The p53 transcription factor plays a pivotal role in tumorigenesis. p53 inactivation in tumors, which occurs typically through missense mutation, unequivocally promotes cancer, but the mutated p53 protein also displays gain-of-function (GOF) properties that promote cancer. In this proposal, we strive to deconstruct the transcriptional programs through which wild-type p53 suppresses cancer and through which missense mutant p53 exerts GOF effects to promote cancer. We propose to use integrated genetic, genomic, cell biological and biochemical approaches to define the p53 transcriptional programs critical for p53-mediated suppression of pancreatic cancer, a deadly cancer with a mere 6% 5-year survival rate and which is typically associated with p53 mutation. Our proposed work builds on initial studies of a unique panel of p53 transcriptional activation domain (TAD) mutant knock-in mice that we generated to define the downstream p53 transcriptional targets most essential for suppressing cancer development. A particularly powerful mutant is the TAD1 mutant, known as p5325,26, which is severely compromised for activation of most known p53 target genes yet still efficiently transactivates a small set of novel p53-dependent genes in fibroblasts and still retains full activity in suppressing a variety of cancers. In addition, the TD2 mutant, p5353,54 can hyperactivate a subset of p53 target genes and behaves as a super-tumor suppressor. The p53 tumor suppression associated target genes (TSAGs) activated by the p5325,26 and p5353,54 mutants largely represent novel p53 targets, which we hypothesize are critical for tumor suppression and therefore have great potential to expand our current knowledge of p53 tumor suppression mechanisms. Here, we propose to perform ChIP-seq and RNA-seq in premalignant pancreatic ductal epithelial cells expressing these mutants to identify TSAGs associated with pancreatic cancer suppression. We then propose to identify key mediators of p53 function in tumor suppression by performing CRISPR screening in mouse pancreas cancer models in vivo to identify combinations of p53 TSAGs whose loss promotes cancer. We will define the cellular functions of the proteins encoded by p53 TSAGs in different cellular processes regulated by p53, including cell-cycle progression, apoptosis, metabolism, and invasion/metastasis, using overexpression and knockdown approaches. Detailed analyses of p53 TSAG function will help elucidate cellular functions most critical for tumor suppression. We will next use mass spectrometry and siRNA screening to identify the co-factors through which the TADs act to induce p53 TSAGs and to suppress pancreatic cancer, providing another level of understanding of tumor suppression. Finally, we will leverage our expertise in studying p53 TADs to define the importance of TADs for p53 GOF activity. We will use a systematic approach to analyze the TADs at the genetic, genomic and biochemical levels to understand how tumor-derived p53 GOF mutants exert their effects. Collectively, these studies will provide crucial insight into how to modulate p53 pathways during therapeutic strategies for cancer.

Public Health Relevance

In the US, approximately 1 in every 2-3 individuals will develop cancer, a disease of uncontrolled cellular proliferation that can be fatal. It is therefore very important to understand the basis of this disease in order to be able to more effectively treat and cure it. This research aims to elucidate how p53, a gene important for preventing cancer, acts, with the ultimate goal of devising better prevention and treatment strategies for cancer.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Unknown (R35)
Project #
5R35CA197591-06
Application #
9959347
Study Section
Special Emphasis Panel (ZCA1)
Program Officer
Johnson, Ronald L
Project Start
2015-08-14
Project End
2022-07-31
Budget Start
2020-08-01
Budget End
2021-07-31
Support Year
6
Fiscal Year
2020
Total Cost
Indirect Cost
Name
Stanford University
Department
Radiation-Diagnostic/Oncology
Type
Schools of Medicine
DUNS #
009214214
City
Stanford
State
CA
Country
United States
Zip Code
94305
Kaiser, Alyssa M; Attardi, Laura D (2018) Deconstructing networks of p53-mediated tumor suppression in vivo. Cell Death Differ 25:93-103
Mello, Stephano S; Attardi, Laura D (2018) Deciphering p53 signaling in tumor suppression. Curr Opin Cell Biol 51:65-72
Calo, Eliezer; Gu, Bo; Bowen, Margot E et al. (2018) Tissue-selective effects of nucleolar stress and rDNA damage in developmental disorders. Nature 554:112-117
Tarangelo, Amy; Magtanong, Leslie; Bieging-Rolett, Kathryn T et al. (2018) p53 Suppresses Metabolic Stress-Induced Ferroptosis in Cancer Cells. Cell Rep 22:569-575
Mello, Stephano S; Valente, Liz J; Raj, Nitin et al. (2017) A p53 Super-tumor Suppressor Reveals a Tumor Suppressive p53-Ptpn14-Yap Axis in Pancreatic Cancer. Cancer Cell 32:460-473.e6
Van Nostrand, Jeanine L; Bowen, Margot E; Vogel, Hannes et al. (2017) The p53 family members have distinct roles during mammalian embryonic development. Cell Death Differ 24:575-579
Mello, Stephano S; Sinow, Carolyn; Raj, Nitin et al. (2017) Neat1 is a p53-inducible lincRNA essential for transformation suppression. Genes Dev 31:1095-1108
Bieging-Rolett, K T; Johnson, T M; Brady, C A et al. (2016) p19(Arf) is required for the cellular response to chronic DNA damage. Oncogene 35:4414-21