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

描述（由申请人提供）：前列腺癌是美国男性最常见的恶性肿瘤。侵袭性前列腺肿瘤的一个标志是存在神经内分泌分化（NED）病变，表达NE和干细胞标记的细胞簇。我们发现HIF与ne特异性转录因子FoxA2合作，诱导了一个转录程序，该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，一种激活基因表达的组蛋白H3K9去甲基化酶。在Aim 1中，我们将验证Jmjd1a被HIF和AR招募到Slug和ZEB2启动子的假设，在那里它触发H3K9去甲基化，促进HIF介导的Slug和ZEB2转录，进而诱导EMT程序。采用ChIP-on-chip和微阵列分析来研究Jmjd1a在EMT基因表达中的全局作用。在Aim 2中，我们将利用原位前列腺肿瘤模型研究jmjd1a依赖Slug、ZEB2和Aim1中鉴定的基因表达在缺氧诱导的EMT和转移中的生物学作用。在Aim 3中，我们将研究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化酶Jmjd1a在EMT和前列腺干细胞中发挥核心作用的假设，我们提出的研究将为HIF和Jmjd1a在这些与恶性、预后不良和治疗抵抗相关的前列腺癌表型中的缺氧依赖作用建立新的范式，从而为前列腺癌提供新的治疗方式。

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.