Abstract
Purpose: DNA-dependent protein kinase catalytic subunit (DNA-PKcs, herein referred as DNA-PK) is a multifunctional kinase of high cancer relevance. DNA-PK is deregulated in multiple tumor types, including prostate cancer, and is associated with poor outcomes. DNA-PK was previously nominated as a therapeutic target and DNA-PK inhibitors are currently undergoing clinical investigation. Although DNA-PK is well studied in DNA repair and transcriptional regulation, much remains to be understood about the way by which DNA-PK drives aggressive disease phenotypes. Experimental Design: Here, unbiased proteomic and metabolomic approaches in clinically relevant tumor models uncovered a novel role of DNA-PK in metabolic regulation of cancer progression. DNA-PK regulation of metabolism was interrogated using pharmacologic and genetic perturbation using in vitro cell models, in vivo xenografts, and ex vivo in patient-derived explants (PDE). Results: Key findings reveal: (i) the first-in-field DNA-PK protein interactome; (ii) numerous DNA-PK novel partners involved in glycolysis; (iii) DNA-PK interacts with, phosphorylates (in vitro), and increases the enzymatic activity of glycolytic enzymes ALDOA and PKM2; (iv) DNA-PK drives synthesis of glucose-derived pyruvate and lactate; (v) DNA-PK regulates glycolysis in vitro, in vivo, and ex vivo; and (vi) combination of DNA-PK inhibitor with glycolytic inhibitor 2-deoxyglucose leads to additive anti-proliferative effects in aggressive disease. Conclusions: Findings herein unveil novel DNA-PK partners, substrates, and function in prostate cancer. DNA-PK impacts glycolysis through direct interaction with glycolytic enzymes and modulation of enzymatic activity. These events support energy production that may contribute to generation and/or maintenance of DNA-PK–mediated aggressive disease phenotypes.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 1446-1459 |
| Number of pages | 14 |
| Journal | Clinical Cancer Research |
| Volume | 28 |
| Issue number | 7 |
| DOIs | |
| State | Published - Apr 1 2022 |
Funding
We gratefully thank all the members of the Knudsen laboratory for their intellectual and technical support. We also want to thank the Grabocka Laboratory at Thomas Jefferson University, for their assistance with immunofluorescence experiments and Kayla Bremert for tissue collection through the Australian Prostate Cancer BioResource. Moreover, we want to thank the Translational Pathology core facility at SKCC, and the Bioanalytical Mass Spectrometry Facility (BMSF) at the Mark Wainwright Analytical Centre (MWAC) and UNSW Sydney. In addition, we thank the following institutions that supported this work: the NIH/NCI grants to K.E. Knudsen (5R01CA17640105, 5R01CA18256905) and the Sidney Kimmel Cancer Center (5P30CA056036), the Prostate Cancer Foundation Young Investigator Award to A.A. Shafi, and NCI F99 grant to J.J. McCann (F99CA212225). J. Holst and L.M. Butler are supported by the Tour de Cure Senior Research Grant (RSP-171–18/19). L.M. Butler was supported by a Principal Cancer Research Fellowship produced with the financial and other support of Cancer Council SA’s Beat Cancer Project on behalf of its donors and the State Government of South Australia through the Department of Health. We gratefully thank all the members of the Knudsen laboratory for their intellectual and technical support. We also want to thank the Grabocka Laboratory at Thomas Jefferson University, for their assistance with immunofluorescence experiments and Kayla Bremert for tissue collection through the Australian Prostate Cancer BioResource. Moreover, we want to thank the Translational Pathology core facility at SKCC, and the Bioanalytical Mass Spectrometry Facility (BMSF) at the Mark Wainwright Analytical Centre (MWAC) and UNSW Sydney. In addition, we thank the following institutions that supported this work: the NIH/NCI grants to K.E. Knudsen (5R01CA17640105, 5R01CA18256905) and the Sidney Kimmel Cancer Center (5P30CA056036), the Prostate Cancer Foundation Young Investigator Award to A.A. Shafi, and NCI F99 grant to J.J. McCann (F99CA212225). J. Holst and L.M. Butler are supported by the Tour de Cure Senior Research Grant (RSP-171?18/19). L.M. Butler was supported by a Principal Cancer Research Fellowship produced with the financial and other support of Cancer Council SA?s Beat Cancer Project on behalf of its donors and the State Government of South Australia through the Department of Health.
ASJC Scopus subject areas
- General Medicine
