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

项目总结 在美国，胰腺癌是一个主要的健康问题，只有7%的患者存活超过5年 以及几乎所有出现转移的患者。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的途径，以帮助临床治疗 疾病。为此，我们建立了一个小鼠模型来删除Ctnd1基因(该基因编码蛋白质 P120catenin或p120ctn)，这是E-钙粘蛋白稳定性所必需的，其缺失会导致增强 Pdx-cre；KrasG12D；p53f/f(KPC)胰腺上皮间充质转化与转移 肿瘤小鼠模型。我们发现kpc-p120ctnKO小鼠显著增加了相对转移负荷。 来控制产仔动物。对这些小鼠的肿瘤细胞进行无偏筛选导致了对 分泌因子甲状旁腺激素样激素(PTHLH)作为一种信号转导途径 以前不为人知的EMT/转移的贡献者。原位移植中甲状旁腺激素释放激素缺失的研究 实验显示显著降低了肿瘤的生长和转移，表明甲状旁腺素是一种致癌物质。 和促转移因子。此外，抗甲状旁腺激素释放激素单抗的治疗减少。 细胞增殖和迁移，显示出潜在的临床益处。最后，我们生成了一个 KPC-PthlhKO小鼠的初步队列，与KPC对照相比，KPC-PthlhKO小鼠的存活率提高，目前 开始在KPC小鼠身上进行抗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，我们假设甲状旁腺素是胰腺肿瘤发生和转移的驱动因素。 这将通过下列相互关联的具体目标来实现：目标1和目标2是界定 甲状旁腺激素释放激素缺失或抑制对胰腺上皮细胞特性和最终转移定植的影响。 在相同的背景下，我们将探索钙信号(AIM)的其他调节因子(Camk2b和MCU)所起的作用 3)在这些过程中发挥作用，并确定参与钙介导的新的下游信号成分 胰腺癌转移。最后，在目标4中，我们将确定甲状旁腺素在协调免疫中的作用。 微环境和免疫治疗联合治疗。 此K99-R00应用程序中建议的培训概述了指导性研究和 职业发展活动，以及我通往独立研究职业生涯的具体战略 胰腺癌。这个奖项将允许我在以下指导下完善现有技能并获得其他技能 我的研究导师斯坦格博士和鲁斯特吉博士，以及一个跨学科咨询委员会。已被占用 科学提案和培训/职业发展计划将共同提供令人信服的基础 我最终成为了一名成功的NIH资助的独立教员。

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.