. D. <em>et al., 2020</em>]<span style="color:#131413">. </span><span style="color:black">The processing products are a group of replicative enzymes, named nsp1-nsp16, that assemble into a viral replication a</span>nd transcription <span style="color:black">complex (RTC) associated with membranes of endoplasmic reticulum (ER) with the help of various membrane-associated viral proteins [</span>Klein<em> </em>S., <em>et al., 2021</em>, Snijder E. J.<em>, et al., 2020, </em>V'Kovski P. , <em>et al., 2021</em>]<span style="color:black">. This association leads to replication factories or organelles, that are originate new membranous structures that are observed by electron mciroscopy . They are a feature of all coronaviridae and the site of viral replication and transcription hidden from innate immune molecules.</span></span></p>
<p>SARS-CoV2 entry can be determined by many different ways:</p>
<p>1) quantitative RT-PCR specific to the subgenomic mRNA of the N transcript, in cells manipulated with host factors that express of not TMPRSS2, cathepsinL, neuropilin-1, hACE2 [Glowacka I, et al. (2011)], or exogenous addition of HAT or furin.</p>
<p>2) using spike-pseudotyped viral particles expressing GFP/luciferase/bgalactosidase and comparing with vesicular stomatitis virus G seudotyped particles expressing the same reporter analysed in manipulated cultured with cell lines, followed by determining fluorescence, biolumincescence, luciferase activity in cell lysates [Hoffmann M, et al. (2020)].</p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>TMPRSS2:</strong></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">TMPRSS2 gene expression can be measured by RNAseq and microarray (Baughn et al., 2020).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Expression levels of TMPRSS2 can be measured by RNA in situ hybridization (RNA-ISH) (Qiao et al., 2020)</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>NRP-1:</strong></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Several methods have been identified in the literature for measuring and detecting NRP1 receptor binding. Briefly described:</span></span></p>
<ol>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><span style="color:black">X-ray crystallography and biochemical approaches help to show that the S1 CendR motif directly bound NRP1 (1). Binding of the S1 fragment to NRP1 was assessed and ability of SARS-CoV-2 to use NRP1 to infect cells was measured in angiotensin-converting enzyme-2 (ACE-2)-expressing cell lines by knocking out NRP1 expression, blocking NRP1 with 3 different anti-NRP1 monoclonal antibodies, or using NRP1 small molecule antagonists </span><span style="color:black">(Centers for Disease Control and Prevention, 2020, Daly et al., 2020)</span><span style="color:black">.</span></span></span></li>
</ol>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Key findings (Centers for Disease Control and Prevention, 2020, Daly et al., 2020): </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">• The S1 fragment of the cleaved SARS-CoV-2 spike protein binds to the cell surface receptor neuropilin-1 (NRP1). </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">• SARS-CoV-2 utilizes NRP1 for cell entry as evidenced by decreased infectivity of cells in the presence of: NRP1 deletion (p <0.01). Three different anti-NRP1 monoclonal antibodies (p <0.001). Selective NRP1 antagonist, EG00229 (p <0.01).</span></span></p>
<ol start="2">
<li><span style="font-size:11pt"><span style="color:black"><span style="font-family:"Calibri",sans-serif">Cell lines were modified to express ACE2 and TMPRSS2, the two known SARS-CoV-2 host factors, and NRP1 to assess the contribution of NRP1 to infection. Autopsy specimens from multiple airway sites were stained with antibodies against SARS-CoV-2 proteins, ACE2, and NRP1, to visualize co-localization of proteins (6, 15).</span></span></span></li>
</ol>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Key findings (Cantuti-Castelvetri et al., 2020, Centers for Disease Control and Prevention, 2020): </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">• Infectivity of cells expressing angiotensin converting enzyme-2 (ACE2, receptor for SARS-CoV-2), transmembrane protease serine-2 (TSS2, primes the Spike [S] protein), and neuropilin-1 (NRP1) with pseudovirus expressing the SARS-CoV-2 S1 protein was approximately 3-fold higher than in cells expressing either ACE2 or TSS2 alone (p<0.05).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">• Analysis of autopsy tissue from COVID-19 patients showed co-localization of the SARS-CoV-2 spike (S) protein and NRP1 in olfactory and respiratory epithelium.</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><span style="color:black">Virtual screen of nearly 0.5 million compounds against the NRP-1 CendR site, resulting in nearly 1,000 hits. A pharmacophore model was derived from the identified ligands, considering both steric and electronic requirements. Preparation of receptor protein and grid for virtual screening, docking of known NRP-1 targeting compounds, ELISA based NRP1-VEGF-A165 protein binding assay; more details on methodology in the referenced paper </span></span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif"><span style="color:black">(Perez-Miller et al., 2020)</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">TMPRSS2 vertebrates (Lam et al., 2020)</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">NRP1 in human & rodents (but also present in monkey and other vertebrates </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(Lu and Meng, 2015)</span></span></p>
<p><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">The ability for SARS-CoV-2 to use multiple host pathways for viral entry, means that it is critical to map which viral entry pathway is prevalent in specific cell types. This is key for understanding coronavirus biology, but also use informed decisions to select cells for cell-based genetic and small-molecule screens and to interpret data. In fact, a combination of protease inhibitors that block both TRMPSS2 and cathepsin L is the most efficient combination to block coronavirus infection </span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(Yamamoto et al., 2020, Shang et al., 2020, Shirato et al., 2018)</span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. In accordance, SARS-CoV-2 entry processes are highly dependent on endocytosis and endocytic maturation in cells that do not express TMPRSS2, such as VeroE6 or 293T cells </span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(Murgolo et al., 2021, Kang et al., 2020, Mirabelli et al., 2020, Riva et al., 2020)</span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. However, even in these cells, heterologous expression of TMPRSS2 abrogates the pharmacological blockade of cathepsin inhibitors </span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(Kawase et al., 2012, Hoffmann et al., 2020a)</span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. Treatment of SARS-CoV-2 with trypsin enables viral cell surface entry, even when TMPRSS2 is absent. Moreover, TMPRSS2 is more efficient to promote viral entry than cathepsins </span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(Lamers et al., 2020)</span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">, as when both factors are present,d cathepsin inhibitors are less effective than TMPRSS2 inhibitors </span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(Hoffmann et al., 2020b)</span></span></span></span><span style="font-family:"MinionPro-Regular",serif"><span style="color:black"><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">. Therefore it is critical to map which cells contain the different types of proteases.</span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">In summary, TMPRSS2 appears to be expressed in a wide range of healthy adult organs, but in restricted cell types, including:</span></span></p>
