Modifications of lipid pathways restrict SARS-CoV-2 propagation in human induced pluripotent stem cell-derived 3D airway organoids

Ping-Hsing Tsai; Jun-Ren Sun; Yueh Chien; Man Sheung Chan; Winnie Khor; Hsin-Chou Yang; Chih-Heng Huang; Chia-Ni Hsiung; Teh-Yang Hwa; Yi-Ying Lin; Chih-Ling Yeh; Mong-Lien Wang; Yi-Ping Yang; Yuh-Min Chen; Fu-Ting Tsai; Meng-Shiue Lee; Yun-Hsiang Cheng; Shan-Ko Tsai; Ping-Cheng Liu; Shih-Jie Chou; Shih-Hwa Chiou

doi:10.1016/j.jare.2023.08.005

Modifications of lipid pathways restrict SARS-CoV-2 propagation in human induced pluripotent stem cell-derived 3D airway organoids

J Adv Res. 2023 Aug 7:S2090-1232(23)00209-6. doi: 10.1016/j.jare.2023.08.005. Online ahead of print.

Authors

Ping-Hsing Tsai¹, Jun-Ren Sun², Yueh Chien³, Man Sheung Chan³, Winnie Khor³, Hsin-Chou Yang⁴, Chih-Heng Huang⁵, Chia-Ni Hsiung⁴, Teh-Yang Hwa⁴, Yi-Ying Lin³, Chih-Ling Yeh³, Mong-Lien Wang⁶, Yi-Ping Yang⁶, Yuh-Min Chen⁷, Fu-Ting Tsai³, Meng-Shiue Lee³, Yun-Hsiang Cheng², Shan-Ko Tsai⁸, Ping-Cheng Liu⁹, Shih-Jie Chou¹⁰, Shih-Hwa Chiou¹¹

Affiliations

¹ Department of Medical Research, Taipei Veterans General Hospital, Taipei 11217, Taiwan; Institute of Pharmacology, School of Medicine, National Yang Ming Chiao Tung University, Taipei 112304, Taiwan.
² Institute of Preventive Medicine, National Defense Medical Center, Taipei 11217, Taiwan; Graduate Institute of Medical Science, National Defense Medical Center, Taipei, Taiwan; Department of Physiology and Biophysics, Graduate Institute of Physiology, National Defense Medical Center, Taipei, Taiwan.
³ Department of Medical Research, Taipei Veterans General Hospital, Taipei 11217, Taiwan.
⁴ Institute of Statistical Science, Academia Sinica, Taipei 11529, Taiwan.
⁵ Institute of Preventive Medicine, National Defense Medical Center, Taipei 11217, Taiwan; Department of Microbiology and Immunology, National Defense Medical Center, Taipei 11217, Taiwan; Graduate Institute of Medical Science, National Defense Medical Center, Taipei, Taiwan.
⁶ Department of Medical Research, Taipei Veterans General Hospital, Taipei 11217, Taiwan; Institute of Food Safety and Health Risk Assessment, School of Pharmaceutical Sciences, National Yang Ming Chiao Tung University, Taipei 112304, Taiwan.
⁷ Department of Chest Medicine, Taipei Veterans General Hospital, Taipei 112, Taiwan; Faculty of Medicine, School of Medicine, National Yang Ming Chiao Tung University, Taipei 112, Taiwan.
⁸ Institute of Preventive Medicine, National Defense Medical Center, Taipei 11217, Taiwan.
⁹ Department of Microbiology and Immunology, National Defense Medical Center, Taipei 11217, Taiwan; Graduate Institute of Applied Science and Technology, National Taiwan University of Science and Technology, Taipei, Taiwan.
¹⁰ Department of Medical Research, Taipei Veterans General Hospital, Taipei 11217, Taiwan; Institute of Pharmacology, School of Medicine, National Yang Ming Chiao Tung University, Taipei 112304, Taiwan. Electronic address: sjchou3@vghtpe.gov.tw.
¹¹ Department of Medical Research, Taipei Veterans General Hospital, Taipei 11217, Taiwan; Institute of Pharmacology, School of Medicine, National Yang Ming Chiao Tung University, Taipei 112304, Taiwan. Electronic address: shchiou@vghtpe.gov.tw.

PMID: 37557954
DOI: 10.1016/j.jare.2023.08.005

Abstract

Background: Modifications of lipid metabolism were closely associated with the manifestations and prognosis of coronavirus disease of 2019 (COVID-19). Pre-existing metabolic conditions exacerbated the severity of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection while modulations of aberrant lipid metabolisms alleviated the manifestations. To elucidate the underlying mechanisms, an experimental platform that reproduces human respiratory physiology is required.

Methods: Here we generated induced pluripotent stem cell-derived airway organoids (iPSC-AOs) that resemble the human native airway. Single-cell sequencing (ScRNAseq) and microscopic examination verified the cellular heterogeneity and microstructures of iPSC-AOs, respectively. We subjected iPSC-AOs to SARS-CoV-2 infection and investigated the treatment effect of lipid modifiers statin drugs on viral pathogenesis, gene expression, and the intracellular trafficking of the SARS-CoV-2 entry receptor angiotensin-converting enzyme-2 (ACE-2).

Results: In SARS-CoV-2-infected iPSC-AOs, immunofluorescence staining detected the SARS-CoV-2 spike (S) and nucleocapsid (N) proteins and bioinformatics analysis further showed the aberrant enrichment of lipid-associated pathways. In addition, SARS-CoV-2 hijacked the host RNA replication machinery and generated the new isoforms of a high-density lipoprotein constituent apolipoprotein A1 (APOA1) and the virus-scavenging protein deleted in malignant brain tumors 1 (DMBT1). Manipulating lipid homeostasis using cholesterol-lowering drugs (e.g. Statins) relocated the viral entry receptor angiotensin-converting enzyme-2 (ACE-2) and decreased N protein expression, leading to the reduction of SARS-CoV-2 entry and replication. The same lipid modifications suppressed the entry of luciferase-expressing SARS-CoV-2 pseudoviruses containing the S proteins derived from different SARS-CoV-2 variants, i.e. wild-type, alpha, delta, and omicron.

Conclusions: Together, our data demonstrated that modifications of lipid pathways restrict SARS-CoV-2 propagation in the iPSC-AOs, which the inhibition is speculated through the translocation of ACE2 from the cell membrane to the cytosol. Considering the highly frequent mutation and generation of SARS-CoV-2 variants, targeting host metabolisms of cholesterol or other lipids may represent an alternative approach against SARS-CoV-2 infection.

Keywords: Airway organoid; Angiotensin-converting enzyme 2; Induced pluripotent stem cell; Severe acute respiratory syndrome coronavirus 2; Single cell RNA-sequencing.