Distribution of ACE2, CD147, CD26, and other SARS-CoV-2 associated molecules in tissues and immune cells in health and in asthma, COPD, obesity, hypertension, and COVID-19 risk factors

Abstract

Background: Morbidity and mortality from COVID-19 caused by novel coronavirus SARS-CoV-2 is accelerating worldwide, and novel clinical presentations of COVID-19 are often reported. The range of human cells and tissues targeted by SARS-CoV-2, its potential receptors and associated regulating factors are still largely unknown. The aim of our study was to analyze the expression of known and potential SARS-CoV-2 receptors and related molecules in the extensive collection of primary human cells and tissues from healthy subjects of different age and from patients with risk factors and known comorbidities of COVID-19.

Methods: We performed RNA sequencing and explored available RNA-Seq databases to study gene expression and co-expression of ACE2, CD147 (BSG), and CD26 (DPP4) and their direct and indirect molecular partners in primary human bronchial epithelial cells, bronchial and skin biopsies, bronchoalveolar lavage fluid, whole blood, peripheral blood mononuclear cells (PBMCs), monocytes, neutrophils, DCs, NK cells, ILC1, ILC2, ILC3, CD4+ and CD8+ T cells, B cells, and plasmablasts. We analyzed the material from healthy children and adults, and from adults in relation to their disease or COVID-19 risk factor status.

Results: ACE2 and TMPRSS2 were coexpressed at the epithelial sites of the lung and skin, whereas CD147 (BSG), cyclophilins (PPIA andPPIB), CD26 (DPP4), and related molecules were expressed in both epithelium and in immune cells. We also observed a distinct age-related expression profile of these genes in the PBMCs and T cells from healthy children and adults. Asthma, COPD, hypertension, smoking, obesity, and male gender status generally led to the higher expression of ACE2- and CD147-related genes in the bronchial biopsy, BAL, or blood. Additionally, CD147-related genes correlated positively with age and BMI. Interestingly, we also observed higher expression of CD147-related genes in the lesional skin of patients with atopic dermatitis.

Conclusions: Our data suggest different receptor repertoire potentially involved in the SARS-CoV-2 infection at the epithelial barriers and in the immune cells. Altered expression of these receptors related to age, gender, obesity and smoking, as well as with the disease status, might contribute to COVID-19 morbidity and severity patterns.

Keywords: COPD; COVID-19; COVID-19 children; SARS receptor; asthma; hypertension; obesity.

Conflict of interest statement

Dr Radzikowska has nothing to disclose. Dr Ding has nothing to disclose. Dr Tan has nothing to disclose. Dr Zhakparov has nothing to disclose. Dr Peng has nothing to disclose. Dr Wawrzyniak has nothing to disclose. Dr Wang has nothing to disclose. Dr LI has nothing to disclose. Dr Morita has nothing to disclose. Dr Altunbulakli has nothing to disclose. Dr Reiger reports personal fees from Bencard, Germany, personal fees from Roche‐Posay, Germany, personal fees from Galderma, Germany, personal fees from Sebapharma, Germany, grants from CLR, Germany, outside the submitted work. Dr Neumann has nothing to disclose. Dr Lunjani has nothing to disclose. Dr Traidl‐Hoffmann reports grants and personal fees from Töpfer, personal fees from Sanofi, personal fees from Novartis, grants and personal fees from Sebapharma, grants and personal fees from Danone Nutricia, personal fees from La Roche Posay, personal fees from Lilly Pharma, personal fees from Mylan, outside the submitted work. Dr Nadeau reports grants and other from NIAID, other from Novartis, personal fees and other from Regeneron, grants and other from FARE, grants from EAT, other from Sanofi, other from Astellas, other from Nestle, other from BeforeBrands, other from Alladapt, other from ForTra, other from Genentech, other from AImmune Therapeutics, other from DBV Technologies, personal fees from Astrazeneca, personal fees from ImmuneWorks, personal fees from Cour Pharmaceuticals, grants from Allergenis, grants from Ukko Pharma, other from AnaptysBio, other from Adare Pharmaceuticals, other from Stallergenes‐Greer, other from NHLBI, other from NIEHS, other from EPA, other from WAO Center of Excellence, other from Iggenix, other from Probio, other from Vedanta, other from Centecor, other from Seed, from Immune Tolerance Network, from NIH, outside the submitted work. In addition, Dr Nadeau has a patent Inhibition of Allergic Reaction to Peanut Allergen using an IL‐33 Inhibitor pending, a patent Special Oral Formula for Decreasing Food Allergy Risk and Treatment for Food Allergy pending, a patent Basophil Activation Based Diagnostic Allergy Test pending, a patent Granulocyte‐based methods for detecting and monitoring immune system disorders pending, a patent Methods and Assays for Detecting and Quantifying Pure Subpopulations of White Blood Cells in Immune System Disorders pending, a patent Mixed Allergen Compositions and Methods for Using the Same pending, and a patent Microfluidic Device and Diagnostic Methods for Allergy Testing Based on Detection of Basophil Activation pending. Dr O'Mahony reports personal fees from AHL, grants from GSK, outside the submitted work. Dr Akdis reports grants from Allergopharma, Idorsia, Swiss National Science Foundation, Christine Kühne‐Center for Allergy Research and Education, European Commission's Horison's 2020 Framework Programme, Cure, Novartis Research Institutes, Astra Zeneca, Scibase, Glakso Smith‐Kline and other from Sanofi & Regeneron. Dr Sokolowska reports grants from SNSF, grants from GSK, outside the submitted work.