<ul>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">AT2 and clara cells of lungs</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">sinusoidal endothelium, and hepatocyte of the liver, </span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">endocrine cells of the prostate, </span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">goblet cells , and enterocytes of the small intestine, </span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">intercalated cells, and the proximal tubular of the kidney.</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Ciliated, secretory and suprabasal of nasal</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">spermatogonial stem cells of testes</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">cyto tropoblast and peri vascular cells of placenta</span></span></li>
<li><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The nasal epithelium expresses various combinations of factors that, in principle, could facilitate SARS-CoV-2 infection, but it also expresses robust basal levels of RFs, which may act as a strong protective barrier in this tissue.</span></span></li>
</ul>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">There is a shift in TMPRSS2 regulation during nasal epithelium differentiation in young individuals that is not occurring in old individuals (Lin et al., 1999, Lucas et al., 2008, Singh et al., 2020). </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Only a small minority of human respiratory and intestinal cells have genes that express both ACE2 and TMPRSS2. Amongst the ones that do, three main cell types were identified: A) lung cells called type II pneumocytes (which help maintain air sacs, known as alveoli); B) intestinal cells called enterocytes, which help the body absorb nutrients; and C) goblet cells in the nasal passage, which secrete mucus (Ziegler et al., 2020). </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The clinical manifestations of COVID‐19 include not only complications from acute myocardial injury, elevated liver enzymes, and acute kidney injury in patients presenting to hospitals, but also gastrointestinal symptoms in community patients experiencing milder forms of the disease (Madjid et al., 2020, Pan et al., 2020). </span></span></p>
<p> </p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>NRP-1:</strong></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">All life stages</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The expression of isoforms 1 (NRP1) and 2 (NRP2) does not seem to overlap. Isoform 1 is expressed by the blood vessels of different tissues. In the developing embryo it is found predominantly in the nervous system. In adult tissues, it is highly expressed in heart and placenta; moderately in lung, liver, skeletal muscle, kidney and pancreas; and low in adult brain. Isoform 2 is found in liver hepatocytes, kidney distal and proximal tubules. Expressed in colon and 234 other tissues with Low tissue specificity (UniProtKB). </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The expression of NRP1 protein in gastric cancer was not related to gender or age (Cao et al., 2020).</span></span></p>
<p> </p>
<p><strong><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">Sex Applicability:</span></span></strong></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>TMPRSS2:</strong></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Androgen receptors (ARs) play a key role in the transcription of TMPRSS2 (Fig. 1). This may explain the predominance of males to COVID-19 infection, fatality, and severity because males tend to have a higher expression and activation of ARs than females, which is due to the presence of dihydrotestosterone (DHT).</span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">Regulation of COVID-19 severity and fatality by sex hormones. Females have aromatase, the enzyme that converts androgen substrates into estrogen. On the other hand, males have steroid 5α reductase, the enzyme that is responsible for the conversion of testosterone into dihydrotestosterone (DHT). In case of males, DHT activates androgen receptor (AR) that binds to the androgen response element (ARE) present in the promoter of TMPRSS2 gene, leading to its transcription. This ultimately results into enhanced processing of viral spike protein for greater entry and spread of SARS-CoV-2 into host cells. On the other hand,in females, estrogen activates estrogen receptor (ER), which binds to the estrogen response element (ERE) present in the promoter of eNOS gene to drive its transcription and catalyze the formation of nitric oxide (NO) from L-arginine. This NO is involved in vasodilation as well as inhibition of viral replication. </span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif"><strong>NRP-1:</strong></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">For more information difference of NRP1 expression between male and female see <a href="https://www.proteinatlas.org/ENSG00000099250-NRP1/tissue"><span style="color:blue">https://www.proteinatlas.org/ENSG00000099250-NRP1/tissue</span></a><span style="color:blue">.</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">The expression of NRP1 protein in gastric cancer was not related to gender, age. The expression of NRP1 protein in gastric cancer is closely correlated to clinical stage, tumor size, TNM stage, differentiation, and lymph node metastasis (Cao et al., 2020).</span></span></p>
<p><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">SARS-CoV-2 Spike protein co-opts VEGF-A/Neuropilin-1 receptor signalling to induce analgesia had same results on both male and female rodents </span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">(Moutal et al., 2020)</span></span><span style="font-size:11.0pt"><span style="font-family:"Calibri",sans-serif">.</span></span></p>
