l-2-Hydroxyglutarate remodeling of the epigenome and epitranscriptome creates a metabolic vulnerability in kidney cancer models
Anirban Kundu, Garrett J. Brinkley, Hyeyoung Nam, Suman Karki, Richard Kirkman, Madhuparna Pandit, EunHee Shim, Hayley Widden, Juan Liu, Yasaman Heidarian, Nader H. Mahmoudzadeh, Alexander J. Fitt, Devin Absher, Han-Fei Ding, David K. Crossman, William J. Placzek, Jason W. Locasale, Dinesh Rakheja, Jonathan E. McConathy, Rekha Ramachandran, Sejong Bae, Jason M. Tennessen, Sunil Sudarshan
Anirban Kundu, Garrett J. Brinkley, Hyeyoung Nam, Suman Karki, Richard Kirkman, Madhuparna Pandit, EunHee Shim, Hayley Widden, Juan Liu, Yasaman Heidarian, Nader H. Mahmoudzadeh, Alexander J. Fitt, Devin Absher, Han-Fei Ding, David K. Crossman, William J. Placzek, Jason W. Locasale, Dinesh Rakheja, Jonathan E. McConathy, Rekha Ramachandran, Sejong Bae, Jason M. Tennessen, Sunil Sudarshan
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Research Article Metabolism Oncology

l-2-Hydroxyglutarate remodeling of the epigenome and epitranscriptome creates a metabolic vulnerability in kidney cancer models

Abstract

Tumor cells are known to undergo considerable metabolic reprogramming to meet their unique demands and drive tumor growth. At the same time, this reprogramming may come at a cost with resultant metabolic vulnerabilities. The small molecule l-2-hydroxyglutarate (l-2HG) is elevated in the most common histology of renal cancer. Similarly to other oncometabolites, l-2HG has the potential to profoundly impact gene expression. Here, we demonstrate that l-2HG remodels amino acid metabolism in renal cancer cells through combined effects on histone methylation and RNA N6-methyladenosine. The combined effects of l-2HG result in a metabolic liability that renders tumors cells reliant on exogenous serine to support proliferation, redox homeostasis, and tumor growth. In concert with these data, high–l-2HG kidney cancers demonstrate reduced expression of multiple serine biosynthetic enzymes. Collectively, our data indicate that high–l-2HG renal tumors could be specifically targeted by strategies that limit serine availability to tumors.

Authors

Anirban Kundu, Garrett J. Brinkley, Hyeyoung Nam, Suman Karki, Richard Kirkman, Madhuparna Pandit, EunHee Shim, Hayley Widden, Juan Liu, Yasaman Heidarian, Nader H. Mahmoudzadeh, Alexander J. Fitt, Devin Absher, Han-Fei Ding, David K. Crossman, William J. Placzek, Jason W. Locasale, Dinesh Rakheja, Jonathan E. McConathy, Rekha Ramachandran, Sejong Bae, Jason M. Tennessen, Sunil Sudarshan

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Figure 2

l-2HG suppression of ATF4.

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l-2HG suppression of ATF4. (A) Immunoblot for ATF4 protein from 786-O, ...
(A) Immunoblot for ATF4 protein from 786-O, OS-RC-2, and 769p RCC cells stably transduced with the indicated vectors. Actin was used as loading control. (B) Immunoblot for ATF4 protein from 786-O cells stably transduced with control vector, L2HGDH (WT), or L2HGDH A241G (mutant). Actin was used as loading control. (C) Immunoblot for L2HGDH and ATF4 protein from control and L2HGDH-KO HK-2 cells. Actin was used as loading control. (D) Immunoblot for ATF4 and PHGDH from 769p cells stably expressing either control vector or ATF4 cDNA. Actin was used as loading control. (E) Relative ATF4 mRNA normalized to TBP was examined by RT-qPCR from 769p and 786-O cells stably expressing either control (black) vector or L2HGDH cDNA (red). Data are expressed as mean ± SD from n = 3 (769p) or n = 4 (786-O) biological replicates. *P < 0.05, **P < 0.005. (F and G) 786-O (F) and A498 (G) cells stably transduced with L2HGDH cDNA were transiently transfected (55 hours) with either scramble siRNA (Scr) or siRNAs targeting KDM4C (#1, #2). Immunoblotting for KDM4C, ATF4, PHGDH, and PSAT1 was performed. Actin (β-actin) or tubulin (α-tubulin) was used as loading control. (H) Top: The construct containing the human ATF4 5′-uORFs preceding firefly luciferase. Bottom: Relative luciferase signal following transient transfection of the luciferase construct into 769p and 786-O cells stably expressing the indicated vector (control vector, black; L2HGDH WT, red; or L2HGDH A141G mutant, blue). Data were normalized to Renilla luciferase (Luc). Data are presented as mean ± SEM. ANOVA was used, and Tukey’s post hoc P values are shown.