Cargando…

Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer

To identify novel drivers of malignancy in pancreatic ductal adenocarcinoma (PDAC), we employed regulatory network analysis, which calculates the activity of transcription factors and other regulatory proteins based on the integrated expression of their positive and negative target genes. We generat...

Descripción completa

Detalles Bibliográficos
Autores principales: Maurer, H. Carlo, Curiel-Garcia, Alvaro, Holmstrom, Sam, Laise, Pasquale, Palermo, Carmine F., Sastra, Steven A., Andren, Anthony, Li, Zhang, LeLarge, Tessa, Sagalovskiy, Irina, Ross, Daniel R., Rosario, Vilma, Lu, Kate, Ferraiuolo, Ethan, Spinosa, Nicholas, Wong, Winston, Shaw, Kaitlin, Chabot, John A., Genkinger, Jeanine, Hibshoosh, Hanina, Manji, Gulam A., Iuga, Alina, Schmid, Roland M., Badgley, Michael A., Johnson, Kristen, Califano, Andrea, Lyssiotis, Costas, Olive, Kenneth P.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10055246/
https://www.ncbi.nlm.nih.gov/pubmed/36993718
http://dx.doi.org/10.1101/2023.03.19.533333
_version_ 1785015844541038592
author Maurer, H. Carlo
Curiel-Garcia, Alvaro
Holmstrom, Sam
Laise, Pasquale
Palermo, Carmine F.
Sastra, Steven A.
Andren, Anthony
Li, Zhang
LeLarge, Tessa
Sagalovskiy, Irina
Ross, Daniel R.
Rosario, Vilma
Lu, Kate
Ferraiuolo, Ethan
Spinosa, Nicholas
Wong, Winston
Shaw, Kaitlin
Chabot, John A.
Genkinger, Jeanine
Hibshoosh, Hanina
Manji, Gulam A.
Iuga, Alina
Schmid, Roland M.
Badgley, Michael A.
Johnson, Kristen
Califano, Andrea
Lyssiotis, Costas
Olive, Kenneth P.
author_facet Maurer, H. Carlo
Curiel-Garcia, Alvaro
Holmstrom, Sam
Laise, Pasquale
Palermo, Carmine F.
Sastra, Steven A.
Andren, Anthony
Li, Zhang
LeLarge, Tessa
Sagalovskiy, Irina
Ross, Daniel R.
Rosario, Vilma
Lu, Kate
Ferraiuolo, Ethan
Spinosa, Nicholas
Wong, Winston
Shaw, Kaitlin
Chabot, John A.
Genkinger, Jeanine
Hibshoosh, Hanina
Manji, Gulam A.
Iuga, Alina
Schmid, Roland M.
Badgley, Michael A.
Johnson, Kristen
Califano, Andrea
Lyssiotis, Costas
Olive, Kenneth P.
author_sort Maurer, H. Carlo
collection PubMed
description To identify novel drivers of malignancy in pancreatic ductal adenocarcinoma (PDAC), we employed regulatory network analysis, which calculates the activity of transcription factors and other regulatory proteins based on the integrated expression of their positive and negative target genes. We generated a regulatory network for the malignant epithelial cells of human PDAC using gene expression data from a set of 197 laser capture microdissected human PDAC samples and 45 low-grade precursors, for which we had matched histopathological, clinical, and epidemiological annotation. We then identified the most highly activated and repressed regulatory proteins (e.g. master regulators or MRs) associated with four malignancy phenotypes: precursors vs. PDAC (initiation), low-grade vs. high grade histopathology (progression), survival post resection, and association with KRAS activity. Integrating across these phenotypes, the top MR of PDAC malignancy was found to be BMAL2, a member of the PAS family of bHLH transcription factors. Although the canonical function of BMAL2 is linked to the circadian rhythm protein CLOCK, annotation of BMAL2 target genes highlighted a potential role in hypoxia response. We previously demonstrated that PDAC is hypovascularized and hypoperfused, and here show that PDAC from the genetically engineered KPC model exists in a state of extreme hypoxia, with a partial oxygen pressure of <1mmHg. Given the close homology of BMAL2 to HIF1β (ARNT) and its potential to heterodimerize with HIF1A and HIF2A, we investigated whether BMAL2 plays a role in the hypoxic response of PDAC. Indeed, BMAL2 controlled numerous hypoxia response genes and could be inhibited following treatment with multiple RAF, MEK, and ERK inhibitors, validating its association with RAS activity. Knockout of BMAL2 in four human PDAC cell lines led to defects in growth and invasion in the setting of hypoxia. Strikingly, BMAL2 null cells failed to induce glycolysis upon exposure to severe hypoxia and this was associated with a loss of expression of the glycolytic enzyme LDHA. Moreover, HIF1A was no longer stabilized under hypoxia in BMAL2 knockout cells. By contrast, HIF2A was hyper-stabilized under hypoxia, indicating a dysregulation of hypoxia metabolism in response to BMAL2 loss. We conclude that BMAL2 is a master regulator of hypoxic metabolism in PDAC, serving as a molecular switch between the disparate metabolic roles of HIF1A- and HIF2A-dependent hypoxia responses.
format Online
Article
Text
id pubmed-10055246
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Cold Spring Harbor Laboratory
record_format MEDLINE/PubMed