UBERON:0000062organCL:0000000cellHighUnspecificHighAll life stagesHighLowModerateNot SpecifiedNot SpecifiedNot SpecifiedModerateHighHighModerate<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">BAUGHN, L. B., SHARMA, N., ELHAIK, E., SEKULIC, A., BRYCE, A. H. & FONSECA, R. 2020. Targeting TMPRSS2 in SARS-CoV-2 Infection. <em>Mayo Clin Proc,</em> 95<strong>,</strong> 1989-1999.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">BESTLE, D., HEINDL, M. R., LIMBURG, H., VAN LAM VAN, T., PILGRAM, O., MOULTON, H., STEIN, D. A., HARDES, K., EICKMANN, M., DOLNIK, O., ROHDE, C., KLENK, H.-D., GARTEN, W., STEINMETZER, T. & BÖTTCHER-FRIEBERTSHÄUSER, E. 2020a. TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells. <em>Life Science Alliance,</em> 3.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">BESTLE, D., HEINDL, M. R., LIMBURG, H., VAN LAM VAN, T., PILGRAM, O., MOULTON, H., STEIN, D. A., HARDES, K., EICKMANN, M., DOLNIK, O., ROHDE, C., KLENK, H. D., GARTEN, W., STEINMETZER, T. & BOTTCHER-FRIEBERTSHAUSER, E. 2020b. TMPRSS2 and furin are both essential for proteolytic activation of SARS-CoV-2 in human airway cells. <em>Life Sci Alliance,</em> 3.</span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">BUDZILOWICZ, C.J., WILCZYNSKI, S.P., AND WEISS, S.R. (1985). Three intergenic regions of coronavirus mouse hepatitis virus strain A59 genome RNA contain a common nucleotide sequence that is homologous to the 3' end of the viral mRNA leader sequence. <em>J Virol</em><em> 53</em>, 834-840.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">CANTUTI-CASTELVETRI, L., OJHA, R., PEDRO, L. D., DJANNATIAN, M., FRANZ, J., KUIVANEN, S., VAN DER MEER, F., KALLIO, K., KAYA, T., ANASTASINA, M., SMURA, T., LEVANOV, L., SZIROVICZA, L., TOBI, A., KALLIO-KOKKO, H., OSTERLUND, P., JOENSUU, M., MEUNIER, F. A., BUTCHER, S. J., WINKLER, M. S., MOLLENHAUER, B., HELENIUS, A., GOKCE, O., TEESALU, T., HEPOJOKI, J., VAPALAHTI, O., STADELMANN, C., BALISTRERI, G. & SIMONS, M. 2020. Neuropilin-1 facilitates SARS-CoV-2 cell entry and infectivity. <em>Science,</em> 370<strong>,</strong> 856-860.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">CAO, H., LI, Y., HUANG, L., BAI, B. & XU, Z. 2020. Clinicopathological Significance of Neuropilin 1 Expression in Gastric Cancer: A Meta-Analysis. <em>Dis Markers,</em> 2020<strong>,</strong> 4763492.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">CENTERS FOR DISEASE CONTROL AND PREVENTION, U. S. D. O. H. A. H. S. 2020. Covid-19 Science Update 2020.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">CHEN, L., LI, X., CHEN, M., FENG, Y. & XIONG, C. 2020. The ACE2 expression in human heart indicates new potential mechanism of heart injury among patients infected with SARS-CoV-2. <em>Cardiovasc Res,</em> 116<strong>,</strong> 1097-1100.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">DALY, J. L., SIMONETTI, B., KLEIN, K., CHEN, K. E., WILLIAMSON, M. K., ANTON-PLAGARO, C., SHOEMARK, D. K., SIMON-GRACIA, L., BAUER, M., HOLLANDI, R., GREBER, U. F., HORVATH, P., SESSIONS, R. B., HELENIUS, A., HISCOX, J. A., TEESALU, T., MATTHEWS, D. A., DAVIDSON, A. D., COLLINS, B. M., CULLEN, P. J. & YAMAUCHI, Y. 2020. Neuropilin-1 is a host factor for SARS-CoV-2 infection. <em>Science,</em> 370<strong>,</strong> 861-865.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">DAVIES, J., RANDEVA, H. S., CHATHA, K., HALL, M., SPANDIDOS, D. A., KARTERIS, E. & KYROU, I. 2020. Neuropilin1 as a new potential SARSCoV2 infection mediator implicated in the neurologic features and central nervous system involvement of COVID19. <em>Mol Med Rep,</em> 22<strong>,</strong> 4221-4226.</span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">DI, H., MCINTYRE, A.A., AND BRINTON, M.A. (2018). New insights about the regulation of Nidovirus subgenomic mRNA synthesis. <em>Virology</em><em> 517</em>, 38-43.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">GELFAND, M. V., HAGAN, N., TATA, A., OH, W. J., LACOSTE, B., KANG, K. T., KOPYCINSKA, J., BISCHOFF, J., WANG, J. H. & GU, C. 2014. Neuropilin-1 functions as a VEGFR2 co-receptor to guide developmental angiogenesis independent of ligand binding. <em>Elife,</em> 3<strong>,</strong> e03720.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">HARTENIAN, E., NANDAKUMAR, D., LARI, A., LY, M., TUCKER, J. M. & GLAUNSINGER, B. A. 2020. The molecular virology of coronaviruses. <em>J Biol Chem,</em> 295<strong>,</strong> 12910-12934.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">HASAN, A., PARAY, B. A., HUSSAIN, A., QADIR, F. A., ATTAR, F., AZIZ, F. M., SHARIFI, M., DERAKHSHANKHAH, H., RASTI, B., MEHRABI, M., SHAHPASAND, K., SABOURY, A. A. & FALAHATI, M. 2020. A review on the cleavage priming of the spike protein on coronavirus by angiotensin-converting enzyme-2 and furin. <em>J Biomol Struct Dyn</em><strong>,</strong> 1-9.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">HOFFMANN, M., KLEINE-WEBER, H. & POHLMANN, S. 2020a. A Multibasic Cleavage Site in the Spike Protein of SARS-CoV-2 Is Essential for Infection of Human Lung Cells. <em>Mol Cell,</em> 78<strong>,</strong> 779-784 e5.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">HOFFMANN, M., KLEINE-WEBER, H., SCHROEDER, S., KRUGER, N., HERRLER, T., ERICHSEN, S., SCHIERGENS, T. S., HERRLER, G., WU, N. H., NITSCHE, A., MULLER, M. A., DROSTEN, C. & POHLMANN, S. 2020b. SARS-CoV-2 Cell Entry Depends on ACE2 and TMPRSS2 and Is Blocked by a Clinically Proven Protease Inhibitor. <em>Cell,</em> 181<strong>,</strong> 271-280 e8.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">HUGGINS, D. J. 2020. Structural analysis of experimental drugs binding to the SARS-CoV-2 target TMPRSS2. <em>J Mol Graph Model,</em> 100<strong>,</strong> 107710.</span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">HUSTON, N.C., WAN, H., STRINE, M.S., DE CESARIS ARAUJO TAVARES, R., WILEN, C.B., AND PYLE, A.M. (2021). Comprehensive in vivo secondary structure of the SARS-CoV-2 genome reveals novel regulatory motifs and mechanisms. <em>Mol Cell</em><em> 81</em>, 584-598 e585.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">JING, Y., RUN-QIAN, L., HAO-RAN, W., HAO-RAN, C., YA-BIN, L., YANG, G. & FEI, C. 2020. Potential influence of COVID-19/ACE2 on the female reproductive system. <em>Mol Hum Reprod,</em> 26<strong>,</strong> 367-373.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">JOHNSON, B. A., XIE, X., KALVERAM, B., LOKUGAMAGE, K. G., MURUATO, A., ZOU, J., ZHANG, X., JUELICH, T., SMITH, J. K., ZHANG, L., BOPP, N., SCHINDEWOLF, C., VU, M., VANDERHEIDEN, A., SWETNAM, D., PLANTE, J. A., AGUILAR, P., PLANTE, K. S., LEE, B., WEAVER, S. C., SUTHAR, M. S., ROUTH, A. L., REN, P., KU, Z., AN, Z., DEBBINK, K., SHI, P. Y., FREIBERG, A. N. & MENACHERY, V. D. 2020. Furin Cleavage Site Is Key to SARS-CoV-2 Pathogenesis. <em>bioRxiv</em>.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">JU, B., ZHANG, Q., GE, J., WANG, R., SUN, J., GE, X., YU, J., SHAN, S., ZHOU, B., SONG, S., TANG, X., YU, J., LAN, J., YUAN, J., WANG, H., ZHAO, J., ZHANG, S., WANG, Y., SHI, X., LIU, L., ZHAO, J., WANG, X., ZHANG, Z. & ZHANG, L. 2020. Human neutralizing antibodies elicited by SARS-CoV-2 infection. <em>Nature,</em> 584<strong>,</strong> 115-119.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">JURASZEK, J., RUTTEN, L., BLOKLAND, S., BOUCHIER, P., VOORZAAT, R., RITSCHEL, T., BAKKERS, M. J. G., RENAULT, L. L. R. & LANGEDIJK, J. P. M. 2021. Stabilizing the closed SARS-CoV-2 spike trimer. <em>Nat Commun,</em> 12<strong>,</strong> 244.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">KANG, Y.-L., CHOU, Y.