TY - JOUR
T1 - HIF2 Regulates Intestinal Wnt5a Expression
AU - García García, Carolina J.
AU - Acevedo Diaz, Ariana C.
AU - Kumari, Neeraj
AU - Govindaraju, Suman
AU - de la Cruz Bonilla, Marimar
AU - San Lucas, F. Anthony
AU - Nguyen, Nicholas D.
AU - Jiménez Sacarello, Iancarlos
AU - Piwnica-Worms, Helen
AU - Maitra, Anirban
AU - Taniguchi, Cullen M.
N1 - Funding Information:
CT was supported by funding from the National Institutes of Health (NIH) under award number R01CA227517-01A1 and GI SPORE grant (P50CA221707), by the Cancer Prevention & Research Institute of Texas (CPRIT) under grant RR140012, by the V Foundation (V2015-22), by the Sidney Kimmel Foundation, by a Sabin Family Foundation Fellowship, by the Reaumond Family Foundation, by the Mark Foundation, by the Childress Family Foundation, by the McNair Family Foundation, and by generous philanthropic contributions to The University of Texas MD Anderson Moon Shots Program. CG was supported by the National Institute of Diabetes, Digestive and Kidney Diseases (NIDDK) of the NIH under award number F31DK121384. MD was supported by the National Cancer Institute (NCI) of the NIH under award number F31CA210631. CG, MD, AA, and IS were also supported by the NIH/NCI under award number U54CA096300/297. This work was also supported in part by NIH/NIDDK grant DK056338, which supports the Texas Medical Center Digestive Diseases Center, and by NIH/NCI Cancer Center Support Grants (CCSG) P30CA016672, which supports MDACC’s Sequencing and Microarray Facility.
Publisher Copyright:
Copyright © 2021 García García, Acevedo Diaz, Kumari, Govindaraju, de la Cruz Bonilla, San Lucas, Nguyen, Jiménez Sacarello, Piwnica-Worms, Maitra and Taniguchi.
PY - 2021/11/25
Y1 - 2021/11/25
N2 - Radiation therapy for abdominal tumors is challenging because the small intestine is exquisitely radiosensitive. Unfortunately, there are no FDA-approved therapies to prevent or mitigate GI radiotoxicity. The EGLN protein family are oxygen sensors that regulate cell survival and metabolism through the degradation of hypoxia-inducible factors (HIFs). Our group has previously shown that stabilization of HIF2 through genetic deletion or pharmacologic inhibition of the EGLNs mitigates and protects against GI radiotoxicity in mice by improving intestinal crypt stem cell survival. Here we aimed to elucidate the molecular mechanisms by which HIF2 confers GI radioprotection. We developed duodenal organoids from mice, transiently overexpressed non-degradable HIF2, and performed bulk RNA sequencing. Interestingly, HIF2 upregulated known radiation modulators and genes involved in GI homeostasis, including Wnt5a. Non-canonical Wnt5a signaling has been shown by other groups to improve intestinal crypt regeneration in response to injury. Here we show that HIF2 drives Wnt5a expression in multiple duodenal organoid models. Luciferase reporter assays performed in human cells showed that HIF2 directly activates the WNT5A promoter via a hypoxia response element. We then evaluated crypt regeneration using spheroid formation assays. Duodenal organoids that were pre-treated with recombinant Wnt5a had a higher cryptogenic capacity after irradiation, compared to vehicle-treated organoids. Conversely, we found that Wnt5a knockout decreased the cryptogenic potential of intestinal stem cells following irradiation. Treatment with recombinant Wnt5a prior to irradiation rescued the cryptogenic capacity of Wnt5a knockout organoids, indicating that Wnt5a is necessary and sufficient for duodenal radioprotection. Taken together, our results suggest that HIF2 radioprotects the GI tract by inducing Wnt5a expression.
AB - Radiation therapy for abdominal tumors is challenging because the small intestine is exquisitely radiosensitive. Unfortunately, there are no FDA-approved therapies to prevent or mitigate GI radiotoxicity. The EGLN protein family are oxygen sensors that regulate cell survival and metabolism through the degradation of hypoxia-inducible factors (HIFs). Our group has previously shown that stabilization of HIF2 through genetic deletion or pharmacologic inhibition of the EGLNs mitigates and protects against GI radiotoxicity in mice by improving intestinal crypt stem cell survival. Here we aimed to elucidate the molecular mechanisms by which HIF2 confers GI radioprotection. We developed duodenal organoids from mice, transiently overexpressed non-degradable HIF2, and performed bulk RNA sequencing. Interestingly, HIF2 upregulated known radiation modulators and genes involved in GI homeostasis, including Wnt5a. Non-canonical Wnt5a signaling has been shown by other groups to improve intestinal crypt regeneration in response to injury. Here we show that HIF2 drives Wnt5a expression in multiple duodenal organoid models. Luciferase reporter assays performed in human cells showed that HIF2 directly activates the WNT5A promoter via a hypoxia response element. We then evaluated crypt regeneration using spheroid formation assays. Duodenal organoids that were pre-treated with recombinant Wnt5a had a higher cryptogenic capacity after irradiation, compared to vehicle-treated organoids. Conversely, we found that Wnt5a knockout decreased the cryptogenic potential of intestinal stem cells following irradiation. Treatment with recombinant Wnt5a prior to irradiation rescued the cryptogenic capacity of Wnt5a knockout organoids, indicating that Wnt5a is necessary and sufficient for duodenal radioprotection. Taken together, our results suggest that HIF2 radioprotects the GI tract by inducing Wnt5a expression.
KW - GI radiotoxicity
KW - HIF2
KW - Wnt5a
KW - hypoxia
KW - intestinal stem cells
KW - radiotherapy
UR - http://www.scopus.com/inward/record.url?scp=85120905493&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85120905493&partnerID=8YFLogxK
U2 - 10.3389/fonc.2021.769385
DO - 10.3389/fonc.2021.769385
M3 - Article
C2 - 34900719
AN - SCOPUS:85120905493
SN - 2234-943X
VL - 11
JO - Frontiers in Oncology
JF - Frontiers in Oncology
M1 - 769385
ER -