TBEL Project 2

TBEL项目2

• 批准号: 10518938
• 负责人: ANIRBAN MAITRA
• 金额: $ 39.2万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-21 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10518938
• 关键词: 3-Dimensional; Abdomen; Accounting; Affect; Alleles; Biochemical; Biogenesis; Cell Line; Cell Survival; Clinic; Collaborations; Colorectal Cancer; Cyst; Cystic Lesion; Data; Dependence; Diagnosis; Drops; Excision; Exhibits; Fibroblasts; Frequencies; Genetic; Genetic Engineering; Histopathology; Human; Image; In Vitro; Indolent; Intervention; KRAS2 gene; Lesion; Location; Malignant Neoplasms; Malignant neoplasm of pancreas; Membrane Potentials; Metabolic Control; Metabolic Pathway; Metabolism; Mitochondria; Modeling; Molecular; Molecular Biology Techniques; Morphology; Mucinous Neoplasm; Mus; Mutate; Mutation; Nature; Operative Surgical Procedures; Organ; Organelles; Organoids; Oxidative Phosphorylation; Pancreas; Pancreatic Cyst; Pancreatic Ductal Adenocarcinoma; Pancreatic cystic neoplasia; Papillary; Pathogenesis; Patients; Pharmacology; Phase I Clinical Trials; Pre-Clinical Model; Prevalence; Probability; Property; Proteins; Quality Control; Recording of previous events; Reporter; Reporting; Respiration; Role; Site; Solid; TP53 gene; Testing; Tissues; WNT Signaling Pathway; Wood material; base; cancer invasiveness; clinical application; conditional knockout; endoplasmic reticulum stress; functional loss; genetic approach; genetically modified cells; high risk; improved; in vitro Model; in vivo; inhibitor; insight; mitochondrial metabolism; mouse model; mutant; neoplastic; novel; pancreatic ductal adenocarcinoma model; proteostasis; reconstruction; response; tool; tumor; tumor microenvironment; tumor progression

PROJECT 2 – ABSTRACT The estimated prevalence of pancreatic cystic neoplasms (PCNs) is between 20–30% with intraductal papillary mucinous neoplasm (IPMNs) accounting for half of them. Depending on the morphology, location, and genetics IPMNs can be of high-risk developing pancreatic ductal adenocarcinoma (PDAC) and as such significantly diminishing patient’s survival. While RNF43 is only mutated in a small fraction of PDAC, RNF43 mutations are prevalent in a high frequency in IPMNs. Although RNF43 has been identified as a negative regulator of Wnt signaling in colorectal cancers and other preclinical models including PDAC, a phase 1 clinical trial of a Wnt inhibitor in patients with RNF43 mutated solid cancers (including PDAC) has been proven disappointing. Other likely organ specific functions of RNF43 might be of great advantage for IPMN progression. Utilizing a conditional knockout mouse model of RNF43 in context of Kras mutation we found that RNF43 abrogates the mitochondrial properties and functions while loss of RNF43 improved mitochondrial quality control, increased unfolded protein response (UPR) and ER stress. These findings let us hypothesize that loss of RNF43 may deregulate the crosstalk of these organelles in order to maintain proteostasis, metabolic control, and cell survival. Employing an autochthonous mouse model for functional loss of RNF43 in the context of Kras (generated in the Dr. Maitra lab), primary genetically engineered cell lines (generated in the Dr. Lyssiotis lab) and human IPMN derived organoids (generated in the Dr. Wood lab; see Project 3) we propose to pursue the following aims: First we will investigate whether RNF43 regulates mitochondrial dynamics including biogenesis, fission, fusion and mitophagy to inhibit IPMN pathogenesis (Aim 1). Further, we will investigate whether RNF43 blocks IPMN pathogenesis by limiting ER stress through management of ER-mitochondrial dynamics (Aim 2). Lastly, we will determine whether loss of RNF43 creates a synthetic essentiality for sustained OXPHOS (Aim 1) and whether/how OXPHOS inhibition impacts the multistep progression in an autochthonous KRC model of pancreatic cystic neoplasia (Aim 3). Overall, these studies will elucidate the functional role of RNF43 in mitochondria quality control, ER-mitochondrial dynamics and how this impacts IPMN pathogenesis and progression.

项目 2 – 摘要 导管内乳头状胰腺囊性肿瘤 (PCN) 的估计患病率在 20-30% 之间 其中粘液性肿瘤（IPMN）占一半。取决于形态、位置和遗传学 IPMN 可能是高风险的胰腺导管腺癌 (PDAC)，因此显着 降低患者的生存率。虽然 RNF43 仅在 PDAC 的一小部分中发生突变，但 RNF43 突变 在 IPMN 中出现频率较高。尽管RNF43已被确定为Wnt的负调节因子 结直肠癌和其他临床前模型中的信号转导，包括 PDAC（Wnt 的 1 期临床试验） 事实证明，RNF43 突变实体癌（包括 PDAC）患者的抑制剂效果令人失望。其他 RNF43 可能具有的器官特异性功能可能对 IPMN 进展具有很大优势。使用条件式 在 Kras 突变背景下的 RNF43 敲除小鼠模型中，我们发现 RNF43 消除了线粒体 特性和功能，而 RNF43 的缺失改善了线粒体质量控制，增加了未折叠蛋白 反应（UPR）和 ER 应激。这些发现让我们假设 RNF43 的缺失可能会放松对 这些细胞器的串扰以维持蛋白质稳态、代谢控制和细胞存活。 采用本地小鼠模型来研究 Kras 背景下 RNF43 功能丧失的情况（在 Maitra 博士实验室）、原代基因工程细胞系（在 Lyssiotis 博士实验室生成）和人类 IPMN 衍生类器官（在 Wood 博士实验室生成；参见项目 3），我们建议实现以下目标： 首先我们将研究 RNF43 是否调节线粒体动力学，包括生物发生、裂变、融合 和线粒体自噬抑制 IPMN 发病机制（目标 1）。此外，我们将研究 RNF43 是否阻断 IPMN 通过管理内质网线粒体动力学来限制内质网应激（目标 2）。最后，我们将 确定 RNF43 的缺失是否会产生持续 OXPHOS 的合成必要性（目标 1）以及 OXPHOS 抑制是否/如何影响本地 KRC 模型中的多步进展 胰腺囊性肿瘤（目标 3）。总的来说，这些研究将阐明 RNF43 在 线粒体质量控制、ER 线粒体动力学及其如何影响 IPMN 发病机制和 进展。