Fig 2: ACE2 repression by EMT in lung cancer cells. (A) ACE2, EMT score, EMT group (epithelial or mesenchymal), CDH1, and ZEB1 expression with miR200 family miRNAs in NSCLC, SCLC, and HNSCC cell lines. (B) Venn diagrams revealing miRNAs correlated with ACE2 expression in both NSCLC and SCLC cell lines (upper) and TCGA LUAD and HNSC tumor biopsies (lower). RNAseq data are revealed as normalized relative gene expression. (C, D) Expression of (C) ACE2 and (D) ZEB1 after induction of EMT by TGF-β treatment in H358 and HCC827 NSCLC cell lines. (E) Expression of ACE2 and ZEB1 in NSCLC and HNSCC cell lines. (F) Expression of Ace2 in 393P murine lung adenocarcinoma cells with Zeb1 overexpression. Cell surface expression of (G) ACE2 and (H) Vimentin in HCC827 cells with constitutive overexpression of ZEB1. (I) ACE2, TMPRSS2, and GLUL expression in NSCLC cell lines with acquired resistance to EGFR TKIs that occurred through EMT but not when resistance occurred through EGFR T790M resistance mutations. RNAseq data are revealed as normalized relative gene expression. EMT, epithelial-to-mesenchymal transition; FC, fold change; HNSC, head and neck squamous cell (tumors); HNSCC, head and neck squamous cell carcinoma (cell lines); LUAD, lung adenocarcinoma; miRNA, microRNA; OE, overexpressing; Par, parental; RNAseq, RNA sequencing; TCGA, The Cancer Genome Atlas; TGF-β, transforming growth factor-β; TKI, tyrosine kinase inhibitor; Vect, vector.

Fig 3: EMT modulation for treatment of SARS-CoV-2. a, ZEB1 expression following 24h of 0.5 µM bemcentinib (Bem) treatment in mesenchymal Calu-1 cells or following over-expression of Zeb1 in 393P murine cells, as well as vimentin, E-cadherin and ACE2 in Calu-1 cells. b, ACE2 levels following 24h and 72 of 1 µM Bem treatment in HCC827 cells with constitutive overexpression of ZEB1. c, ACE2, GLUL, and Vimentin levels in a subset of NSCLC cell lines following 48h treatment with 100 µM hydroxychloroquine. d, Working model demonstrating aerodigestive/respiratory cells with ACE2 expression are epithelial and metabolically primed for replication [left] while infected cells demonstrate a shift to a mesenchymal phenotype with reversal of glutamine production, a leaky cell membrane and increased ZEB1 to induce EMT. [right]. Bem = bemcentenib.

Fig 4: ACE2 repression by EMT in lung cancer cells. a, ACE2, EMT score, EMT group (epithelial or mesenchymal), CDH1, and ZEB1 expression with miR200 family miRNAs in NSCLC, SCLC and HNSCC cell lines. b, Venn diagrams demonstrating miRNAs correlated with ACE2 expression in both NSCLC and SCLC cell lines (upper) and TCGA LUAD and HNSC tumor biopsies (lower). RNAseq data is shown as normalized relative gene expression. c,d,, Expression of ACE2 (c) and ZEB1(d) following induction of EMT by TGFß treatment in H358 and HCC827 NSCLC cell lines. e, Expression of ACE2 and ZEB1 in NSCLC and HNSCC cell lines. f, Expression of Ace2 in 393P murine lung adenocarcinoma cells with Zeb1 overexpression. g,h, Cell surface expression of ACE2 (g) and Vimentin (h) in HCC827 cells with constitutive overexpression of ZEB1. i, ACE2, TMPRSS2 and GLUL expression in NSCLC cell lines with acquired resistance to EGFR TKIs that occurred through EMT but not when resistance occurred through EGFR T790M resistance mutations. RNAseq data is shown as normalized relative gene expression.

Fig 5: EMT modulation for treatment of SARS-CoV-2. (A) ZEB1 expression after 24 h of 0.5 μM Bem treatment in mesenchymal Calu-1 cells or after overexpression of Zeb1 in 393P murine cells, including vimentin, E-cadherin, and ACE2 in Calu-1 cells. (B) ACE2 levels after 24 h and 72 of 1 μM Bem treatment in HCC827 cells with constitutive overexpression of ZEB1. (C) ACE2, GLUL, and Vimentin levels in a subset of NSCLC cell lines after 48 h treatment with 100 μM hydroxychloroquine. (D) Working model revealing aerodigestive/respiratory cells with ACE2 expression are epithelial and metabolically primed for replication (left), whereas infected cells reveal a shift to a mesenchymal phenotype with reversal of glutamine production, a leaky cell membrane, and increased ZEB1 to induce EMT (right). ARDS, acute respiratory distress syndrome; Bem, bemcentinib; EMT, epithelial-to-mesenchymal transition; FC, fold change; h, hour; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2.