© 2020 EAACI and John Wiley and Sons A/S. Published by John Wiley and Sons Ltd.

Figures

Figure 1

ACE2 and TMPRSS2 are expressed at the barrier sites, whereas CD147, cyclophilins, CD26, and their interaction partners are present in immune cells and in epithelium. Expression of A) ACE2, B) TMPRSS2, C) BSG, D) PPIA, E) PPIB, F) S100A9, G) SLC16A7, H) SLC2A1, I) CD44, J) ITGB1, K) NFATC3, L) DPP4 genes in in vitro air liquid interface (ALI)—differentiated human primary bronchial epithelial cells (n = 10) and in ex vivo human primary bronchial biopsies (n = 5), bronchoalveolar fluid cells (n = 5), whole blood (n = 5), neutrophils (n = 4), classical monocytes (n = 4), plasmacytoid dendritic cells (n = 4), group 1, 2, and 3 innate lymphoid cells (n = 6 per group), natural‐killer cells (n = 4), naïve CD4+ T cells (n = 4), terminal effector CD4+ T cells (n = 2), naïve CD8+ T cells (n = 4), effector memory CD8+ T cells (n = 4), naïve B cells (n = 4), plasmablasts (n = 4), and skin biopsies (n = 6) from healthy adults. Data obtained from in vitro approaches are highlighted in red. Names of the proteins encoded by analyzed genes are stated in the brackets. HBECS, human bronchial epithelial cells; Bronch. biop., bronchial biopsy; BAL, bronchoalveolar fluid cells; Class. monocytes, classical monocytes; pDCs, plasmacytoid dendritic cells; ILC1, group 1 innate lymphoid cells; ILC2, group 2 innate lymphoid cells; ILC3, group 3 innate lymphoid cells; NK, natural killer cells; naïve CD4+, naïve CD4+ T cells; term. eff. CD4+, terminal effector CD4+ T cells; naïve CD8+, naïve CD8+ T cells; eff. mem. CD8+, effector memory CD8+ T cells

Figure 2

Distinct expression profile of ACE2‐, CD147‐, and CD26‐related genes in PBMCs and T cells of children and adults. A) Expression of ACE2‐, CD147‐, NFAT‐, and CD‐26‐related genes in the human PBMCs in healthy children aged 5‐17 months (n = 21), 12‐36 month (n = 14), 4‐16 years (n = 16) and healthy adults aged 16‐67 years (n = 19). B) Expression of ACE2‐, CD147‐, NFAT‐, and CD‐26‐related genes in the human naïve CD4+ T cells from 12 months old healthy children (n = 18) and 20‐35 years old healthy adults (n = 4). All heatmaps display normalized gene expression across the groups (rows normalization). Color‐coding represents gene families related to ACE2 (orange), CD147 (green), NF‐ATs (purple) and CD26 (yellow). MO, months old; YO, years old; PBMCs, peripheral blood mononuclear cells

