Defining the mechanism of inositol 1,4,5-triphosphate receptor-mediated metastatic liver colonization in colorectal cancer

项目概要/摘要 转移性进展是实体器官肿瘤如结直肠癌死亡的主要原因， 在分子水平上知之甚少。虽然现代系统治疗已经逐步改善了结果， 转移性结直肠癌的存活率仍然很低，只有不到15%的患者存活5年。 因此，迫切需要鉴定和表征促进细胞凋亡的关键基因和途径。 转移性定植使用基因组规模的体内短发夹RNA（shRNA）筛选，PI和他的 合作者已经鉴定了几种假定的转移促进剂，包括3型肌醇1，4，5- 三磷酸盐受体（ITPR 3），其从内质网动员细胞内钙。ITPR3 在结肠直肠癌细胞中的缺失降低了体内肝转移能力，暗示ITPR 3是结肠直肠癌的关键驱动因素。 转移初步数据表明，ITPR 3可能允许扩散的癌细胞克服生理学上的细胞毒性。 转移性微环境固有的屏障，如正常细胞外附着的丧失， 初级上皮部位，可能通过关闭能量需求过程，如蛋白质 翻译.此外，ITPR 3可以调节促转移的非经典NF-κ B信号传导和相关的细胞凋亡。 转录因子RELB，其也在shRNA筛选中被鉴定为稳健的转移启动子。这些 研究提出了ITPR 3是支持生存的多种途径的中心协调者的假设 在早期和晚期阶段的转移定植。实验提出（1）定义的机制 ITPR 3通过多聚核糖体/核糖体分析、蛋白质组学和生物信息学来调节翻译和细胞代谢 （2）阐明ITPR 3调控RELB和非经典NF-κ B的机制 （3）确定ITPR 3是否可以使用新的治疗靶向， 来源于高转移性患者来源的异种移植物的类器官。圆满完成拟议工作 将提高对结直肠癌转移定植机制的理解，并揭示 开发更有效的治疗策略以阻断转移进展的途径。申请人博士 Ryan Moy是纪念医院的肿瘤内科研究员和胃肠肿瘤科成员 斯隆凯特琳癌症中心（MSKCC）。Moy博士概述了一个5年职业发展计划， 他在分子生物学和肿瘤学临床培训的背景，以获得成为一个重要的技能， 独立的物理学家和科学家他将在Sohail博士的指导下进行拟议的研究 Tavazoie是一位物理学家兼科学家，也是转移生物学领域的领先专家，拥有良好的培训记录 成功的科学家Moy博士将在生物信息学，代谢组学，蛋白质组学和生物信息学方面发展新的技能。 患者来源的癌症模型的生成。除了这个训练计划和强大的科学背景， MSKCC和洛克菲勒大学的综合机构环境使Moy博士能够 成功发展成为一个独立实验室为基础的医生，科学家与R 01的资金。

PROJECT SUMMARY/ABSTRACT Metastatic progression is the predominant cause of death from solid organ tumors like colorectal cancer but is poorly understood at the molecular level. While modern systemic therapy has incrementally improved outcomes, survival in metastatic colorectal cancer remains dismal, with less than 15 percent of patients alive at five years. Therefore, there is an urgent need to identify and characterize the key genes and pathways that facilitate metastatic colonization. Using a genome-scale in vivo short hairpin RNA (shRNA) screen, the PI and his collaborators have identified several putative metastasis promoters including the type 3 inositol 1,4,5- triphosphate receptor (ITPR3), which mobilizes intracellular calcium from the endoplasmic reticulum. ITPR3 deletion in colorectal cancer cells reduces liver metastatic capacity in vivo, implicating ITPR3 as a key driver of metastasis. Preliminary data suggest that ITPR3 may allow disseminating cancer cells to overcome physiological barriers inherent to the metastatic microenvironment, such as the loss of normal extracellular attachment to the primary epithelial site, potentially through shutting down energetically demanding processes such as protein translation. In addition, ITPR3 may regulate pro-metastatic non-canonical NF-kB signaling and the associated transcription factor RELB, which was also identified as a robust metastasis promoter in the shRNA screen. These studies have led to the hypothesis that ITPR3 is a central coordinator of multiple pathways to support survival during early and late phases of metastatic colonization. Experiments are proposed to (1) define the mechanisms by which ITPR3 regulates translation and cellular metabolism through polysome/ribosome profiling, proteomics and metabolomics, (2) elucidate the mechanism by which ITPR3 controls RELB and non-canonical NF-kB signaling to facilitate metastasis, and (3) determine whether ITPR3 can be therapeutically targeted using novel organoids derived from highly metastatic patient-derived xenografts. Successful completion of the proposed work will improve understanding of the mechanisms underlying metastatic colonization in colorectal cancer and reveal avenues to develop more effective therapeutic strategies to block metastatic progression. The applicant, Dr. Ryan Moy, is a Medical Oncology Fellow and member of the Gastrointestinal Oncology Service at Memorial Sloan Kettering Cancer Center (MSKCC). Dr. Moy has outlined a 5-year career development plan that expands on his background in molecular biology and clinical training in oncology to acquire vital skills for becoming an independent physician-scientist. He will perform the proposed studies under the mentorship of Dr. Sohail Tavazoie, a physician-scientist and leading expert in metastasis biology with a strong track record of training successful scientists. Dr. Moy will develop new skills in bioinformatics, metabolomics, proteomics and the generation of patient-derived cancer models. In addition to this training plan and strong scientific background, the combined institutional environments of MSKCC and The Rockefeller University ideally position Dr. Moy to successful develop into an independent laboratory-based physician-scientist with R01 funding.