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

总结 这是对最初资助的 R01 的竞争性更新，重点是剖析膜联蛋白 A2 介导的 胰腺导管腺癌（PDAC）侵袭和转移的机制。 PDAC 是其中之一 恶性疾病死亡的主要原因。 PDAC的预后不良归因于早期发病 转移和缺乏 PDAC 的有效治疗方法。 PDAC的肿瘤微环境（TME）是 据信这是克服治疗 PDAC 挑战的关键。之前，我们证明了酪氨酸 膜联蛋白 A2 的磷酸化对于 PDAC 侵袭和转移至关重要。我们进一步发现 膜联蛋白 A2 控制信号蛋白 3D (Sema3D) 的分泌以及 Sema3D 对 PDAC 的自分泌作用 通过与其受体 Plexin D1/Neuropilin-1 (PlxnD1/NrP1) 结合来侵袭和转移。有趣的是， Sema3D、PlxnD1 和 NrP1 均属于神经轴突引导蛋白家族，该家族是最重要的神经轴突引导蛋白家族之一。 PDAC 中经常改变的基因家族。 PDAC 中 TME 的神经元成分不能 可以忽略不计，因为神经周围浸润（PNI）是许多恶性肿瘤的重要病理特征， 特别是PDAC。信号蛋白和丛蛋白也被认为通过以下方式调节免疫功能： 控制巨噬细胞的分化和运输。积累的证据表明 巨噬细胞促进腺泡向导管化生（ADM）的发展，这被认为是 PDAC 启动的机制。因此，我们建议检验以下假设： 肿瘤细胞与神经之间的相互作用在促进ADM、生长、侵袭、 和 PDAC 的转移。首先，这个新项目将剖析旁分泌 Sema3D-PlxnD1 信号通路 介导 PNI 和随后的 PDAC 转移。其次，该项目将研究神经源性 Sema3D 通过突变 Kras 肿瘤细胞上的 PlxnD1 受体控制 PDAC 发育 依赖方式。它将检查 Sema3D/PlxnD1 突变体 Kras 是否诱导 Warburg 效应 ARF-GTP信号级联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) 将在体内重新编程。 感觉神经。第三，该项目将研究Sema3D的作用和轴突的遗传改变 小鼠体内促癌炎症反应和 PDAC 发育的指导通路 PDAC 模型和人类 PDAC 标本。轴突导向表达之间的关系 胰腺癌前和恶性上皮、胰内神经和免疫细胞中的分子 并将评估巨噬细胞和其他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