Abstract 2331: Deletion of ΔNp63 and ΔNp73 in p53 deficient mice results in TAp63 and TAp73 compensation of p53 tumor suppression in vivo.

Abstract

Abstract p53 tumor suppressor undergoes mutational loss in majority of cancers contributing to tumor formation. Therapeutic strategies are aimed towards p53 overexpression in tumors or to identify targets that compensate for p53-functional loss. p63 & p73, share structural similarities to p53, making them excellent candidates for therapeutic compensation of p53. Unlike p53, p63 and p73 do not undergo mutational loss and their role in tumorigenesis is being delineated. p63 and p73 have two major isoforms, the transactivation (TA), with activities similar to p53 and the delta (Δ)N- isoform with oncogenic functions. Inhibition of TAp63 and TAp73 is observed in cancers as a consequence of overexpression of ΔN isoforms of p63 and p73. In disparity, recent studies report, tumor suppressive properties of ΔNp63 and ΔNp73 in activating genes involved in DNA repair and apoptosis. To define the functional roles of ΔNp63 and ΔNp73 in cancer, mouse models targeting the ΔN isoforms were generated. We observed that, ΔNp63+/- and ΔNp73−/− mice on a p53−/− background had lower thymic lymphoma incidence compared to the p53−/− mice. I found TAp63 and TAp73 up regulated in the double mutant mice that correspond with an increase in p53-downstream apoptotic (PUMA, Noxa, BAX) and cell cycle targets (p21, p16, PML). This suggests that ablation of ΔN isoforms mediate TAp63 and TAp73 up regulation inducing apoptosis or cell cycle arrest by activation of p53-downstream targets. To further demonstrate this, I ablated ΔNp63 and ΔNp73 in vivo in p53−/- mice thymic lymphoma by administering adenoviral-CRE specifically to the thymus. The CRE-treated mice had a significant thymic lymphoma regression within 3 weeks as imaged by MRI in comparison to the mock-treated mouse cohorts. Additionally, RNA-Seq analysis from CRE-treated versus untreated mice, has identified novel metabolic genes with apoptotic or cell-cycle functions. We further report, ΔNp63 and ΔNp73 to bind to promoter site of TAp63 and TAp73 by chromatin immunoprecipitation (ChIP). This supports the notion that ablation of ΔN isoforms of p63 and p73 restores the function of TAp63 and TAp73 thus compensating for p53-tumor suppressive function in vivo. To test, if ablation of ΔN isoforms reduces tumorigenesis in human cancers, ΔNp63 and ΔNp73 were knocked down in human cancer cell lines were p53 expression was ablated or mutated. TAp63 and TAp73 were upregulated in ΔNp63/ΔNp73 knock down human cancer cell lines. However, induction of apoptosis or cell-cycle arrest was observed in p53-deleted cancer cell lines in comparison to the p53-mutated cell lines. This highlights the co-repressive effect of mutant p53, preventing activation of TAp63/TAp73 downstream targets. Current work is aimed towards overcoming mutant p53 effect in these cancer cell lines. Thus, targeting the ΔNp63/ΔNp73 compensates for p53-functional loss mediating tumor suppression. Citation Format: Avinashnarayan Venkatanarayan, Deepavali Chakravarti, Xiaohua Su, Santosh Sandur, Lingzhi Liu, Eliot Fletcher Sananikone, Payal Raulji, Cristian Coarfa, William Norton, Preethi Gunaratne, Elsa Renee Flores. Deletion of ΔNp63 and ΔNp73 in p53 deficient mice results in TAp63 and TAp73 compensation of p53 tumor suppression in vivo. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2331. doi:10.1158/1538-7445.AM2013-2331