Role of Jmjd1a in hypoxia-induced EMT and prostate cancer stem cells

描述(申请人提供)：前列腺癌是美国男性最常见的恶性肿瘤。侵袭性前列腺癌的一个特征是存在神经内分泌分化(NED)损害，即表达NE和干细胞标记物的细胞团。我们发现，HIF与NE特异的转录因子FOXA2合作，诱导了一个转录程序，决定了人类前列腺癌的NED表型。在HIF和FOXA2共同调控的基因中有Jmjd1a，这是一种激活基因表达的组蛋白H3K9去甲基酶。在目标1中，我们将验证这样的假设，即Jmjd1a被HIF和AR招募到Slug和ZEB2的启动子，在那里它触发H3K9去甲基化，并促进HIF介导的Slug和ZEB2的转录，从而诱导EMT程序。芯片和微阵列分析被用来研究Jmjd1a在EMT基因表达中的整体作用。在目的2中，我们将利用原位前列腺癌模型，研究依赖于Jmjd1a的Slug、ZEB2和Aim1基因的表达在低氧诱导的EMT和转移中的生物学作用。目的3研究HIF、Jmjd1a及其靶基因KLF-4在前列腺癌干细胞和正常前列腺干细胞自我更新和分化中的作用。通过验证组蛋白去甲基酶Jmjd1a在EMT和前列腺干细胞中发挥核心作用的假设，我们建议的研究将建立一个新的范例，研究HIF和Jmjd1a在这些与恶性、预后不良和耐药相关的前列腺癌表型中的缺氧依赖作用，从而为前列腺癌提供新的治疗手段。 K99奖将提供将研究从最初的指导阶段转移到独立阶段的方法，该阶段的重点是鉴定和初步表征Jmjd1a调节的蛋白，这些蛋白有助于缺氧条件下的EMT和前列腺癌干细胞，在独立阶段的工作将集中于Jmjd1a和HIF在缺氧驱动的前列腺干细胞中的作用。SBMRI将提供一个极好的环境(详情请参阅设施和其他资源和机构环境部分)，以指导和指导我的研究，并发展我的独立研究。K99大奖和SMBRI将成为我开始学术生涯的杰出基础。在利用现有的尖端技术、系统、程序和设施的同时，我将有独特的机会得到癌症生物学、信号转导、前列腺癌干细胞、基因表达调控和表观遗传学(详见我的职业发展计划)领域的领导者的指导。我有生物化学、分子和细胞生物学的背景，我觉得这个奖项，通过其指导和独立阶段，将使我处于独特的地位，向前迈进，并为新的和新兴的研究做出巨大贡献，这些研究集中在我提议的涉及缺氧、EMT、前列腺干细胞和表观遗传学的研究上。

DESCRIPTION (provided by applicant): Prostate cancer is the most common malignancy in American men. A hallmark of aggressive prostate tumor is the presence of neuroendocrine differentiation (NED) lesions, clusters of cells expressing NE and stem cell markers. We have found that HIF, in cooperation with a NE-specific transcription factor FoxA2, induces a transcriptional program that determines the NED phenotype of human prostate cancer. Among the genes co-regulated by HIF and FoxA2 is Jmjd1a, a histone H3K9 demethylase that activates gene expression. In Aim 1, we will test the hypothesis that Jmjd1a is recruited by HIF and AR to promoters of Slug and ZEB2, where it triggers H3K9 demethylation and facilitates HIF-mediated transcription of Slug and ZEB2 that in turn induces the EMT program. ChIP-on-chip and microarray analyses are proposed to study global role of Jmjd1a for EMT gene expression. In Aim 2, we will investigate the biological role of Jmjd1a-dependent expression of Slug, ZEB2 and genes identified in Aim1 in hypoxia-induced EMT and metastasis using an orthotopic prostate tumor model. In Aim 3, we will study role of HIF, Jmjd1a and their target gene KLF-4 in self-renewal and differentiation of prostate cancer stem cells and normal prostate stem cells. By testing the hypothesis that the histone demethylase Jmjd1a plays a central role in EMT and prostate stem cells, our proposed studies will establish a new paradigm for hypoxia-dependent role of HIF and Jmjd1a in these prostate cancer phenotypes that are associated with malignancy, poor prognosis and resistance to therapy, thereby providing novel therapeutic modalities for prostate cancer. The K99 Award will provide the means to take the research from the initial mentored phase, which is focused on identification and initial characterization of Jmjd1a regulated proteins that contribute to EMT and prostate cancer stem cells under hypoxia to the independent phase where the work will focus on the role of Jmjd1a and HIF in hypoxia driven prostate stem cells. SBMRI will provide an excellent environment (see Facilities and Other Resources and Institutional Environment sections for details) to mentor and guide my research and to develop my independent studies. The K99 Award and SMBRI will serve as an outstanding foundation for the beginning of my academic career. While taking advantage of the cutting edge technologies, systems, programs, and facilities available I will have the distinct opportunity to be mentored by leaders in the fields of cancer biology, signal transduction, prostate cancer stem cells, gene expression regulation and epigenetics (as detailed in my Career Development Plan). With a background in biochemistry, molecular and cell biology, I feel this Award, via both its Mentored and Independent Phases, will uniquely position me to move forward and contribute immensely to new and emerging studies that focus on my proposed research involving hypoxia, EMT, prostate stem cells and epigenetics.