-Y., ROTHLAUF, P. W., LIU, Z., SOH, T. K., CURETON, D., CASE, J. B., CHEN, R. E., DIAMOND, M. S., WHELAN, S. P. J. & KIRCHHAUSEN, T. 2020. Inhibition of PIKfyve kinase prevents infection by Zaire ebolavirus and SARS-CoV-2. <em>bioRxiv</em><strong>,</strong> 2020.04.21.053058.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">KAWASE, M., SHIRATO, K., VAN DER HOEK, L., TAGUCHI, F. & MATSUYAMA, S. 2012. Simultaneous treatment of human bronchial epithelial cells with serine and cysteine protease inhibitors prevents severe acute respiratory syndrome coronavirus entry. <em>J Virol,</em> 86<strong>,</strong> 6537-45.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">KIM, D., LEE, J.Y., YANG, J.S., KIM, J.W., KIM, V.N., AND CHANG, H. (2020). The Architecture of SARS-CoV-2 Transcriptome. <em>Cell </em><em>181</em>, 914-921 e910.</span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">KLEIN, S., CORTESE, M., WINTER, S.L., WACHSMUTH-MELM, M., NEUFELDT, C.J., CERIKAN, B., STANIFER, M.L., BOULANT, S., BARTENSCHLAGER, R., AND CHLANDA, P. (2020). SARS-CoV-2 structure and replication characterized by in situ cryo-electron tomography.<em> Nat Commun</em> 11,<strong> </strong>5885.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LAM, S. D., BORDIN, N., WAMAN, V. P., SCHOLES, H. M., ASHFORD, P., SEN, N., VAN DORP, L., RAUER, C., DAWSON, N. L., PANG, C. S. M., ABBASIAN, M., SILLITOE, I., EDWARDS, S. J. L., FRATERNALI, F., LEES, J. G., SANTINI, J. M. & ORENGO, C. A. 2020. SARS-CoV-2 spike protein predicted to form complexes with host receptor protein orthologues from a broad range of mammals. <em>Sci Rep,</em> 10<strong>,</strong> 16471.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LAMERS, M. M., BEUMER, J., VAN DER VAART, J., KNOOPS, K., PUSCHHOF, J., BREUGEM, T. I., RAVELLI, R. B. G., PAUL VAN SCHAYCK, J., MYKYTYN, A. Z., DUIMEL, H. Q., VAN DONSELAAR, E., RIESEBOSCH, S., KUIJPERS, H. J. H., SCHIPPER, D., VAN DE WETERING, W. J., DE GRAAF, M., KOOPMANS, M., CUPPEN, E., PETERS, P. J., HAAGMANS, B. L. & CLEVERS, H. 2020. SARS-CoV-2 productively infects human gut enterocytes. <em>Science,</em> 369<strong>,</strong> 50-54.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LAN, J., GE, J., YU, J., SHAN, S., ZHOU, H., FAN, S., ZHANG, Q., SHI, X., WANG, Q., ZHANG, L. & WANG, X. 2020. Structure of the SARS-CoV-2 spike receptor-binding domain bound to the ACE2 receptor. <em>Nature,</em> 581<strong>,</strong> 215-220.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LETKO, M., MARZI, A. & MUNSTER, V. 2020. Functional assessment of cell entry and receptor usage for SARS-CoV-2 and other lineage B betacoronaviruses. <em>Nat Microbiol,</em> 5<strong>,</strong> 562-569.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LI, F. 2016. Structure, Function, and Evolution of Coronavirus Spike Proteins. <em>Annu Rev Virol,</em> 3<strong>,</strong> 237-261.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LIN, B., FERGUSON, C., WHITE, J. T., WANG, S., VESSELLA, R., TRUE, L. D., HOOD, L. & NELSON, P. S. 1999. Prostate-localized and androgen-regulated expression of the membrane-bound serine protease TMPRSS2. <em>Cancer Res,</em> 59<strong>,</strong> 4180-4.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LIU, L., WANG, P., NAIR, M. S., YU, J., RAPP, M., WANG, Q., LUO, Y., CHAN, J. F., SAHI, V., FIGUEROA, A., GUO, X. V., CERUTTI, G., BIMELA, J., GORMAN, J., ZHOU, T., CHEN, Z., YUEN, K. Y., KWONG, P. D., SODROSKI, J. G., YIN, M. T., SHENG, Z., HUANG, Y., SHAPIRO, L. & HO, D. D. 2020. Potent neutralizing antibodies against multiple epitopes on SARS-CoV-2 spike. <em>Nature,</em> 584<strong>,</strong> 450-456.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LU, Y. & MENG, Y. G. 2015. Quantitation of Circulating Neuropilin-1 in Human, Monkey, Mouse, and Rat Sera by ELISA. <em>Methods Mol Biol,</em> 1332<strong>,</strong> 39-48.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LUCAS, J. M., TRUE, L., HAWLEY, S., MATSUMURA, M., MORRISSEY, C., VESSELLA, R. & NELSON, P. S. 2008. The androgen-regulated type II serine protease TMPRSS2 is differentially expressed and mislocalized in prostate adenocarcinoma. <em>J Pathol,</em> 215<strong>,</strong> 118-25.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">LUKASSEN, S., CHUA, R. L., TREFZER, T., KAHN, N. C., SCHNEIDER, M. A., MULEY, T., WINTER, H., MEISTER, M., VEITH, C., BOOTS, A. W., HENNIG, B. P., KREUTER, M., CONRAD, C. & EILS, R. 2020. SARS-CoV-2 receptor ACE2 and TMPRSS2 are primarily expressed in bronchial transient secretory cells. <em>EMBO J,</em> 39<strong>,</strong> e105114.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MADJID, M., SAFAVI-NAEINI, P., SOLOMON, S. D. & VARDENY, O. 2020. Potential Effects of Coronaviruses on the Cardiovascular System: A Review. <em>JAMA Cardiol,</em> 5<strong>,</strong> 831-840.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MAYI, B. S., LEIBOWITZ, J. A., WOODS, A. T., AMMON, K. A., LIU, A. E. & RAJA, A. 2021. The role of Neuropilin-1 in COVID-19. <em>PLoS Pathog,</em> 17<strong>,</strong> e1009153.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MILLET, J. K. & WHITTAKER, G. R. 2018. Physiological and molecular triggers for SARS-CoV membrane fusion and entry into host cells. <em>Virology,</em> 517<strong>,</strong> 3-8.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MIRABELLI, C., WOTRING, J. W., ZHANG, C. J., MCCARTY, S. M., FURSMIDT, R., FRUM, T., KADAMBI, N. S., AMIN, A. T., O’MEARA, T. R., PRETTO, C. D., SPENCE, J. R., HUANG, J., ALYSANDRATOS, K. D., KOTTON, D. N., HANDELMAN, S. K., WOBUS, C. E., WEATHERWAX, K. J., MASHOUR, G. A., O’MEARA, M. J. & SEXTON, J. Z. 2020. Morphological Cell Profiling of SARS-CoV-2 Infection Identifies Drug Repurposing Candidates for COVID-19. <em>bioRxiv</em><strong>,</strong> 2020.05.27.117184.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MOUTAL, A., MARTIN, L. F., BOINON, L., GOMEZ, K., RAN, D., ZHOU, Y., STRATTON, H. J., CAI, S., LUO, S., GONZALEZ, K. B., PEREZ-MILLER, S., PATWARDHAN, A., IBRAHIM, M. M. & KHANNA, R. 2020. SARS-CoV-2 Spike protein co-opts VEGF-A/Neuropilin-1 receptor signaling to induce analgesia. <em>bioRxiv</em>.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MUHL, L., FOLESTAD, E. B., GLADH, H., WANG, Y., MOESSINGER, C., JAKOBSSON, L. & ERIKSSON, U. 2017. Neuropilin 1 binds PDGF-D and is a co-receptor in PDGF-D-PDGFRbeta signaling. <em>J Cell Sci,</em> 130<strong>,</strong> 1365-1378.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MUKHERJEE, S. & PAHAN, K. 2021. Is COVID-19 Gender-sensitive? <em>J Neuroimmune Pharmacol,</em> 16<strong>,</strong> 38-47.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">MURGOLO, N., THERIEN, A. G., HOWELL, B., KLEIN, D., KOEPLINGER, K., LIEBERMAN, L. A., ADAM, G. C., FLYNN, J., MCKENNA, P., SWAMINATHAN, G., HAZUDA, D. J. & OLSEN, D. B. 2021. SARS-CoV-2 tropism, entry, replication, and propagation: Considerations for drug discovery and development. <em>PLoS Pathog,</em> 17<strong>,</strong> e1009225.