Defining the role of CNPY2 in promoting tumor progression through mediation of macrophage.

摘要 肝癌是最常见的癌症类型之一。超过70万人被诊断出患有这种疾病 全世界每年都有癌症。肝癌导致30,000多例新病例和12,000例 美国每年的死亡人数。已发现肝癌与内质网应激/未折叠高度相关 蛋白质反应(UPR)诱导的慢性炎症；然而，其机制尚不清楚。 使用遗传方法，我们发现巨噬细胞中ER蛋白CNPY2的缺失可以防止 致癌物诱导的肝细胞癌，伴随ER应激/UPR信号减少和肿瘤- 肿瘤中的相关巨噬细胞。此外，促炎症细胞因子、IL-6和 枯否细胞/肝巨噬细胞释放肿瘤坏死因子α。我们还观察到CNPY2在 调节内质网应激/UPR和TLR4信号，这两个已知促进细胞因子产生和 巨噬细胞的分化。总之，这些观察表明CNPY2促进了肝脏肿瘤的发生 通过调节巨噬细胞。 在这项提案中，我将致力于解决肝细胞癌领域的几个基本问题：1)什么是 CNPY2转录调控巨噬细胞产生IL-6和IL-23的机制两者都是 参与这一调节的UPR和TLR4通路？2)什么是UPR和TLR4的生化和结构基础 CNPY2在促进巨噬细胞TLR4信号依赖性细胞因子产生中的作用解决晶体结构问题 将有助于针对CNPY2的药物开发。3)CNPY2的潜在调控机制 肿瘤相关巨噬细胞在肝细胞癌中的分化、侵袭和功能4)角色是什么？ CNPY2与肿瘤免疫？靶向CNPY2能提高肝细胞癌的抗肿瘤免疫功能吗？我们的传统 条件性Cnpy2 KO小鼠是解决这些问题的独特模型。成功地执行此操作 这项工作将大大推进这一领域的发展。从长远来看，这项研究可能会为 基于CNPY2抑制剂的合理设计开发一类新的癌症治疗药物 肿瘤相关巨噬细胞。

Abstract Liver cancer is one of the most common types of cancer. More than 700,000 people are diagnosed with this cancer each year throughout the world. Liver cancer is responsible for more than 30,000 new cases and 12,000 deaths a year in the United States. Liver cancer has been found to be highly associated with ER stress/unfolded protein response (UPR)-induced chronic inflammation; however, the mechanisms remain unclear. Using a genetic approach, we have found that deletion of an ER protein, CNPY2, from macrophages prevents carcinogen-induced hepatocellular carcinoma, accompanied by reduced ER stress/UPR signals and tumor- associated macrophages in tumors. Furthermore, CNPY2 is required for pro-inflammatory cytokines, IL6 and TNFα released from Kupffer cells/liver macrophages. We also observed that CNPY2 plays a central role in regulating both ER stress/UPR and TLR4 signaling, two pathways known to promote cytokine production and differentiation of macrophages. Together these observations suggested that CNPY2 promotes liver oncogenesis through regulation of macrophages. In this proposal, I will aim to address several fundamental questions in the field of HCC: 1) what is the mechanism by which CNPY2 transcriptionally regulates production of IL6 and IL23 in macrophages. Are both the UPR and TLR4 pathways involved in this regulation? 2) what is the biochemical and structural basis for the roles of CNPY2 in promoting TLR4 signaling-dependent cytokine production in macrophages. Solving crystal structure of CNPY2 will help drug development against CNPY2. 3) The potential mechanism by which CNPY2 regulates differentiation, infiltration and function of tumor-associated macrophages in HCC. 4) What is the role of CNPY2 in tumor immunity? Does targeting CNPY2 improve anti-tumor immunity in HCC? Our traditional and conditional Cnpy2 KO mice are unique models for addressing these questions. Successful execution of this work will significantly advance the field. In the longer term, this study may lay a strong foundation for the development of a new class of therapeutics for cancer, based on the rational design of CNPY2 inhibitors against tumor-associated macrophages.