IPSC screening for drug repurposing identifies anti-RNA virus agents modulating host cell susceptibility.

Keiko Imamura,Jitsutaro Kawaguchi,Yohei Nishi,Takako Enami,Sayaka Okada,Yasuteru Sakurai,Akira Ohta,Ran Shibukawa,Haruhisa Inoue,Thomas Hoenen,Tsugumine Shu,Jiro Yasuda

doi:10.1002/2211-5463.13153

Abstract

Human pathogenic RNA viruses are threats to public health because they are prone to escaping the human immune system through mutations of genomic RNA, thereby causing local outbreaks and global pandemics of emerging or re‐emerging viral diseases. While specific therapeutics and vaccines are being developed, a broad‐spectrum therapeutic agent for RNA viruses would be beneficial for targeting newly emerging and mutated RNA viruses. In this study, we conducted a screen of repurposed drugs using Sendai virus (an RNA virus of the family Paramyxoviridae), with human‐induced pluripotent stem cells (iPSCs) to explore existing drugs that may present anti‐RNA viral activity. Selected hit compounds were evaluated for their efficacy against two important human pathogens: Ebola virus (EBOV) using Huh7 cells and severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) using Vero E6 cells. Selective estrogen receptor modulators (SERMs), including raloxifene, exhibited antiviral activities against EBOV and SARS‐CoV‐2. Pioglitazone, a PPARγ agonist, also exhibited antiviral activities against SARS‐CoV‐2, and both raloxifene and pioglitazone presented a synergistic antiviral effect. Finally, we demonstrated that SERMs blocked entry steps of SARS‐CoV‐2 into host cells. These findings suggest that the identified FDA‐approved drugs can modulate host cell susceptibility against RNA viruses.

Highlights

As previous reports showed that Selective estrogen receptor modulators (SERMs) targeted the host entry step of Ebola virus (EBOV) [21], we investigated whether SERMs inhibited the entry of SARS-CoV-2 into host cells
We conducted compound screening with Sendai virus (SeV) and human induced pluripotent stem cells (iPSCs) for the evaluation of antiviral activity to find a therapeutic candidate against RNA virus-related diseases
We found that several existing drugs including a SERM, anti-tuberculosis, anti-asthmatic, and anti-diabetic drugs suppressed SeV replication

Summary

Ethics statements

The generation and use of human iPSCs were approved by the Ethics Committee of Kyoto University. Human iPSCs, which were generated from a healthy subject (Fig. S1A), were infected with SeV, and an assay system was established to measure viral replication by detecting EGFP-positive iPSCs. We conducted the screening of ~ 500 FDA-approved existing drugs using this system to detect therapeutic agents against. Raloxifene, rifampin, pranlukast, zileuton, and pioglitazone significantly inhibited EGFP mRNA production (Fig. 2A), as shown in remdesivir, RNAdependent RNA polymerase inhibitor approved by FDA for COVID-19 (Fig. S1B) This result indicated that the hit compounds inhibited the RNA synthetic activity of the virus or a step in the viral lifecycle upstream of this process. SERM inhibited the infection of pseudotype VSV with the SARS-CoV spike protein (S) with IC50 values of 4.1, 7.3, 5.8 lM, respectively These results suggested that SERMs harbored antiviral effects by inhibiting the entry of pathogenic coronaviruses into host cells via viral spike proteins (Fig. 5D)

Discussion

Data accessibility