Functional Interrogation of Kdm6a-Dependent Tumor Suppression during Pancreatic Cancer

项目摘要 我们对导致胰腺癌发生和发展的生物学机制的认识有限， 癌症代表了我们对这种疾病理解的一个根本性差距。如果没有这些知识， 合理开发针对这种疾病的有效治疗方法是极其有限的。我的长期目标是 了解驱动胰腺癌发生的遗传机制。为此，我的总体目标是 这一具体的建议是确定KDM 6A的细胞和分子功能， 已被发现作为一个有前途的新的候选胰腺癌肿瘤抑制剂。两个重要 问题仍然没有答案：1）胰腺癌发生的哪个阶段受到胰腺癌细胞的损失的影响。 Kdm 6a的肿瘤抑制活性依赖于其作为组蛋白的典型功能 脱甲基酶，还是它独立于其脱甲基酶功能？这项建议的理由是， 这些问题，是它将产生对胰腺癌生物学的基本见解， 预计将超过结直肠癌和乳腺癌，成为癌症死亡的第二大原因。 美国到2020年基于现有的数据，我的中心假设是Kdm 6a的丢失驱动了细胞增殖， 胰腺癌细胞的转移倾向，在脱甲基酶非依赖性的方式。向我的中央 为了实现我的总体目标，我将追求两个具体目标：目标1）确定 胰腺癌的新型小鼠模型中肿瘤起始、进展和转移的Kdm 6a损失;以及 目的2）探讨Kdm 6a去甲基化酶活性在胰腺癌抑瘤中的作用 细胞对于目标1，我将使用我最近共同开发的基于CRISPR的小鼠模型来识别Kdm 6a。 在胰腺肿瘤发生过程中。对于目标2，我将进行全面的体外和体内试验， 重新表达野生型或脱甲基酶缺陷型Kdm 6a的Kdm 6a缺失胰腺癌细胞系 确定Kdm 6a的脱甲基酶活性是否是其肿瘤抑制功能所必需的。我相信 这项研究战略是创新的，因为它利用独特的方法论方法来解决 围绕胰腺癌生物学的重要问题，否则难以测试。作为 因此，这项建议有望使癌症研究的新途径超越本报告所述的那些途径。 应用程序.本文提出的研究具有重要意义，因为它将战略性地推动和扩大 我们对胰腺癌知识的界限。最终，这些知识有可能告知 下一代精确靶向治疗药物，将减少美国的癌症负担。

PROJECT SUMMARY Our limited knowledge of the biological mechanisms that lead to the initiation and progression of pancreatic cancer represents a fundamental gap in our understanding of this disease. Without this knowledge, our ability to rationally develop effective treatments for this disease is extremely limited. My long-term goal is to understand the genetic mechanisms that drive pancreatic carcinogenesis. To this end, my overall objective for this specific proposal is to determine both the cellular and molecular function of KDM6A, which has recently been uncovered as a promising novel candidate tumor suppressor of pancreatic cancer. Two important questions remain unanswered: 1) which stage(s) of pancreatic carcinogenesis are affected by the loss of Kdm6a, and 2) is Kdm6a's tumor-suppressive activity dependent on its canonical function as a histone demethylase, or is it independent of its demethylase function? The rationale for this proposal, which addresses these questions, is that it will generate fundamental insights into the biology of pancreatic cancer, which is predicted to surpass both colorectal and breast cancer to become the 2nd leading cause of cancer death in the U.S. by 2020. Based on existing data, my central hypothesis is that loss of Kdm6a drives the proliferation and metastatic proclivity of pancreatic cancer cells in a demethylase-independent manner. To address my central hypothesis and accomplish my overall objective, I will pursue two specific aims: Aim 1) Determine the effect of Kdm6a loss on tumor initiation, progression, and metastasis in a novel mouse model of pancreatic cancer; and Aim 2) Uncover the role of Kdm6a demethylase-activity on its tumor-suppressive function in pancreatic cancer cells. For Aim 1, I will use a CRISPR-based mouse model that I recently co-developed to inactivate Kdm6a during pancreatic tumorigenesis. For Aim 2, I will perform comprehensive in vitro and in vivo assays on Kdm6a-null pancreatic cancer cell lines re-expressing either wild type or demethylase-deficient Kdm6a to determine whether the demethylase activity of Kdm6a is required for its tumor-suppressive function. I believe this research strategy is innovative because it leverages unique methodological approaches to address important questions surrounding pancreatic cancer biology that are otherwise prohibitively difficult to test. As a result, this proposal is expected to enable novel avenues of cancer research beyond those described in this application. The research proposed here is significant because it will strategically push forward and expand the boundaries of our knowledge of pancreatic cancer. Ultimately, this knowledge has the potential to inform the next generation of precisely targeted therapeutics that will reduce the burden of cancer in the United States.