Investigating Cellular Communication in the Tumor Microenvironment in Pancreatic Cancer

项目摘要/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年在全球范围内，这主要是由于缺乏有效的治疗方法。成功治疗的一个主要障碍是 PDA中有丰富的纤维化反应，其中包括癌症相关成纤维细胞(CAF)。虽然很多人的胰腺 肿瘤血运减少，内皮细胞调节免疫成熟和侵袭，肿瘤细胞 新陈代谢和转移扩散。在PDA中，Hedgehog(HH)信号以旁分泌方式起作用， 肿瘤细胞产生HH配体，并通过典型的HH受体ptch1向CAF发出信号，导致 下游通路被激活。小鼠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信号在 PDA非常复杂，并且依赖于上下文。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(目标1)，免疫环境中依赖于HH的改变， 和b)通过成纤维细胞-内皮细胞HH依赖的内皮细胞基因表达和功能的变化 串扰(目标2)。我假设HH信号的联合靶向与免疫结合 有效治疗PDA需要有效的反应或血管系统。我们的初步数据显示Slit2， 在抑制PDA的HH通路过程中，其表达水平降低。先前的工作表明， 上皮性SLIT2缺失促进PDA的神经侵袭和转移。然而，我发现Slit2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。在本提案的目标3中，我将 确定成纤维细胞特异性Slit2在胰腺血管生成和肿瘤生长中的作用。总体而言， 拟议的实验将从根本上对细胞内的串扰有新的理解 胰腺TME，包括处理成纤维细胞-免疫和成纤维细胞-内皮细胞通讯 以及胰腺癌中狭缝-ROBO信号的具体研究。这方面的指导阶段 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。我已经发展出 一个额外的顾问小组，包括癌症生物学领域的国际知名领导人(Andrzej博士 内皮生物学(Jason Spence博士)和免疫学(Bethany Moore博士)。加在一起，这 导师团队制定了一项职业发展计划，重点是研究、合作、演讲、 导师、资助人精神和管理技能的进一步发展是成功领导 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.