Investigating Cellular Communication in the Tumor Microenvironment in Pancreatic Cancer

项目概要/摘要 胰腺导管腺癌（PDA）预计将成为癌症相关死亡的第二大原因 到2030年，全球范围内，主要是由于缺乏有效的治疗方法。成功治疗的主要障碍是 PDA中存在大量的纤维化反应，其中包括癌症相关成纤维细胞（CAF）。虽然许多胰腺 肿瘤血管化不足，内皮细胞调节免疫成熟和浸润，肿瘤细胞 代谢和转移性播散。在PDA中，Hedgehog（HH）信号以旁分泌方式起作用， 由此肿瘤细胞产生HH配体并通过经典HH受体PTCH 1向CAF发信号，导致 下游通路激活。在小鼠中进行的遗传学和药理学研究相互矛盾 模型，以及令人失望的HH途径抑制剂的临床试验，表明HH信号传导在 PDA是高度复杂和上下文相关的。该提案旨在揭示HH信令的两个关键方面 在PDA中：a）通过成纤维细胞免疫串扰的免疫景观中的HH依赖性改变（Aim 1）， 和B）通过成纤维细胞-内皮细胞介导的内皮细胞基因表达和功能的HH依赖性变化 串扰（目标2）。我假设HH信号传导的联合靶向与免疫调节相结合， 有效治疗PDA将需要血管系统的响应。我们的初步数据表明，狭缝2， 在PDA中HH途径抑制期间，其调节血管生成。以前的研究表明， 上皮细胞Slit 2缺失促进PDA的神经侵袭和转移然而，我发现Slit 2的大部分是 由成纤维细胞产生，而Robo受体在内皮细胞中表达。在本建议的目标3中，我会 确定Slit 2对胰腺血管生成和肿瘤生长的成纤维细胞特异性贡献。总体看 提出的实验将提供一个从根本上新的理解细胞内的串扰 胰腺TME，包括解决成纤维细胞免疫和成纤维细胞内皮通讯， 以及胰腺癌中SLIT-ROBO信号传导的具体研究。这其中的指导阶段 该奖项将由密歇根大学（UM）的艾伦博士和帕斯卡迪马利亚诺博士监督。I have developed 一个额外的咨询小组，包括国际知名的领导人在癌症生物学领域（博士Andrzej Dlugosz）、内皮生物学（Jason Spence博士）和免疫学（Bethany摩尔博士）。结合起来，这 导师团队制定了一个职业发展计划，重点是研究，合作，演示， 导师制，granitarian，和进一步的管理技能所需的成功领导一个 独立研究实验室。

PROJECT SUMMARY/ABSTRACT Pancreatic ductal adenocarcinoma (PDA) is projected to become the 2nd leading cause of cancer-related deaths world-wide by 2030, largely due a lack of effective treatments. A major barrier to successful therapy is the abundant fibrotic reaction in PDA, which includes cancer-associated fibroblasts (CAFs). While many pancreatic tumors are hypovascularized, endothelial cells regulate immune maturation and infiltration, tumor cell metabolism, and metastatic dissemination. In PDA, Hedgehog (HH) signaling functions in a paracrine manner, whereby tumor cells produce HH ligands and signal to CAFs via the canonical HH receptor PTCH1, leading to downstream pathway activation. There have been contradictory genetic and pharmacologic studies in mouse models, as well as disappointing clinical trials with HH pathway inhibitors, indicating the role of HH signaling in PDA is highly complex and context dependent. This proposal aims to uncover two key aspects of HH signaling in PDA: a) HH-dependent alterations in the immune landscape through fibroblast-immune cross talk (Aim 1), and b) HH-dependent changes in endothelial cell gene expression and function through fibroblast-endothelial cross talk (Aim 2). I hypothesize that combined targeting of HH signaling in combination with the immune response or the vasculature will be required to effectively treat PDA. Our preliminary data indicate that Slit2, which regulates angiogenesis, is reduced during HH pathway inhibition in PDA. Previous work showed that epithelial Slit2 deletion promotes neural invasion and metastasis in PDA. However, I find the majority of Slit2 is produced by fibroblasts, while Robo receptors are expressed in endothelial cells. In Aim 3 of this proposal, I will determine the fibroblast-specific contribution of Slit2 to pancreatic angiogenesis and tumor growth. Overall, the proposed experiments will provide a fundamentally new understanding of cellular cross talk within the pancreatic TME, including addressing fibroblast-immune and fibroblast-endothelial communication as well as a specific investigation of SLIT-ROBO signaling in pancreatic cancer. The mentored phase of this award will be overseen by Drs. Allen and Pasca di Magliano at the University of Michigan (UM). I have developed an additional advisory team to include internationally known leaders in the fields of cancer biology (Dr. Andrzej Dlugosz), endothelial biology (Dr. Jason Spence), and immunology (Dr. Bethany Moore). Combined, this mentorship team has set forth a career development plan focused on research, collaboration, presentations, mentorship, grantsmanship, and the furthering of management skills required to successfully lead an independent research laboratory.