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">PAN, X. W., XU, D., ZHANG, H., ZHOU, W., WANG, L. H. & CUI, X. G. 2020. Identification of a potential mechanism of acute kidney injury during the COVID-19 outbreak: a study based on single-cell transcriptome analysis. <em>Intensive Care Med,</em> 46<strong>,</strong> 1114-1116.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">PEREZ-MILLER, S., PATEK, M., MOUTAL, A., CABEL, C. R., THORNE, C. A., CAMPOS, S. K. & KHANNA, R. 2020. In silico identification and validation of inhibitors of the interaction between neuropilin receptor 1 and SARS-CoV-2 Spike protein. <em>bioRxiv</em>.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">PREMKUMAR, L., SEGOVIA-CHUMBEZ, B., JADI, R., MARTINEZ, D. R., RAUT, R., MARKMANN, A., CORNABY, C., BARTELT, L., WEISS, S., PARK, Y., EDWARDS, C. E., WEIMER, E., SCHERER, E. M., ROUPHAEL, N., EDUPUGANTI, S., WEISKOPF, D., TSE, L. V., HOU, Y. J., MARGOLIS, D., SETTE, A., COLLINS, M. H., SCHMITZ, J., BARIC, R. S. & DE SILVA, A. M. 2020. The receptor binding domain of the viral spike protein is an immunodominant and highly specific target of antibodies in SARS-CoV-2 patients. <em>Sci Immunol,</em> 5.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">QIAO, Y., WANG, X. M., MANNAN, R., PITCHIAYA, S., ZHANG, Y., WOTRING, J. W., XIAO, L., ROBINSON, D. R., WU, Y. M., TIEN, J. C., CAO, X., SIMKO, S. A., APEL, I. J., BAWA, P., KREGEL, S., NARAYANAN, S. P., RASKIND, G., ELLISON, S. J., PAROLIA, A., ZELENKA-WANG, S., MCMURRY, L., SU, F., WANG, R., CHENG, Y., DELEKTA, A. D., MEI, Z., PRETTO, C. D., WANG, S., MEHRA, R., SEXTON, J. Z. & CHINNAIYAN, A. M. 2020. Targeting transcriptional regulation of SARS-CoV-2 entry factors ACE2 and TMPRSS2. <em>Proc Natl Acad Sci U S A</em>.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">RAHMAN, N., BASHARAT, Z., YOUSUF, M., CASTALDO, G., RASTRELLI, L. & KHAN, H. 2020. Virtual Screening of Natural Products against Type II Transmembrane Serine Protease (TMPRSS2), the Priming Agent of Coronavirus 2 (SARS-CoV-2). <em>Molecules,</em> 25.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">RAIMONDI, C., BRASH, J. T., FANTIN, A. & RUHRBERG, C. 2016. NRP1 function and targeting in neurovascular development and eye disease. <em>Prog Retin Eye Res,</em> 52<strong>,</strong> 64-83.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">RIVA, L., YUAN, S., YIN, X., MARTIN-SANCHO, L., MATSUNAGA, N., BURGSTALLER-MUEHLBACHER, S., PACHE, L., DE JESUS, P. P., HULL, M. V., CHANG, M., CHAN, J. F.-W., CAO, J., POON, V. K.-M., HERBERT, K., NGUYEN, T.-T., PU, Y., NGUYEN, C., RUBANOV, A., MARTINEZ-SOBRIDO, L., LIU, W.-C., MIORIN, L., WHITE, K. M., JOHNSON, J. R., BENNER, C., SUN, R., SCHULTZ, P. G., SU, A., GARCIA-SASTRE, A., CHATTERJEE, A. K., YUEN, K.-Y. & CHANDA, S. K. 2020. A Large-scale Drug Repositioning Survey for SARS-CoV-2 Antivirals. <em>bioRxiv</em><strong>,</strong> 2020.04.16.044016.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">ROBBIANI, D. F., GAEBLER, C., MUECKSCH, F., LORENZI, J. C. C., WANG, Z., CHO, A., AGUDELO, M., BARNES, C. O., GAZUMYAN, A., FINKIN, S., HAGGLOF, T., OLIVEIRA, T. Y., VIANT, C., HURLEY, A., HOFFMANN, H. H., MILLARD, K. G., KOST, R. G., CIPOLLA, M., GORDON, K., BIANCHINI, F., CHEN, S. T., RAMOS, V., PATEL, R., DIZON, J., SHIMELIOVICH, I., MENDOZA, P., HARTWEGER, H., NOGUEIRA, L., PACK, M., HOROWITZ, J., SCHMIDT, F., WEISBLUM, Y., MICHAILIDIS, E., ASHBROOK, A. W., WALTARI, E., PAK, J. E., HUEY-TUBMAN, K. E., KORANDA, N., HOFFMAN, P. R., WEST, A. P., JR., RICE, C. M., HATZIIOANNOU, T., BJORKMAN, P. J., BIENIASZ, P. D., CASKEY, M. & NUSSENZWEIG, M. C. 2020. Convergent antibody responses to SARS-CoV-2 in convalescent individuals. <em>Nature,</em> 584<strong>,</strong> 437-442.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">ROSSI, A., DEVERAUX, Q., TURK, B. & SALI, A. 2004. Comprehensive search for cysteine cathepsins in the human genome. <em>Biol Chem,</em> 385<strong>,</strong> 363-72.</span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SACCO, M.D., MA, C., LAGARIAS, P., GAO, A., TOWNSEND, J.A., MENG, X., DUBE, P., ZHANG, X., HU, Y., KITAMURA, N.<em>, et al.</em> (2020). Structure and inhibition of the SARS-CoV-2 main protease reveal strategy for developing dual inhibitors against M(pro) and cathepsin <em>L. Sci Adv</em><em> </em>6(50):eabe0751.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SHANG, J., WAN, Y., LUO, C., YE, G., GENG, Q., AUERBACH, A. & LI, F. 2020. Cell entry mechanisms of SARS-CoV-2. <em>Proc Natl Acad Sci U S A,</em> 117<strong>,</strong> 11727-11734.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SHIRATO, K., KAWASE, M. & MATSUYAMA, S. 2018. Wild-type human coronaviruses prefer cell-surface TMPRSS2 to endosomal cathepsins for cell entry. <em>Virology,</em> 517<strong>,</strong> 9-15.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SIMMONS, G., GOSALIA, D. N., RENNEKAMP, A. J., REEVES, J. D., DIAMOND, S. L. & BATES, P. 2005. Inhibitors of cathepsin L prevent severe acute respiratory syndrome coronavirus entry. <em>Proc Natl Acad Sci U S A,</em> 102<strong>,</strong> 11876-81.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SINGH, M., BANSAL, V. & FESCHOTTE, C. 2020. A single-cell RNA expression map of human coronavirus entry factors. <em>bioRxiv</em>.</span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SNIJDER, E.J., BREDENBEEK, P.J., DOBBE, J.C., THIEL, V., ZIEBUHR, J., POON, L.L.M., GUAN, Y., ROZANOV, M., SPAAN, W.J.M., AND GORBALENYA, A.E. (2003). Unique and Conserved Features of Genome and Proteome of SARS-coronavirus, an Early Split-off From the Coronavirus Group 2 Lineage. <em>Journal of Molecular Biology</em><em> 331</em>, 991-1004.</span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SNIJDER, E.J., LIMPENS, R., DE WILDE, A.H., DE JONG, A.W.M., ZEVENHOVEN-DOBBE, J.C., MAIER, H.J., FAAS, F., KOSTER, A.J., AND BARCENA, M. (2020). A unifying structural and functional model of the coronavirus replication organelle: Tracking down RNA synthesis. <em>PLoS Biol</em><em> 18</em>, e3000715.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">SUBRAMANIAN, A., VERNON, K. A., SLYPER, M., WALDMAN, J., LUECKEN, M. D., GOSIK, K., DUBINSKY, D., CUOCO, M. S., KELLER, K., PURNELL, J., NGUYEN, L., DIONNE, D., ROZENBLATT-ROSEN, O., WEINS, A., REGEV, A. & GREKA, A. 2020. RAAS blockade, kidney disease, and expression of ACE2, the entry receptor for SARS-CoV-2, in kidney epithelial and endothelial cells.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">UNIPROTKB - O14786 (NRP1_HUMAN) </span></span></p>
