Investigating HIF-MYC Interactions within a Clear Cell Renal Cell Carcinoma Murine Model

项目摘要/摘要 肾细胞癌(RCC)在男性最常见的癌症中排名第7和第10位 据估计，2015年新增病例61 500例，死亡14 000例。 明确地说，肾透明细胞癌(CcRCC)约占肾脏的70%。 成人中的恶性肿瘤。 已经进行了广泛的研究，以了解潜在的基因变化 慢性肾细胞癌的病理特点。早期和最近的研究表明，大多数 在慢性肾细胞癌患者中发生的常见突变是肿瘤抑制基因的失活， VHL。这些发现导致了针对以下目标的靶向疗法的产生 血管内皮生长因子(或其受体)和mTOR。然而，原发肾细胞癌对这些药物的反应 都是可变的。因此，需要发现靶向的替代和有效的分子靶标 药物治疗是必不可少的。 我们的团队已经建立了一种新的ccRCC基因工程小鼠模型(GEMM)， 与慢性肾细胞癌患者分享组织学。我将利用这个模型作为一个平台来研究分子 缺氧诱导因子α亚基与原癌基因myc的相互作用。我假设HIF1α和 Hif2α(H1H2)和Hif2α(H2)表达对肾小管上皮细胞分化调控的研究 基因组中的转录活性和染色质定位。此外，我 假设MYC的过度表达增加了HIFα亚基的稳定性 尽管失去了VHL。利用我们的GEMM ccRCC模型和NEXT生成的细胞系 代序列分析(RNA-SEQ和CHIP-SEQ)，我将着手询问以下问题 目的：1)研究MYC的转录活性在ccRCC之间是否存在差异 表达H1H2的细胞与仅表达H2的细胞相比，2)MYC如何正向调节HIF1α和 CcRCC内HIF2α水平。

Project Summary/Abstract Renal cell carcinoma (RCC) ranks among the 7th and 10th most common cancer among men and women respectively, with an estimated 61,500 new cases and 14,000 deaths in 2015. Specifically, clear-cell renal cell carcinoma (ccRCC) accounts for approximately 70% of renal malignancies among adults. Extensive research has been conducted on understanding the genetic alterations underlying the pathological features of ccRCC. Early and more recent studies have shown that the most common mutation to occur within ccRCC patients is the inactivation of the tumor suppressor gene, VHL. Such discoveries have led to the generation of targeted therapies that are directed toward VEGF (or its receptor) and mTOR. However, responses of primary ccRCC tumors to theses agents are variable. As such, the need to discover alternative and efficient molecular targets for targeted drug therapy is essential. Our group has generated a novel genetically engineered murine model (GEMM) of ccRCC that shares histology to ccRCC patients. I will utilize this model as a platform to investigate the molecular interactions between HIFα subunits and the proto-oncogene, MYC. I hypothesize that HIF1α and HIF2α (H1H2) and HIF2α (H2) expressing ccRCC cells differentially regulate MYC's transcriptional activity and chromatin localization throughout the genome. Additionally, I hypothesize that the overexpression of MYC confers additional stability to HIFα subunits despite the loss of Vhl. Utilizing cell lines generated from our GEMM ccRCC model and Next Generation Sequencing analysis (RNA-seq and ChIP-seq), I will set out to interrogate the following aims: 1) Investigate whether MYC's transcriptional activity is differentially altered between ccRCC cells expressing H1H2 versus H2 only and 2) Determine how MYC positively regulates HIF1α and HIF2α levels within ccRCC.!