Role of Epigenetic Repression of IRF8 in Tumor Progression/Metastasis

描述（由申请人提供）：结直肠癌是美国男性和女性中第二常见的诊断癌症。结直肠癌患者的绝大多数死亡与肝转移的形成有关。根据美国癌症协会，如果癌症没有转移，结直肠癌患者的5年生存率为92%。然而，一旦癌症转移到肝脏，5年生存率下降到7%。转移性结直肠癌没有有效的治疗方法，治愈的前景仍然很差。对细胞凋亡的抵抗是转移性人结直肠癌的标志。在人类中，Fas在正常结肠组织中组成型表达，然而，Fas在结直肠癌中表达减少，并且在转移性结直肠癌中经常观察到Fas表达和功能的完全丧失。肿瘤细胞凋亡的诱导是许多癌症治疗的基础，包括结肠直肠癌治疗。因此，获得对细胞凋亡的抗性是结直肠癌治疗中最重要的挑战之一。我们已经确定干扰素调节因子8（IRF 8）作为一个重要的调节人大肠癌细胞凋亡。此外，我们观察到，在绝大多数转移性人结直肠癌标本中，IRF 8的表达通过IRF 8启动子DNA超甲基化而沉默。此外，IRF 8的异位表达在体外恢复了转移性人结肠癌细胞对Fas介导的凋亡的敏感性，并在体内抑制了异种移植小鼠模型中原发性人结肠癌的生长。基于这些新的观察，我们假设IRF 8是一种自发的结直肠癌转移抑制因子，通过调节肿瘤细胞对凋亡的敏感性发挥作用，转移性结直肠癌细胞使用IRF 8启动子超甲基化来沉默IRF 8表达以获得转移表型。为了验证我们的假设，我们将追求以下两个具体目标：1）阐明IRF 8在人结肠癌细胞中表达的表观遗传调控的分子机制; 2）确定IRF 8在体内自发性结肠癌转移中的作用，并探索基于IRF 8机制的治疗在抑制结直肠癌转移中的潜力。这项研究项目的成功完成不仅将验证我们的假设，即IRF 8是一种新型的自发转移抑制因子，而且还将为开发基于IRF 8机制的治疗方法以靶向致命的转移性人类结直肠癌提供分子基础。公共卫生相关性：本研究将阐明IRF 8在转移性结肠癌中表达的表观遗传调控的分子机制，并探索靶向IRF 8表达与TRAIL治疗相结合干预人结肠癌体内转移的潜力。这项研究的成功完成将为靶向致命的转移性人类结直肠癌提供一种新的策略。

DESCRIPTION (provided by applicant): Colorectal cancer is the second most commonly diagnosed cancer found in men and women in the United States. The vast majority of mortality of colorectal cancer patients is associated with formation of liver metastasis. According to American Cancer Society, the 5-year survival rate of colorectal cancer patients is 92% if the cancers have not metastasized. However, the 5-year survival rate drops to 7% once the cancer has metastasized to the liver. There is no effective therapy for metastatic colorectal cancer and prospects for cure remains poor. Resistance to apoptosis is a hallmark of metastatic human colorectal cancer. In humans, Fas is constitutively expressed in normal colon tissues, However, Fas expression is diminished in colorectal carcinoma, and complete loss of Fas expression and function is frequently observed in metastatic colorectal cancers. Induction of tumor cell apoptosis is the basis of many cancer therapies, including colorectal cancer therapies. Therefore, acquisition of resistance to apoptosis is one of the most significant challenges in colorectal cancer therapy. We have identified Interferon Regulatory Factor 8 (IRF8) as an essential regulator of apoptosis in human colorectal cancer cells. Furthermore, we observed that IRF8 expression is silenced in the vast majority of metastatic human colorectal cancer specimens through the IRF8 promoter DNA hypermethylation. In addition, ectopic expression of IRF8 restored the sensitivity of metastatic human colon carcinoma cells to Fas-mediated apoptosis in vitro and inhibited the growth of primary human colon carcinoma in a xenograft mouse model in vivo. Based on these novel observations, we hypothesize that IRF8 is a spontaneous colorectal cancer metastasis suppressor that functions through regulating tumor cell sensitivity to apoptosis, and metastatic colorectal cancer cells use the IRF8 promoter hypermethylation to silence IRF8 expression to acquire a metastatic phenotype. To test our hypothesis, we will pursue the following two specific aims: 1) to elucidate the molecular mechanisms underlying epigenetic regulation of IRF8 expression in human colon carcinoma cells; and 2) to determine the roles of IRF8 in spontaneous colon carcinoma metastasis in vivo, and to explore the potential of IRF8 mechanism-based therapies in suppression of colorectal carcinoma metastasis. Successful completion of this proposed research project will not only validate our hypothesis that IRF8 is a novel spontaneous metastasis suppressor but will also provide the molecular basis for the development of IRF8 mechanism-based therapies to target the deadly metastatic human colorectal cancer. PUBLIC HEALTH RELEVANCE: This proposed research project will elucidate the molecular mechanisms underlying epigenetic regulation of IRF8 expression in metastatic human colon cancer, and explore the potential of targeting IRF8 expression in combination with TRAIL therapy for the intervention of human colon carcinoma metastasis in vivo. Successful completion of the proposed research will provide a novel strategy for targeting the deadly metastatic human colorectal cancer.