spelling pubmed-100552462023-03-30 Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer Maurer, H. Carlo Curiel-Garcia, Alvaro Holmstrom, Sam Laise, Pasquale Palermo, Carmine F. Sastra, Steven A. Andren, Anthony Li, Zhang LeLarge, Tessa Sagalovskiy, Irina Ross, Daniel R. Rosario, Vilma Lu, Kate Ferraiuolo, Ethan Spinosa, Nicholas Wong, Winston Shaw, Kaitlin Chabot, John A. Genkinger, Jeanine Hibshoosh, Hanina Manji, Gulam A. Iuga, Alina Schmid, Roland M. Badgley, Michael A. Johnson, Kristen Califano, Andrea Lyssiotis, Costas Olive, Kenneth P. bioRxiv Article To identify novel drivers of malignancy in pancreatic ductal adenocarcinoma (PDAC), we employed regulatory network analysis, which calculates the activity of transcription factors and other regulatory proteins based on the integrated expression of their positive and negative target genes. We generated a regulatory network for the malignant epithelial cells of human PDAC using gene expression data from a set of 197 laser capture microdissected human PDAC samples and 45 low-grade precursors, for which we had matched histopathological, clinical, and epidemiological annotation. We then identified the most highly activated and repressed regulatory proteins (e.g. master regulators or MRs) associated with four malignancy phenotypes: precursors vs. PDAC (initiation), low-grade vs. high grade histopathology (progression), survival post resection, and association with KRAS activity. Integrating across these phenotypes, the top MR of PDAC malignancy was found to be BMAL2, a member of the PAS family of bHLH transcription factors. Although the canonical function of BMAL2 is linked to the circadian rhythm protein CLOCK, annotation of BMAL2 target genes highlighted a potential role in hypoxia response. We previously demonstrated that PDAC is hypovascularized and hypoperfused, and here show that PDAC from the genetically engineered KPC model exists in a state of extreme hypoxia, with a partial oxygen pressure of <1mmHg. Given the close homology of BMAL2 to HIF1β (ARNT) and its potential to heterodimerize with HIF1A and HIF2A, we investigated whether BMAL2 plays a role in the hypoxic response of PDAC. Indeed, BMAL2 controlled numerous hypoxia response genes and could be inhibited following treatment with multiple RAF, MEK, and ERK inhibitors, validating its association with RAS activity. Knockout of BMAL2 in four human PDAC cell lines led to defects in growth and invasion in the setting of hypoxia. Strikingly, BMAL2 null cells failed to induce glycolysis upon exposure to severe hypoxia and this was associated with a loss of expression of the glycolytic enzyme LDHA. Moreover, HIF1A was no longer stabilized under hypoxia in BMAL2 knockout cells. By contrast, HIF2A was hyper-stabilized under hypoxia, indicating a dysregulation of hypoxia metabolism in response to BMAL2 loss. We conclude that BMAL2 is a master regulator of hypoxic metabolism in PDAC, serving as a molecular switch between the disparate metabolic roles of HIF1A- and HIF2A-dependent hypoxia responses. Cold Spring Harbor Laboratory 2023-03-21 /pmc/articles/PMC10055246/ /pubmed/36993718 http://dx.doi.org/10.1101/2023.03.19.533333 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Maurer, H. Carlo
Curiel-Garcia, Alvaro
Holmstrom, Sam
Laise, Pasquale
Palermo, Carmine F.
Sastra, Steven A.
Andren, Anthony
Li, Zhang
LeLarge, Tessa
Sagalovskiy, Irina
Ross, Daniel R.
Rosario, Vilma
Lu, Kate
Ferraiuolo, Ethan
Spinosa, Nicholas
Wong, Winston
Shaw, Kaitlin
Chabot, John A.
Genkinger, Jeanine
Hibshoosh, Hanina
Manji, Gulam A.
Iuga, Alina
Schmid, Roland M.
Badgley, Michael A.
Johnson, Kristen
Califano, Andrea
Lyssiotis, Costas
Olive, Kenneth P.
Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer
title Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer
title_full Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer
title_fullStr Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer
title_full_unstemmed Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer
title_short Ras-dependent activation of BMAL2 regulates hypoxic metabolism in pancreatic cancer
title_sort ras-dependent activation of bmal2 regulates hypoxic metabolism in pancreatic cancer
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10055246/
https://www.ncbi.nlm.nih.gov/pubmed/36993718
http://dx.doi.org/10.1101/2023.03.19.533333
work_keys_str_mv AT maurerhcarlo rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT curielgarciaalvaro rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT holmstromsam rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT laisepasquale rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT palermocarminef rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT sastrastevena rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT andrenanthony rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT lizhang rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT lelargetessa rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT sagalovskiyirina rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT rossdanielr rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT rosariovilma rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT lukate rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT ferraiuoloethan rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT spinosanicholas rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT wongwinston rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT shawkaitlin rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT chabotjohna rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT genkingerjeanine rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT hibshooshhanina rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT manjigulama rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT iugaalina rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT schmidrolandm rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT badgleymichaela rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT johnsonkristen rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT califanoandrea rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT lyssiotiscostas rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer
AT olivekennethp rasdependentactivationofbmal2regulateshypoxicmetabolisminpancreaticcancer