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: 10441551
• 负责人: Jason R. Pitarresi
• 金额: $ 13.73万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10441551
• 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; Development Plans; Disease; Disease Management; E-Cadherin; Early Diagnosis; Epithelial; Epithelial Cells; Esophagus; Event; Faculty; Family; Foundations; Funding; Gene Expression Profiling; Genes; Genetic; Genetically Engineered Mouse; Goals; Grant; Health; Human; Immune; Immune Evasion; Immunotherapeutic agent; Immunotherapy; 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; 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; therapeutic target; 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.

项目概要 胰腺癌是美国的一个主要健康问题，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治疗 检测方法并发现治疗靶向途径，以帮助临床管理 疾病。为此，我们构建了一个小鼠模型来删除 Ctnnd1 基因（该基因编码蛋白质 p120catenin 或 p120ctn），这是 E-CADHERIN 稳定性所必需的，其删除会导致增强 Pdx-cre 中的上皮间质转化 (EMT) 和转移；克拉斯G12D； p53f/f (KPC) 胰腺 癌症小鼠模型。我们发现 KPC-p120ctnKO 小鼠的转移负担相对于 控制同窝动物。对这些小鼠的肿瘤细胞进行公正的筛选，结果鉴定出 通过分泌因子甲状旁腺激素样激素 (PTHLH) 调节钙信号传导 以前未被重视的 EMT/转移贡献者。重要的是，原位移植中的 Pthlh 缺失 实验显示肿瘤生长和转移显着减少，表明 PTHLH 是一种致癌物质 和促转移因子。此外，用抗 PTHLH 单克隆中和抗体治疗可减少 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，相对于 KPC 对照小鼠，其存活率有所提高，目前正在 在 KPC 小鼠中开展抗 PTHLH 抗体的临床前试验，以确定阻断 PTHLH 是否有效 减弱这种疾病。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 2 是定义 Pthlh 缺失或抑制对胰腺上皮细胞特性以及最终转移定植的影响。 在同样的背景下，我们将探讨钙信号传导的其他调节因子（Camk2b 和 Mcu）的作用（Aim 3）在这些过程中发挥作用并识别参与钙介导的新型下游信号传导成分 胰腺癌转移。最后，在目标 4 中，我们将确定 PTHLH 在协调免疫反应中的作用。 微环境并与免疫疗法共同治疗。 K99-R00 应用程序中拟议的培训概述了指导研究和培训的综合计划 职业发展活动，以及我走向独立研究职业之路的具体策略 在胰腺癌中。该奖项将使我能够在以下人士的指导下完善现有技能并获得额外的技能 我的研究导师，博士。 Stanger 和 Rustgi，以及跨学科咨询委员会。采取 科学建议和培训/职业发展计划共同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 资助的教员。

项目成果

Pancreatic plasticity: Unlocking exocrine lineage specification.

胰腺可塑性：解锁外分泌谱系规范。

• DOI: 10.1016/j.stem.2021.05.006
• 发表时间: 2021
• 期刊: Cell stem cell
• 影响因子: 23.9
• 作者: Pitarresi,JasonR;Rustgi,AnilK
• 通讯作者: Rustgi,AnilK

Single-cell analysis of ductal differentiation.

导管分化的单细胞分析。

• DOI: 10.1038/s41551-021-00782-1
• 发表时间: 2021
• 期刊: Nature biomedical engineering
• 影响因子: 28.1
• 作者: Pitarresi,JasonR;Rustgi,AnilK
• 通讯作者: Rustgi,AnilK

Growth of pancreatic cancers with hemizygous chromosomal 17p loss of MYBBP1A can be preferentially targeted by PARP inhibitors.

• DOI: 10.1126/sciadv.abc4517
• 发表时间: 2020-12
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Hsieh A;Pitarresi JR;Lerner J;Donahue G;Hsiehchen D;Rustgi AK;Zaret K
• 通讯作者: Zaret K