key: cord-0255186-sepl0vbh authors: Keskus, Ayse Gokce; Tombaz, Melike; Arici, Burcin I.; Dincaslan, Fatma B.; Nabi, Afshan; Shehwana, Huma; Konu, Ozlen title: ace2 expression is higher in intestines and liver while being tightly regulated in development and disease in zebrafish date: 2020-12-24 journal: bioRxiv DOI: 10.1101/2020.12.24.424209 sha: 1be9c8e8f293d46d1e845e1374ee668beaa06326 doc_id: 255186 cord_uid: sepl0vbh Human Angiotensin I Converting Enzyme 2 (ACE2) that acts as a receptor for SARS-CoV-2 entry is highly expressed in human type II pneumocytes and enterocytes and similarly in other mammals and zebrafish (Danio rerio). The zebrafish genome has a highly conserved, one-to-one ortholog of ACE2, i.e., ace2, whose expression profile however has not yet been studied during development or in pathologies relevant to COVID-19. Herein, we identified significant development-, tissue- and gender-specific modulations in ace2 expression based on meta-analysis of zebrafish Affymetrix transcriptomics datasets (ndatasets=107, GPL1319 in GEO database). Co-expression network analysis of ace2 revealed distinct positively correlated (carboxypeptidase activity and fibrin clot formation), and negatively correlated (cilia biogenesis/transport and chromatin modifications) STRING network modules. Using additional transcriptomics datasets, we showed zebrafish embryos before 3 days post fertilization (dpf) exhibited low levels of ace2 that increased significantly until 4 dpf implicating a role for ace2 in organogenesis. Re-analysis of RNA-seq datasets from zebrafish adult tissues demonstrated ace2 was expressed highly in intestines, variably in liver, and at lower levels in other organs. In addition, zebrafish females and males showed significant dimorphism in their age-dependent expression of ace2, and between ovary and testis where the latter had higher levels. Moreover, we demonstrated ace2 expression was significantly modulated under different physiological and pathological conditions associated with development, diet, infection, and inflammation. Our findings implicate a novel translational role for zebrafish ace2 in differentiation and pathologies predominantly found in intestines and liver, in which the effects of SARS-CoV-2 could be detrimental. 117 also was calculated for each gene based on log2 transformed expression values for larval 118 development, using GSE24616 (2 dpf-8 dpf) and GSE38575 (EtOH treated samples only; 2dpf -119 7dpf) datasets; for intestine, using GSE35889 and GSE12189 (GFP+ samples only); and for liver, 120 using GSE74244 (liver samples only) and GSE100583, separately (Table S2 ). race2 value with 121 minimum p value was selected for multiple probe sets with the same Ensembl ID. Ensembl IDs 122 along with race2 values were used for identifying local network clusters using STRING enrichment 123 analysis with values for each of the above datasets (Szklarczyk et al., 2015) . Comparative Network Enrichment Analysis (CNEA) of Sting local networks were performed separately for development, intestine and liver datasets. Enriched local networks with the largest gene set was 126 selected in the case of multiple local networks present with the same name. Significant networks 127 were clustered and visualized in Cytoscape (Shannon et al., 2003) ; where each node represented 128 a local network, colored according to STRING enrichment score (the mean enrichment score in 129 the case of liver and intestine comparisons), having edge weights determined by the number of 130 shared genes with other connected networks. Selected local networks were visualized in detail, where each node on the PPI network obtained from STRING local network data referred to a gene 132 and was colored according to race2. In addition, GO term enrichment analysis was performed for 133 liver datasets using clusterprofiler package in R (Yu et al., 2012) . S1C ). In addition, we found ace2 was lowly expressed in embryogenesis 161 and started increasing at 3 dpf and thereafter until 4 dpf, after which was steadily expressed at 162 high levels (Figs. 2A, S1C). Analysis of yet another dataset (GSE38575) supported this finding of 163 an increase in ace2 expression after 3 dpf (Fig. 2B) . STRING enrichment scores based on race2 164 between ace2 and every other gene obtained separately for GSE24616 (2dpf -8dpf) and To support the role of ace2 in intestinal function and differentiation we next investigated the tissue 207 specificity of ace2 expression across multiple datasets we complied from RNA-seq experiments 208 ( Fig. 3B ; Table S1 ). Intestine and liver exhibited relatively high ace2 expression in comparison 209 with brain, gills, and kidney in that descending order (Fig. 3B ). Accordingly, zebrafish ace2 In addition to altered lipid metabolism, interferon pathway modulation (ISG15 antiviral mechanism 343 network) has arisen as a discriminator of low and high ace2 expressing liver in zebrafish. Interestingly, studies with SARS-CoV-2 reported ACE2 as an interferon stimulated gene (ISG) in Although the mortality risk of COVID-19 is relatively higher in males than females, there is no In conclusion, COVID-19 is a multiorgan disease affecting lungs, kidney, intestines, and liver beta-glucan from barley and its 434 lipid-lowering capacity: a meta-analysis of randomized, controlled trials Nonalcoholic fatty liver disease is 601 associated with COVID-19 severity independently of metabolic syndrome: a retrospective case-602 control study Epigenetic control 605 of intestinal barrier function and inflammation in zebrafish Differential expression analysis of 608 multifactor RNA-Seq experiments with respect to biological variation Possible role of L-form switching in 612 recurrent urinary tract infection The benefits of Vitamin D in the COVID-19 pandemic: 615 biochemical and immunological mechanisms Short bowel mucosal morphology, proliferation and inflammation at 618 first and repeat 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