<p style="margin-left:40px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">V'KOVSKI, P., KRATZEL, A., STEINER, S., STALDER, H., AND THIEL, V. (2021). Coronavirus biology and replication: implications for SARS-CoV-2. <em>Nat Rev Microbiol</em>. 19,<strong> </strong>155–170.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">WALLS, A. C., TORTORICI, M. A., BOSCH, B. J., FRENZ, B., ROTTIER, P. J. M., DIMAIO, F., REY, F. A. & VEESLER, D. 2016. Cryo-electron microscopy structure of a coronavirus spike glycoprotein trimer. <em>Nature,</em> 531<strong>,</strong> 114-117.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">WANG, Y., LIU, M. & GAO, J. 2020. Enhanced receptor binding of SARS-CoV-2 through networks of hydrogen-bonding and hydrophobic interactions. <em>Proc Natl Acad Sci U S A,</em> 117<strong>,</strong> 13967-13974.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">YAMAMOTO, M., KISO, M., SAKAI-TAGAWA, Y., IWATSUKI-HORIMOTO, K., IMAI, M., TAKEDA, M., KINOSHITA, N., OHMAGARI, N., GOHDA, J., SEMBA, K., MATSUDA, Z., KAWAGUCHI, Y., KAWAOKA, Y. & INOUE, J. I. 2020. The Anticoagulant Nafamostat Potently Inhibits SARS-CoV-2 S Protein-Mediated Fusion in a Cell Fusion Assay System and Viral Infection In Vitro in a Cell-Type-Dependent Manner. <em>Viruses,</em> 12.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">YANG, N. & SHEN, H.-M. 2020. Targeting the Endocytic Pathway and Autophagy Process as a Novel Therapeutic Strategy in COVID-19. <em>International Journal of Biological Sciences,</em> 16<strong>,</strong> 1724-1731.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">YUAN, M., WU, N. C., ZHU, X., LEE, C. D., SO, R. T. Y., LV, H., MOK, C. K. P. & WILSON, I. A. 2020. A highly conserved cryptic epitope in the receptor binding domains of SARS-CoV-2 and SARS-CoV. <em>Science,</em> 368<strong>,</strong> 630-633.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">ZANG, R., GOMEZ CASTRO, M. F., MCCUNE, B. T., ZENG, Q., ROTHLAUF, P. W., SONNEK, N. M., LIU, Z., BRULOIS, K. F., WANG, X., GREENBERG, H. B., DIAMOND, M. S., CIORBA, M. A., WHELAN, S. P. J. & DING, S. 2020. TMPRSS2 and TMPRSS4 promote SARS-CoV-2 infection of human small intestinal enterocytes. <em>Sci Immunol,</em> 5.</span></span></p>
<p style="margin-left:48px"><span style="font-size:11pt"><span style="font-family:"Calibri",sans-serif">ZIEGLER, C. G. K., ALLON, S. J., NYQUIST, S. K., MBANO, I. M., MIAO, V. N., TZOUANAS, C. N., CAO, Y., YOUSIF, A. S., BALS, J., HAUSER, B. M., FELDMAN, J., MUUS, C., WADSWORTH, M. H., 2ND, KAZER, S. W., HUGHES, T. K., DORAN, B., GATTER, G. J., VUKOVIC, M., TALIAFERRO, F., MEAD, B. E., GUO, Z., WANG, J. P., GRAS, D., PLAISANT, M., ANSARI, M., ANGELIDIS, I., ADLER, H., SUCRE, J. M. S., TAYLOR, C. J., LIN, B., WAGHRAY, A., MITSIALIS, V., DWYER, D. F., BUCHHEIT, K. M., BOYCE, J. A., BARRETT, N. A., LAIDLAW, T. M., CARROLL, S. L., COLONNA, L., TKACHEV, V., PETERSON, C. W., YU, A., ZHENG, H. B., GIDEON, H. P., WINCHELL, C. G., LIN, P. L., BINGLE, C. D., SNAPPER, S. B., KROPSKI, J. A., THEIS, F. J., SCHILLER, H. B., ZARAGOSI, L. E., BARBRY, P., LESLIE, A., KIEM, H. P., FLYNN, J. L., FORTUNE, S. M., BERGER, B., FINBERG, R. W., KEAN, L. S., GARBER, M., SCHMIDT, A. G., LINGWOOD, D., SHALEK, A. K., ORDOVAS-MONTANES, J., LUNG-NETWORK@HUMANCELLATLAS.ORG, H. C. A. L. B. N. E. A. & NETWORK, H. C. A. L. B. 2020. SARS-CoV-2 Receptor ACE2 Is an Interferon-Stimulated Gene in Human Airway Epithelial Cells and Is Detected in Specific Cell Subsets across Tissues. <em>Cell,</em> 181<strong>,</strong> 1016-1035 e19.</span></span></p>
2020-03-01T10:29:312023-04-04T07:39:34blood brain barrier disruption BBB disruptionCellular2021-04-27T13:48:252021-04-27T13:48:25Thrombosis and Disseminated Intravascular CoagulationThrombosis and DICIndividual<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Thrombosis is defined as the formation or presence of a thrombus. Clotting within a blood vessel may cause infarction of tissues supplied by the vessel. Extreme aggravation of blood coagulation induces multiple thrombi in the microvasculature, which leads to consumption coagulopathy followed by disseminated intravascular coagulation (DIC).</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">DIC is a pathological syndrome resulting from the formation of thrombin, subsequent activation and consumption of coagulation proteins, and the production of fibrin thrombi. The initial pathologic events are thrombotic in nature resulting in thrombotic vascular occlusions. The initial clinical events are usually hemorrhagic resulting in oozing from mucosa and massive gastrointestinal blood loss. The occlusive events occur as a result of fibrin microthrombi or platelet microthrombi that obstruct the microcirculation of organs. This obstruction can result in organ hypoperfusion and ischemia, infarction, and necrosis. All organs are potentially vulnerable to the effects of thrombotic occlusions.</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">The renal effects of DIC are multifactorial and may be associated with hypovolemia or hypotension. If the hypotension is not corrected it may lead to renal failure due to acute tubular necrosis. Fibrin thrombi may also block glomerular capillaries causing ischemic, renal cortical necrosis (Colman, 1984).</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">The cerebral effects of DIC often result in nonspecific changes such as altered state of consciousness, convulsions, and coma. Major vascular occlusions, subarachnoid hemorrhage, multiple cortical and brain stem hemorrhages may occur following microvascular occlusions (Schwartzman RJ, 1982).</span></span></span></span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">The pulmonary effects of DIC may be caused by interstitial hemorrhage resulting in a clinical effect resembling acute respiratory distress syndrome (Schwartzman RJ,1973; Shahl RL, 1984).</span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Clinical laboratory tests are used to diagnose DIC.</span></span></span></span></span></p>
<p><strong><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#202124">Prothrombin time</span></span></span></strong><span style="background-color:white; font-family:Calibri,sans-serif; font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="color:#202124"> (<strong>PT</strong>) is a blood test that measures how long it takes blood to clot.</span></span></span><span style="background-color:white; font-family:Calibri,sans-serif; font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="color:#2e2e2e"> PT measures the time required for fibrin clot formation after the addition of tissue thromboplastin and calcium.</span></span></span><span style="background-color:white; font-family:Calibri,sans-serif; font-size:11pt"> </span><span style="background-color:white; font-family:Calibri,sans-serif; font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="color:#111111">The average time range for blood to clot is about 10 to 13 seconds.</span></span></span></p>
<p><strong><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Activated partial prothrombin time (APTT). </span></span></span></strong><span style="font-size:12.0pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"><span style="color:#333333">Platelet poor plasma [PPP] is incubated at 37°C then phospholipid (cephalin) and a contact activator (e.g. Kaolin, micronized silica, or ellagic acid) are added. This leads to the conversion of Factor XI [FXI] to FXIa. The remainder of the pathway is not activated as no calcium is present. The addition of calcium (pre-warmed to 37°C) initiates clotting. The APTT is the time taken from the addition of calcium to the formation of a fibrin clot.</span></span></span></span><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#333333"> The clotting time for the APTT lies between 27-35 seconds.</span></span></span></p>
