Dissecting the oncogenic and pro-metastatic roles of PTHLH-mediated calcium signaling in pancreatic cancer

剖析 PTHLH 介导的钙信号传导在胰腺癌中的致癌和促转移作用

• 批准号: 10696155
• 负责人: Jason R. Pitarresi
• 金额: $ 24.9万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10696155
• 关键词: 3-Dimensional; Accounting; Advisory Committees; Antibodies; Attenuated; Automobile Driving; Award; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Calcium Signaling; Cancer Etiology; Cause of Death; Cell Proliferation; Cells; Cessation of life; Clinical; Clinical Management; Code; Cytoplasm; Development Plans; Disease; Disease Management; Early Diagnosis; Epithelial Cells; Epithelium; Esophagus; Event; Faculty; Family; Foundations; Funding; Gene Expression Profiling; Genes; Genetic; Genetically Engineered Mouse; Goals; Grant; Health; Human; Immune; Immune Evasion; Immunotherapeutic agent; Immunotherapy; KPC model; KRASG12D; Ligands; Malignant Neoplasms; Malignant neoplasm of pancreas; Mediating; Mentors; Mitochondria; Mus; Myeloid-derived suppressor cells; Neoplasm Metastasis; Neoplastic Cell Transformation; Null Lymphocytes; Oncogenes; Oncogenic; Organoids; Pancreas; Pancreatic Ductal Adenocarcinoma; Pathogenesis; Pathway interactions; Patients; Phase; Phenotype; Play; Primary Neoplasm; Process; Prognosis; Proteins; Research; Role; Signal Pathway; Signal Transduction; Solid; Squamous cell carcinoma; Stains; Survival Rate; System; Testing; Therapeutically Targetable; Training; Transplantation; Tumor Suppressor Genes; Tumor-infiltrating immune cells; United States National Institutes of Health; Work; calcium uniporter; calcium-dependent protein kinase; career; career development; catenin p120ctn protein; cell motility; cohort; detection method; efficacy evaluation; efficacy testing; epithelial to mesenchymal transition; experimental study; hormone therapy; improved; member; mouse model; neoplastic cell; neutralizing antibody; neutralizing monoclonal antibodies; novel; novel strategies; novel therapeutics; pancreatic ductal adenocarcinoma cell; pancreatic ductal adenocarcinoma model; pancreatic tumorigenesis; parathyroid hormone-related protein; preclinical trial; recruit; research and development; skills; tumor; tumor growth; tumor-immune system interactions

PROJECT SUMMARY Pancreatic cancer is a major health issue in the US, with only 7% of patients surviving beyond 5 years and nearly all patients presenting with metastases. Identifying factors that increase metastasis may aid early detection methods and discover therapeutically targetable pathways to help with the clinical management of this disease. To this end, we have generated a mouse model to delete the Ctnnd1 gene (which codes for the protein p120catenin or p120ctn), which is necessary for E-CADHERIN stability and whose deletion results in enhanced epithelial-to-mesenchymal transition (EMT) and metastasis in the Pdx-cre; KrasG12D; p53f/f (KPC) pancreatic cancer mouse model. We show that KPC-p120ctnKO mice have dramatically enhanced metastatic burden relative to control littermates. An unbiased screen of tumor cells from these mice resulted in the identification of misregulated calcium signaling through the secreted factor Parathyroid Hormone Like Hormone (PTHLH) as a previously unappreciated contributor of EMT/metastasis. Importantly, Pthlh deletion in orthotopic transplantation experiments showed significantly reduced tumor growth and metastasis, suggesting PTHLH as an oncogenic and pro-metastatic factor. Furthermore, treatment with an anti-PTHLH monoclonal neutralizing antibody reduced cell proliferation and migration, demonstrating a potential clinical benefit. Finally, we have generated a preliminary cohort of KPC-PthlhKO mice that have increased survival relative to KPC controls and are currently embarking on a preclinical trial with anti-PTHLH antibody in KPC mice, to determine if blocking PTHLH will attenuate this disease. Thus, we hypothesize that PTHLH is a driver of pancreatic tumorigenesis and metastasis. This will be achieved through the following interrelated Specific Aims: Aims 1 and 2 are to define the effect of Pthlh deletion or inhibition upon pancreatic epithelial cell identity and ultimately, metastatic colonization. In the same context, we will explore the role that other regulators (Camk2b and Mcu) of calcium signaling (Aim 3) play in these processes and identify novel downstream signaling components involved in calcium-mediated pancreatic cancer metastases. Finally, in Aim 4, we will determine the role of PTHLH in orchestrating the immune microenvironment and co-treat with immunotherapy. The proposed training in this K99-R00 application outlines an integrated plan of mentored research and career development activities, as well as a specific strategy for my pathway to an independent research career in pancreatic cancer. This award will allow me to refine existing and gain additional skills with the guidance of my research mentors, Drs. Stanger and Rustgi, as well as an interdisciplinary advisory committee. Taken together, the scientific proposal and training/career development plans will provide a compelling foundation for me to become eventually a successful independent NIH funded faculty member.

项目摘要 胰腺癌在美国是一个主要的健康问题，只有7%的患者存活超过5年。 并且几乎所有患者都表现出转移。识别增加转移的因素可能有助于早期 检测方法和发现治疗靶向途径，以帮助临床管理这一点， 疾病为此，我们建立了一个小鼠模型，删除Ctnnd 1基因（编码蛋白质 p120 catenin或p120 ctn），其对于E-CADHERIN稳定性是必需的，并且其缺失导致增强的 Pdx-cre; KrasG 12 D; p53 f/f（KPC）胰腺癌中的上皮-间质转化（EMT）和转移 癌症小鼠模型。我们发现KPC-p120 ctnKO小鼠的转移负荷相对于 来控制同窝仔对来自这些小鼠的肿瘤细胞进行无偏筛选， 通过分泌因子甲状旁腺激素样激素（PTHLH）作为钙信号传导的失调， 以前未被认识到的EMT/转移的贡献者。重要的是，原位移植中的Pthlh缺失 实验显示显著降低肿瘤生长和转移，表明PTHLH是一种致癌因子， 和促转移因子。此外，用抗PTHLH单克隆中和抗体治疗降低了 细胞增殖和迁移，证明了潜在的临床益处。最后，我们生成了一个 KPC-PthlhKO小鼠的初步队列，其相对于KPC对照具有增加的存活率，并且目前 在KPC小鼠中进行抗PTHLH抗体的临床前试验，以确定阻断PTHLH是否会 减轻这种疾病。因此，我们假设PTHLH是胰腺肿瘤发生和转移的驱动因素。 这将通过以下相互关联的具体目标来实现：目标1和目标2是确定 Pthlh缺失或抑制对胰腺上皮细胞特性和最终转移性定殖的影响。 在同样的背景下，我们将探讨钙信号（Aim）的其他调节剂（Camk 2b和Mcu）的作用， 3)在这些过程中发挥作用，并确定新的下游信号转导成分参与钙介导的 胰腺癌转移最后，在目标4中，我们将确定PTHLH在协调免疫过程中的作用。 微环境和免疫治疗的共同治疗。 本K99-R 00应用程序中的拟议培训概述了指导研究的综合计划， 职业发展活动，以及我通往独立研究生涯的具体策略 在胰腺癌中。这个奖项将使我能够完善现有的和获得额外的技能的指导下， 我的研究导师，斯坦格博士和鲁斯特吉博士，以及一个跨学科的咨询委员会。采取 科学建议和培训/职业发展计划将为以下方面提供令人信服的基础： 我最终成为一名成功的独立NIH资助的教师。