Summary of Study ST001921

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR001212. The data can be accessed directly via it's Project DOI: 10.21228/M81X3N This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

Study ID	ST001921
Study Title	An Airway Organoid-Based Screen Identifies a Role for the HIF1α-Glycolysis Axis in SARS-CoV-2 Infection
Study Summary	SARS-CoV-2 causes the COVID-19 pandemic. It is urgent to develop disease models to dissect mechanisms regulating SARS-CoV-2 infection. Here, we derive airway organoids from human pluripotent stem cells (hPSC-AOs). The hPSC-AOs, particularly ciliated-like cells, are permissive to SARS-CoV-2 infection. Using this platform, we perform a high content screen and identify GW6471, which blocks SARS-CoV-2 infection. GW6471 can also block infection of the B.1.351 SARS-CoV-2 variant. RNA-seq analysis suggests that GW6471 blocks SARS-CoV-2 infection at least in part by inhibiting HIF1α, which is further validated by chemical inhibitor and genetic perturbation targeting HIF1α. Metabolic profiling identifies decreased rates of glycolysis upon GW6471 treatment, consistent with transcriptome profiling. Finally, xanthohumol, 5-(Tetradecyloxy)-2-furoic acid, and ND-646, three compounds that suppress fatty acid biosynthesis, also block SARS-CoV-2 infection. Together, a high content screen coupled with transcriptome and metabolic profiling reveals a key role of the HIF1α-glycolysis axis in mediating SARS-CoV-2 infection of human airway epithelium.
Institute	Weill Cornell Medicine
Last Name	Chen
First Name	Shuibing
Address	A 827B, 1300 York Ave
Email	shc2034@med.cornell.edu
Phone	2127465431
Submit Date	2021-09-24
Raw Data Available	Yes
Raw Data File Type(s)	raw(Waters)
Analysis Type Detail	GC-MS
Release Date	2021-10-20
Release Version	1

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Treatment:

Treatment ID:	TR002011
Treatment Summary:	SARS-CoV-2 variants, including USA-WA1/2020 and B.1.351, were obtained from the World Reference Center for Emerging Viruses and Arboviruses located at the University of Texas, Medical Branch via the CDC. SARS-CoV-2 was propagated in Vero E6 cells (ATCC) in EMEM supplemented with 10% FCS, 1 mM Sodium Pyruvate and 10 mM HEPES. hPSC-AOs were fragmented into small cell clusters and plated on 10% matrigel-coated plates. The infection was performed in the culture media at the indicated MOIs at 37°C. For pre-infection treatment experiments, hPSC-AOs were pretreated with DMSO (control), 10 µM GW6471 for 4 hours prior to infection. HIF1a knockdown in hPSC-AOs Two shRNAs against HIF1α and one scrambled negative control non-effective shRNA in lentiviral GFP vectors were purchased from OriGene company. The shRNA sequences are shown in Table S3. To generate lentivirus particles, shRNA vectors and lentivirus packaging vectors were co-transfected into 293T cells. The day 15 lung progenitor cells were infected with the collected lentivirus (MOI=0.5) with 8 μg/ml polybrene. To increase the infection efficiency, the cells were centrifuged at 2300 rpm for 1 hour at 30℃. At 24 hpi, the cells were selected with 1 μg/ml puromycin for an additional 72 hours. After selection, the cells were dissociated into single cells and seeded into 24 well plates at 300-400 cells/μl for 3D organoid differentiation.

Treatment ID:

TR002011

Treatment Summary:

SARS-CoV-2 variants, including USA-WA1/2020 and B.1.351, were obtained from the World Reference Center for Emerging Viruses and Arboviruses located at the University of Texas, Medical Branch via the CDC. SARS-CoV-2 was propagated in Vero E6 cells (ATCC) in EMEM supplemented with 10% FCS, 1 mM Sodium Pyruvate and 10 mM HEPES. hPSC-AOs were fragmented into small cell clusters and plated on 10% matrigel-coated plates. The infection was performed in the culture media at the indicated MOIs at 37°C. For pre-infection treatment experiments, hPSC-AOs were pretreated with DMSO (control), 10 µM GW6471 for 4 hours prior to infection. HIF1a knockdown in hPSC-AOs Two shRNAs against HIF1α and one scrambled negative control non-effective shRNA in lentiviral GFP vectors were purchased from OriGene company. The shRNA sequences are shown in Table S3. To generate lentivirus particles, shRNA vectors and lentivirus packaging vectors were co-transfected into 293T cells. The day 15 lung progenitor cells were infected with the collected lentivirus (MOI=0.5) with 8 μg/ml polybrene. To increase the infection efficiency, the cells were centrifuged at 2300 rpm for 1 hour at 30℃. At 24 hpi, the cells were selected with 1 μg/ml puromycin for an additional 72 hours. After selection, the cells were dissociated into single cells and seeded into 24 well plates at 300-400 cells/μl for 3D organoid differentiation.