Mutations in chromatin regulatory proteins have recently been identified in a wide range of human cancers. Although these mutations are associated with disease progression, how they contribute to tumorigenesis remains unknown. We propose to investigate SETD2, a non-redundant histone methyltransferase that preferentially places a trimethylation mark on Histone H3 on lysine 36 in actively transcribed gene bodies. This gene is mutated in approximately 15% of renal cell carcinomas, and is being identified in a growing list of tumors. By examining chromatin organization in primary human tumors, we observed that SETD2 mutation and loss of its histone modification were associated with changes in nucleosome accessibility and widespread alteration in RNA processing. We hypothesize that histone mark dysregulation through SETD2 loss is an important component of tumorigenesis. We propose a highly collaborative project that employs biochemical, genetic and genomic approaches to comprehensively explore the effect of SETD2 mutation on chromatin and transcription, in an effort to unravel the mechanisms by which SETD2 loss leads to the promotion of cancer. To this end, we have generated a unique set of cell-based model systems which uses comparative genomic studies between human and yeast cells. We have additionally established novel domain-specific activities of SETD2, and the first demonstration of functional mutations separating di-methylating from tri-methylating activity. We propose three complementary aims: 1) to define the chromatin reprogramming and transcriptional effects of SETD2 mutation, revealing critical mechanistic features that link histone modification and nucleosome position to the transcriptional and DNA repair defects associated with SETD2 loss, 2) to utilize the separation of function revealed by high severity SETD2 mutants in the catalytic (SET) domain and the RNA polymerase II interactions domain (SRI) to reveal important determinants of the genomic phenotype, and 3) to explore discrete functions associated with SETD2 loss mediated by the altered H3K36me3 mark, by systematically examining the histone reader molecules that transduce signals for DNA methylation, DNA repair, and transcriptional elongation. Taken together, these studies will define the mechanistic link between SETD2 loss of function and cancer development, as well as reveal novel opportunities for biomarker or therapeutic interventions.

Public Health Relevance

This proposal examines the activity of SETD2 mutations in renal cell carcinomas to drive tumor cell biology. Our team has developed abundant preliminary data to support a role for SETD2 (or the yeast ortholog Set2) in maintaining H3K36 trimethylation on gene bodies and suppressing cryptic transcription (yeast) and aberrant transcript processing (human tumors). We propose to examine the structure/function relationship of SETD2 deficiency to the chromatin programming, DNA repair, and transcript processing events mediated by Histone H3 trimethylation to reveal novel mechanisms by which altered chromatin and tumor cell transcript processing promote tumorigenesis.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA198482-06
Application #
9751217
Study Section
Cancer Genetics Study Section (CG)
Program Officer
Okano, Paul
Project Start
2015-09-30
Project End
2020-08-31
Budget Start
2019-09-01
Budget End
2020-08-31
Support Year
6
Fiscal Year
2019
Total Cost
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Type
DUNS #
079917897
City
Nashville
State
TN
Country
United States
Zip Code
37232
Slaughter, Mariesa J; Shanle, Erin K; McFadden, Andrew W et al. (2018) PBRM1 bromodomains variably influence nucleosome interactions and cellular function. J Biol Chem 293:13592-13603
Martin, Donna M; Rathmell, W Kimryn; Tavazoie, Sohail F (2018) Balancing dual demands on the physician-scientist workforce. J Clin Invest 128:3204-3205
Chiang, Yun-Chen; Park, In-Young; Terzo, Esteban A et al. (2018) SETD2 Haploinsufficiency for Microtubule Methylation Is an Early Driver of Genomic Instability in Renal Cell Carcinoma. Cancer Res 78:3135-3146
Panda, Anshuman; de Cubas, Aguirre A; Stein, Mark et al. (2018) Endogenous retrovirus expression is associated with response to immune checkpoint blockade in clear cell renal cell carcinoma. JCI Insight 3:
Kuwahara, Yasumichi; Kennedy, Leslie M; Karnezis, Anthony N et al. (2018) High Frequency of Ovarian Cyst Development in Vhl2B/+;Snf5+/- Mice. Am J Pathol 188:1510-1516
McDaniel, Stephen L; Strahl, Brian D (2017) Shaping the cellular landscape with Set2/SETD2 methylation. Cell Mol Life Sci 74:3317-3334
Murakami, A; Wang, L; Kalhorn, S et al. (2017) Context-dependent role for chromatin remodeling component PBRM1/BAF180 in clear cell renal cell carcinoma. Oncogenesis 6:e287
McDaniel, Stephen L; Hepperla, Austin J; Huang, Jie et al. (2017) H3K36 Methylation Regulates Nutrient Stress Response in Saccharomyces cerevisiae by Enforcing Transcriptional Fidelity. Cell Rep 19:2371-2382
Hacker, Kathryn E; Fahey, Catherine C; Shinsky, Stephen A et al. (2016) Structure/Function Analysis of Recurrent Mutations in SETD2 Protein Reveals a Critical and Conserved Role for a SET Domain Residue in Maintaining Protein Stability and Histone H3 Lys-36 Trimethylation. J Biol Chem 291:21283-21295
Kishton, Rigel J; Barnes, Carson E; Nichols, Amanda G et al. (2016) AMPK Is Essential to Balance Glycolysis and Mitochondrial Metabolism to Control T-ALL Cell Stress and Survival. Cell Metab 23:649-62

Showing the most recent 10 out of 12 publications