key: cord-0736037-q08y3lxh authors: Deguchi, Sayaka; Serrano‐Aroca, Ángel; Tambuwala, Murtaza M.; Uhal, Bruce D.; Brufsky, Adam M.; Takayama, Kazuo title: SARS‐CoV‐2 research using human pluripotent stem cells and organoids date: 2021-07-24 journal: Stem Cells Transl Med DOI: 10.1002/sctm.21-0183 sha: b2af501ddf24fed44a34e2739722432ff719d572 doc_id: 736037 cord_uid: q08y3lxh Experimental cell models are indispensable for clarifying the pathophysiology of coronavirus disease 2019 (COVID‐19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection, and for developing therapeutic agents. To recapitulate the symptoms and drug response of COVID‐19 patients in vitro, SARS‐CoV‐2 studies using physiologically relevant human embryonic stem (ES)/induced pluripotent stem (iPS) cell‐derived somatic cells and organoids are ongoing. These cells and organoids have been used to show that SARS‐CoV‐2 can infect and damage various organs including the lung, heart, brain, intestinal tract, kidney, and pancreas. They are also being used to develop COVID‐19 therapeutic agents, including evaluation of their antiviral efficacy and safety. The relationship between COVID‐19 aggravation and human genetic backgrounds has been investigated using genetically modified ES/iPS cells and patient‐derived iPS cells. This review summarizes the latest results and issues of SARS‐CoV‐2 research using human ES/iPS cell‐derived somatic cells and organoids. ischemia, diarrhea, stroke, acute kidney injury, and hyperglycemia. Elucidation of the complex pathophysiology of COVID-19 and the development of therapeutic agents are essential for stopping this pandemic. While several COVID-19 vaccines are now available, progress in therapeutic agents has been slower. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the cause of COVID-19, uses its spike (S) protein to enter host cells. Because the viral S protein binds to angiotensin converting enzyme 2 (ACE2) on the host cell surface, cells expressing ACE2 are susceptible to SARS-CoV-2 infection. Accordingly, SARS-CoV-2 has high infectivity in primates, including humans, rhesus monkeys, and cynomolgus monkeys, but low infectivity in wild-type mice, [1] [2] [3] [4] limiting the animal species that can be used for experiments. Moreover, it is ethically difficult to use these animals in large quantities. Therefore, physiologically relevant human embryonic stem (ES)/induced pluripotent stem (iPS) cell-derived somatic cells and organoids are being developed as cell models for SARS-CoV-2 infection. Organoids are three-dimensional structures that can be generated from somatic stem cells 5 or human ES/iPS cells. 6 The derived somatic cells and organoids have closer cellular and organ functions to primary cells than other cell lines commonly used in in vitro SARS-CoV-2 research, such as Vero, Calu-3, and Caco-2, suggesting that they more accurately reproduce the pathophysiology of COVID-19 and drug effects. 7 Although the infection efficiency of SARS-CoV-2 in Vero, Calu-3, and Caco-2 cells, it is difficult to reproduce the cellular and organ responses due to SARS-CoV-2 infection. This review introduces the latest findings and issues of SARS-CoV-2 research using human ES/iPS cell-derived somatic cells and organoids. The main symptoms of COVID-19 are manifested in the respiratory system, but many cases of multiple organ failure have been reported, [8] [9] [10] indicating many organs are affected by SARS-CoV-2 infection. Accordingly, human ES/iPS cell-derived somatic cells and organoids have been used to study the infection in several of these organs (Table 1) . Because respiratory failure is one of the most critical symptoms of SARS-CoV-2 infection, experiments using bronchial and alveolar models are especially being studied. We generated human bronchial organoids from cryopreserved human bronchial epithelial cells to reproduce the infection of SARS-CoV-2 in the bronchi. 22 These bronchial organoids have cellular constituents resembling basal, ciliated, goblet, and club cells, and we confirmed that some basal cells can be infected with SARS-CoV-2. Pei et al also performed SARS-CoV-2 infection experiments using airway organoids generated from human ES cells, finding ciliated and club cells are susceptible to infection. 14 Remdesivir also reduced the intracellular viral genome five magnitudes. 24 Remdesivir was shown to have an antiviral effect on human ES cell-derived intestinal organoids, with the intracellular viral genome level reduced four magnitudes. 29 To inhibit the infection of SARS-CoV-2, a soluble recombinant protein of human ACE2, which is a SARS-CoV-2 receptor, has been used. Monteil et al showed that treating human iPS cell-derived capillary organoids and human ES cell-derived kidney organoids with human recombinant soluble ACE2 (hrsACE2) can inhibit SARS-CoV-2 infection. 32 Bojkova et al also reported that intracellular viral genome levels in human iPS cell-derived cardiomyocytes are reduced by recombinant SARS-CoV-2 RESEARCH USING PSCs AND ORGANOIDS ACE2 treatment. 23 Following these findings, Aperion Biologics is conducting a phase 2 trial on human recombinant soluble ACE2. Camostat and Nafamostat can inhibit type II transmembrane serine protease (TMPRSS2) to prevent SARS-CoV-2 entry. 41 We reported that Camostat treatment reduced the number of viral genome copies found in cell supernatants derived from cryopreserved human bronchial epithelial cell-derived bronchial organoids to approximately one-twentieth that in the untreated group. 22 In addition, they are expected to assist in searches for antiinflammatory drugs. Furthermore, they can be used to evaluate the toxicity and safety of COVID-19 therapeutic agents. About 80% of COVID-19 patients are asymptomatic or mild, but the other 20% become severe. Various factors, such as aging, medical history, racial differences, and genetics, are predicted in COVID-19 aggravation. [42] [43] [44] [45] Genome-wide association studies performed on mild and severe COVID-19 patients found differences in genetic backgrounds. 46 Because human iPS cells can be established from individuals of any genetic background, they make an attractive model to study the relationship between genetic factors and COVID-19 severity. 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