<p> </p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Decreased fibrinogen concentrations</span></span></span></strong></span></span></p>
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<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:black">Diluted plasma is clotted with a high concentration of Thrombin. The tested plasma is diluted (usually 1:10 but this may vary if the Fibrinogen concentration is very low or very high) to minimize the effect of 'inhibitory substances' within the plasma e.g. heparin, elevated levels of FDPs. The use of a high concentration of Thrombin (typically 100 U/ml) ensures that the clotting times are independent of Thrombin concentration over a wide range of Fibrinogen levels.<br />
<br />
The test requires a reference plasma with a known Fibrinogen concentration and that has been calibrated against a known international reference standard. A calibration curve is constructed using this reference plasma by preparing a series of dilutions (1:5 –1:40) in the buffer to give a range of Fibrinogen concentrations. The clotting time of each of these dilutions is established (using duplicate samples) and the results (clotting time(s)/Fibrinogen concentration (g/L) are plotted on Log-Log graph paper. The 1:10 concentration is considered to be 100% i.e. normal. There should be a linear correlation between clotting times in the region of 10-50 sec.<br />
<br />
The test platelet-poor diluted plasma (diluted 1:10 in buffer) is incubated at 37°C, Thrombin [~100 U/mL] added (all pre-warmed to 37°C). The time taken for the clot to form is compared to the calibration curve and the Fibrinogen concentration deduced. Test samples whose clotting times fall out with the linear part of the calibration curve should be re-tested using different dilutions.<br />
<br />
Most laboratories use an automated method in which clot formation is deemed to have occurred when the optical density of the mixture has exceeded a certain threshold.</span></span></span></span></span></p>
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<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Platelet Measurements-</span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">A platelet count is the number of platelets a person has per microliter. The ideal platelet range is 150,000 – 400,000 per microliter in most healthy people.</span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Fibrinolysis measurements-</span></span></span></span></p>
<p><span style="font-size:12pt"><span style="background-color:white"><span style="font-family:"Times New Roman",serif"> d-dimer concentration <span style="font-size:9.0pt"><span style="font-family:"Verdana",sans-serif">ALERE TRIAGE® D-DIMER TEST</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">D-Dimer can be measured by a fluorescence immunoassay. To determine cross-linked fibrin degradation products containing D-dimer in EDTA anticoagulated whole blood and plasma specimens. The test is used as an aid in the assessment and evaluation of patients suspected of having disseminated intravascular coagulation or thromboembolic events including pulmonary embolism</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="background-color:white"><span style="font-family:Calibri,sans-serif"><strong><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Procedure: </span></span></strong></span></span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif">Commercially available kits are available to measure d-dimer in whole blood or plasma. The kits contain all the reagents necessary for the quantification of cross-linked fibrin degradation products containing D-dimer in EDTA anticoagulated whole blood or plasma specimens.</span></span></p>
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<p><span style="font-family:Times New Roman,Times,serif">Homo sapiens</span></p>
UBERON:0000178bloodNot SpecifiedUnspecificNot SpecifiedAll life stagesModerate<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Hemostasis and Thrombosis Basic Principles and Clinical Practices Robert W Colman, Jack Hirsh, Victor J. Marder, Edwin W. Salzman (ed) Philadelphia, 1994.</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Schwartzman RJ, Hill JB: Neurologic complications of DIC. Neurology 32:791, 1982</span></span></span></span></span></p>
<p><span style="font-size:11pt"><span style="font-family:Calibri,sans-serif"><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Robboy SJ, Minna JD, Colman RW et.al. Pulmonary hemorrhage syndrome as a manifestation of DIC: Analysis of 10 cases. Chest 63:718, 1973.</span></span></span></span></span></p>
<p><span style="font-size:12.0pt"><span style="font-family:"Times New Roman",serif"><span style="color:#444444">Stahl RL, Javid JP, Lackner H: Unrecognized pulmonary embolism presenting as DIC. SM J Med 76:772, 1984.</span></span></span></p>
2021-03-25T19:52:452022-11-25T01:38:01Cerebrovascular disease (stroke)strokeOrgan<p><span style="color:#000000"><span style="font-family:Calibri">A stroke or cerebrovascular disease (CVD) is the result of an acute disruption of the cerebral perfusion or vasculature (Khaku and Tadi, 2020). The majority of strokes are of ischemic origin and only a small percent of hemorrhagic origin. Ischemic stroke can be further characterised into embolic, thrombotic, and lacunar. </span></span></p>
<p><span style="color:#000000"><span style="font-family:Calibri">Lacunar strokes result from disruption of the small penetrating branches of the middle cerebral artery, vertebral or basilar artery or the lenticulostriate vessels. Most common causes of lacunar strokes include microemboli and fibrinoid necrosis secondary to vasculitis. </span></span></p>
<p><span style="color:#000000"><span style="font-family:Calibri">Hemorrhagic etiologies can be from aneurysm rupture, arteriovenous malformations, venous angiomas, bleeding due to drugs.</span></span></p>
<p><span style="color:#000000"><span style="font-family:Calibri">In general, the common risk factors for stroke include hypertension, diabetes, smoking, obesity, atrial fibrillation, and drug use. </span></span></p>
<p><span style="color:#000000"><span style="font-family:Calibri">Patients with CVD can show any of the following symptoms </span></span></p>
<ul>
<li><span style="font-size:11pt">Disturbance of consciousness (e.g. not alert and responsive, arouses to noxious stimuli, comatose…)</span></li>
<li><span style="font-size:11pt">Language difficulties in fluency, naming, comprehension and repetition </span></li>
<li><span style="font-size:11pt">Dysarthria (slurring) </span></li>
<li><span style="font-size:11pt">Motor deficits (e.g. subtle arm weakness)</span></li>
<li><span style="font-size:11pt">Visual field deficits</span></li>
<li><span style="font-size:11pt">Eye movement abnormalities </span></li>
<li><span style="font-size:11pt">Facial paralysis </span></li>
<li><span style="font-size:11pt">Ataxia </span></li>
<li><span style="font-size:11pt">Nausea, </span></li>
<li><span style="font-size:11pt">Vomiting </span></li>
<li><span style="font-size:11pt">Vertigo</span></li>
</ul>
