Interrogate the interaction between tumor cells and nerves in the tumor microenvironment of pancreatic cancer

总结 这是最初资助的R01的竞争性更新，其专注于剖析膜联蛋白A2介导的 胰腺导管腺癌（PDAC）的侵袭和转移机制。PDAC是一个 恶性疾病死亡的主要原因。PDAC的预后差归因于早期发病， 转移和缺乏PDAC的有效治疗。PDAC的肿瘤微环境（TME）是 被认为是克服治疗PDAC挑战的关键。之前，我们发现酪氨酸 膜联蛋白A2的磷酸化是PDAC侵袭和转移所必需的。我们进一步发现， 膜联蛋白A2控制脑信号蛋白3D（Sema3D）的分泌及其对PDAC的自分泌作用 Plexin D1/Neuropilin-1（PlxnD1/NrP1）是通过与Plexin D1/Neuropilin-1受体结合而参与肿瘤的侵袭和转移。有趣的是， Sema3D、PlxnD1和NrP1都属于神经轴突导向蛋白家族，这是神经轴突导向蛋白家族中 PDAC中频繁改变的基因家族。PDAC中TME的神经元成分不能被 由于神经周围浸润（PNI）是许多恶性肿瘤的重要病理特征， 尤其是PDAC。脑信号蛋白和丛蛋白也涉及通过调节免疫功能来调节免疫功能。 控制巨噬细胞的分化和运输。累积的证据表明， 巨噬细胞促进腺泡到导管化生（ADM）的发展，这被认为是一种 引发PDAC的潜在机制。因此，我们建议测试假设， 肿瘤细胞和神经之间的相互作用在促进ADM生长、侵袭 PDAC的转移。首先，这个新项目将剖析旁分泌Sema3D-PlxnD1信号通路 其介导PNI和随后的PDAC转移。其次，该项目将研究神经衍生的 Sema3D通过突变型Kras肿瘤细胞上的PlxnD1受体控制PDAC的发展， 依赖的方式。它将检查由Sema3D/PlxnD1突变体Kras-1诱导的瓦尔堡效应， ARF-GTP信号级联反应会影响体外巨噬细胞的功能以及是否与肿瘤相关 当Sema3D被组织特异性敲除时，巨噬细胞（TAM）将在体内重编程。 感觉神经第三，本项目将研究Sema3D的作用和轴突的遗传改变 在两种小鼠中的促癌炎症反应和PDAC发展中的体内指导途径 PDAC模型和人PDAC标本。轴突导向的表达与 胰腺癌前病变和恶性上皮细胞、胰腺内神经和免疫细胞中的分子 并评估巨噬细胞和其它免疫细胞的数量和分布。 1

Summary This is a competitive renewal of the originally funded R01 that was focused on dissecting Annexin A2 mediated mechanisms for the invasion and metastasis of pancreatic ductal adenocarcinoma (PDAC). PDAC is one of the major causes of death from malignant diseases. The poor prognosis of PDAC is attributed to early onset of metastasis and lack of effective treatments for PDACs. The tumor microenvironment (TME) of PDAC is believed to hold a key for overcoming the challenge of treating PDAC. Previously, we showed that tyrosine phosphorylation of Annexin A2 is essential for PDAC invasion and metastasis. We further discovered that Annexin A2 controls the secretion of Semaphorin 3D (Sema3D) and the autocrine effect of Sema3D on PDAC invasion and metastasis through binding to its receptor, Plexin D1/Neuropilin-1 (PlxnD1/NrP1). Interestingly, Sema3D, PlxnD1, and NrP1 all belong to the nerve axon guidance protein family, which is among the most frequently altered gene families in PDACs. The neuronal component of the TME in PDACs cannot be negligible as perineural invasion (PNI) is an important pathological characteristic in many malignancies, particularly PDAC. Semaphorins and plexins have also been implicated to regulate immune functions by controlling differentiation and trafficking of macrophages. Accumulated evidence has suggested that macrophages promote the development of the acinar to ductal metaplasia (ADM), which is thought to be a mechanism underlying the initiation of PDACs. Therefore, we propose to test the hypothesis that the interaction between the tumor cells and nerves plays an essential role in promoting ADM, growth, invasion, and metastasis of PDAC. First, this new project will dissect a paracrine Sema3D-PlxnD1 signaling pathway that mediates the PNI and subsequent metastasis of PDAC. Second, the project will study the nerve-derived Sema3D in controlling PDAC development through the PlxnD1 receptor on tumor cells in a mutant Kras- dependent manner. It will examine whether the Warburg effect induced by the Sema3D/PlxnD1-mutant Kras- ARF-GTP signaling cascade will affect the function of macrophages in vitro and whether tumor associated macrophages (TAM) will be reprogrammed in vivo when Sema3D is tissue-specifically knocked out from sensory nerves. Third, this project will investigate the role of Sema3D and the genetic alterations of the axon guidance pathway in vivo in pro-cancerous inflammatory response and PDAC development in both mouse models of PDAC and human PDAC specimens. The relationship between the expression of axon guidance molecules in pancreatic premalignant and malignant epithelia, in intra-pancreatic nerves, and in immune cells and the quantity and distribution of macrophages and other immune cells will be assessed. 1