Abstract B5: Identification of a role for REST in promoting preneoplastic events in a novel genetically engineered mouse model

Abstract

Medulloblastoma is a pediatric cerebellar tumor that arises from undifferentiated neuronal stem/progenitor cells (NSCs). The Repressor Element-1 (RE1) Silencing Transcription Factor (REST) negatively regulates neurogenesis by repressing numerous genes involved in terminal neuronal differentiation. We previously showed that REST levels were abnormally elevated in medulloblastoma samples and that this was associated with poor prognosis for patients. Previous studies from two groups including ours showed that REST knockdown in human tumor cells blocked their tumorigenic potential in mouse orthotopic models, suggesting that REST was required for tumor maintenance. In addition, constitutively elevated expression of REST in neural progenitors in the context of Myc overexpression promoted tumorigenesis in the murine brain, implicating REST in tumor progression. However, a role for REST in tumor initiation has not been demonstrated. To examine if elevated REST expression in neuronal progenitors was sufficient to promote pre-neoplastic events such as proliferation, delayed differentiation and genetic instability, we generated a novel transgenic (Tg) mouse model in which human REST Tg expression could be conditionally elevated in the post-natal cerebellar progenitor cells. These studies revealed that cerebellar progenitors harvested from REST Tg mice, had sustained neurosphere formation and proliferation potential compared to progenitors from age-matched wild type littermates. Progenitors from REST Tg mice also exhibited substantially decreased expression of REST target neuronal differentiation genes such as Syn1 and SCG10 and displayed delayed onset of neurogenesis compared to wild type controls. Interestingly, progenitors with elevated REST expression had significantly perturbed telomere homeostasis associated with altered telomerase levels and increased genomic instability compared to wild type controls. The above findings were recapitulated in vivo where cerebella from REST Tg mice exhibited prolonged Ki-67 staining accompanied by lack of differentiation and accumulation of genetic instability. Furthermore, REST mediated genetic instability was associated with increased sensitivity to DNA damaging reagents such as etoposide compared to normal wild type progenitors. Thus, we provide the first demonstration of a role for elevated REST expression in neural progenitors in promoting tumor initiating events such as hyperproliferation, blockade of differentiation and telomere defects. We also show that the genetic instability contributes to the accelerated response of these cells to clinically relevant DNA damage-inducing agents. These observations may have therapeutic implications for high REST medulloblastomas. Citation Format: Tara Dobson, Ellen Busschers, Pete Taylor, Julianne Humphries, Sadhan Majumder, Vidya Gopalakrishnan. Identification of a role for REST in promoting preneoplastic events in a novel genetically engineered mouse model. [abstract]. In: Proceedings of the AACR Special Conference on Pediatric Cancer at the Crossroads: Translating Discovery into Improved Outcomes; Nov 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;74(20 Suppl):Abstract nr B5.