<p><span style="color:#000000"><span style="font-family:Calibri">Based on the symptoms, the localisation of the stroke in brain areas can be diagnosed and also support the categorisation to the following stroke syndromes: a) Anterior Cerebral Artery (ACA) Infarction, b) Middle Cerebral Artery (MCA) Infarction, c) Posterior Cerebral Artery (PCA) Infarction and c) Cerebellar Infarction.</span></span></p>
<p><span style="color:#000000"><span style="font-family:Calibri">It is detected by clinical examination stat non-contrasted head computed tomogram (CT) or a combination of head CT, CT Angiography, and perfusion imaging (Khaku and Tadi, 2020).</span></span></p>
Not SpecifiedMaleNot SpecifiedFemaleNot SpecifiedAdultNot SpecifiedOld AgeNot Specified<p><span style="color:#000000"><span style="font-family:Calibri">Khaku AS, Tadi P. (2020) Cerebrovascular Disease. In: StatPearls. Treasure Island (FL): StatPearls Publishing; 2020 Jan-. Available from: https://www.ncbi.nlm.nih.gov/books/NBK430927/</span></span></p>
2021-04-27T13:50:082022-03-30T11:30:53Binding of Sars-CoV-2 spike protein to ACE 2 receptors expressed on pericytes leads to disseminated intravascular coagulation resulting in cerebrovascular disease (stroke)Sars-CoV-2 causes strokeUnder development: Not open for comment. Do not citeUnder DevelopmentIncluded in OECD Work Plan1.96<p><span style="font-size:9.0pt"><span style="font-family:"Times New Roman",serif">Receptor recognition is an essential determinant of molecular level in this AOP. ACE2 was reported as an entry receptor for SARS-CoV-2. The viral entry process is mediated by the envelope-embedded surface-located spike (S) glycoprotein. Jun Lan and Walls, A.C et al (Nature 581, 215–220; Cell 180, 281–292) demonstrated a critical initial step of infection at the molecular level from the interaction of ACE2 and S protein. ACE2 has shown that receptor binding affinity to S protein is nM range. To elucidate the interaction between the SARS-CoV-2 RBD and ACE2 at a higher resolution, they also determined the structure of the SARS-CoV-2 RBD–ACE2 complex using X-ray crystallography.</span></span> <span style="font-size:9.0pt"><span style="font-family:"Times New Roman",serif">The expression and distribution of the ACE2 in human body may indicate the potential infection of SARS-CoV-2. Through the developed single-cell RNA sequencing (scRNA-Seq) technique and single-cell transcriptomes based on the public database, researchers analyzed the ACE2 RNA expression profile at single-cell resolution. High ACE2 expression was identified in type II alveolar cells (Zou, X. et al.</span></span> <span style="font-size:9.0pt"><span style="font-family:"Times New Roman",serif">Front. Med.2020)</span></span></p>
<p><span style="font-size:12px"><span style="font-family:"Times New Roman",serif">SARS-CoV-2 belongs to the Coronaviridae family, which includes evolutionary related enveloped (+) strand RNA viruses of vertebrates, such as seasonal common coronaviruses, SARS-CoV and CoV-NL63, SARS-CoV (Kim Young Jun et al)</span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Human viruses strains</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Genus</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Major cell receptor</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">First report</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Animal reservoir</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Intermediate host</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Pathology</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Diagnostic test</span></span></span></span></p>
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<p style="text-align:center"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:white">Evidence</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">HCoV-NL63</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Alphacoronavirus</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">ACE2</span></span></span></span></p>
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<p style="text-align:right"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">2004</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Bat</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Unknown</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Mild respiratory tract illness</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">RT-PCR, IF, ELISA, WB</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Strong</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">SARS-CoV</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Betacoronavirus</span></span></span></span></p>
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<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">ACE2</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:172px">
<p style="text-align:right"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">2003</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:172px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Bat</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:204px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Pangolin</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:340px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Severe acute respiratory syndrome</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:252px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">RT-PCR, IF, ELISA, WB</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:182px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Strong</span></span></span></span></p>
</td>
</tr>
<tr>
<td style="border-bottom:1px solid black; border-left:1px solid black; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:206px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">SARS-CoV-2</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:172px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Betacoronavirus</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:172px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">ACE2</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:172px">
<p style="text-align:right"><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">2020</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:172px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Bat</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:204px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Pangolin</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:340px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Severe acute respiratory syndrome</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:252px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">RT-PCR, IF, ELISA, WB</span></span></span></span></p>
</td>
<td style="border-bottom:1px solid black; border-left:none; border-right:1px solid black; border-top:none; height:19px; vertical-align:bottom; width:182px">
<p><span style="font-size:12pt"><span style="font-family:"Times New Roman",serif"><span style="font-size:9.0pt"><span style="color:black">Strong</span></span></span></span></p>
</td>
</tr>
</tbody>
</table>
2021-04-27T13:41:532023-04-29T13:02:19