Figure 3

Asthma, COPD, hypertension, smoking, obesity and gender are associated with differential expression of ACE2‐, CD147‐, and CD26‐related genes in immune cells and tissues. A) Differential expression of ACE2, TMPRSS2, BSG, SLC2A1, CD44, and ITGA3 genes in in vitro air liquid interface (ALI)‐differentiated human primary bronchial epithelial cells from non‐diseased controls (n = 5), asthma (n = 6) and COPD (n = 5) patients. B) Differential expression of ACE2, BSG, SLC7A5, ITGA3, ITGA6 genes in bronchial biopsies from non‐diseased controls (n = 16), patients with asthma (n = 22) and COPD (n = 3), or in comparison of smokers (n = 21) with non‐smoking individuals (n = 19). C) Differential expression of BSG, PPIA, S100A9, CD44, SLC16A7, SLC16A3, ITGA3, NFATC1, NFATC2 genes in the bronchoalveolar fluid (BAL) from the control individuals (n = 16), patients with asthma (n = 22) and COPD (n = 2), or in comparison of hypertensive (n = 9) with normotensive (n = 31) individuals; smokers (n = 20) with non‐smokers (n = 19); obese (n = 21) with non‐obese (n = 19); and males (n = 26) with females (n = 14). D) Differential expression of BSG, PPIA, S100A9, CD44, SLC16A7, ITGA6, NFATC2, LGALS3 and NOD2 genes in the whole blood of non‐diseased controls (n = 17), patients with asthma (n = 21) and COPD (n = 3), or in comparison of hypertensive (n = 9) with normotensive (n = 32) individuals; smokers (n = 21), with non‐smokers (n = 19); obese (n = 21) with non‐obese individuals (n = 20); and males (n = 27) with females (n = 14). Names of the proteins encoded by analyzed genes are stated in the brackets. *P < .05, **P < .01, ***P < .001, ****P < .0001. HBECS, human bronchial epithelial cells; Bronch. biop., bronchial biopsy; BAL, bronchoalveolar fluid cells; COPD, chronic obstructive pulmonary disease

Figure 4

Expression of certain CD147‐related genes correlates with BMI and age in the BAL and blood. Correlation of A) SLC16A3 expression and BMI, B) ITGA3 expression and BMI, C) LGALS3 expression and BMI, and D) CD44 expression and age in the bronchoalveolar fluid (BAL). Correlation of E) BSG expression and BMI, F) PPIA expression and BMI, G) S100A9 expression and BMI, H) CD44 expression and BMI, I) LGALS3 expression and BMI, J) SLC16A3 and age in the whole blood. Spearman correlation coefficient (r) was calculated, with the threshold of significance set to P = .05. Names of the proteins encoded by analyzed genes are stated in the brackets. BAL, bronchoalveolar fluid cells; BMI, body mass index

Figure 5

Unique expression profile of ACE2‐ and CD147‐related genes in lesional skin in patients with atopic dermatitis. Expression of ACE2, TMPRSS2, BSG, PPIA, PPIB, S100A9, CD44, SLC16A1, SLC16A3, SLC7A5, SLC3A2, SLC2A1, ITGA3, ITGA6, NFATC3, JUP, NME1, NOD2, SDC1, andDPP4 genes in the skin of healthy controls (n = 6) and in the lesional (n = 11), and non‐lesional (n = 11) skin of atopic dermatitis patients. Names of the proteins encoded by analyzed genes are stated in the margins. *P < .05, **P < .01, ***P < .001, ****P < .0001

Figure 6

Summary of the tissue and cellular expression, and models of A) ACE2, B) CD147, C) CD26, and their interaction partners. Please refer to the text for further details. CypA, Cyclophilin A; CypB, Cyclophilin B; HA, Hyaluronic acid; Gal‐3, Galectin 3; MCTs, Monocarboxylate transporters. Figure created with BioRender.com