The CCK-B receptor signaling pathway as a driver of pancreatic cellular plasticity and carcinogenesis

摘要 人们已经充分认识到，慢性胰腺炎引起的炎症与8倍的风险增加有关 导致胰腺癌的发生当细胞受到损伤或压力时，它们可以转化为具有 通过一个叫做化生的过程改变表型。胰腺腺泡导管化生（ADM） 发生胰腺炎，这个过程通常是可逆的。然而，随着慢性炎症或成熟 当细胞停止生长时，导管表型细胞不逆转为它们的正常腺泡表型，但可能进展为胰腺癌。 上皮内瘤变（PanIN）和胰腺癌。我们对这些机制的理解存在差距 参与恢复或丧失恢复能力。我们实验室一直在研究G蛋白偶联受体 并发现胆囊收缩素B受体（CCK-BR），发现在导管细胞，而不是腺泡细胞， 在慢性胰腺炎和PanIN形成期间表达，并且在胰腺癌中显著过表达。 癌用CCK受体拮抗剂丙谷胺治疗，加速ADM的恢复，减少ADM的分泌， 炎症，并使胰腺恢复正常表型。在患有PanIN或 胰腺癌，丙谷胺治疗也改变胰腺细胞外基质或肿瘤 微环境，通过减少胰腺星状细胞或癌症相关细胞的胶原蛋白产生， 成纤维细胞和改变免疫细胞的签名，以更正常的表型。我们假设 CCK-BR信号通路是胰腺可塑性的一条新的关键通路， 这个途径会减少胰腺癌的发生。丙谷胺已经在人类受试者中进行了测试， 1期临床试验，被认为具有广泛的安全性特征。在本建议中，我们将研究 丙谷胺和CCK-BR在胰腺细胞可塑性中的作用，以及如何靶向这一途径以使胰腺细胞可塑性正常化。 胰腺表型并使其致癌性降低以预防胰腺癌。以下目标是 提出：目标#1，确定CCK-BR信号通路如何参与腺泡-导管化生;目标 #2，检查CCK-B受体途径的激活如何调节胰腺星状细胞，如果CCK- 受体阻断可以改变表型，使成纤维细胞致癌性降低;目标3，评价 CCK-BR的激活如何诱导组织炎症和免疫细胞的极化。在这次调查中， 我们将使用野生型和CCK-BR基因敲除小鼠的胰腺炎小鼠模型， ADM和基质的激光显微切割、反相蛋白阵列、质谱分析和 RNA测序以了解胰腺细胞的可塑性。我们还将研究CCK-BR在以下方面的作用 胰腺星状细胞在体外使用鼠和人类细胞。我们的目标是了解丙谷胺 将化生或发育不良的胰腺重新编程为更正常的表型， 胰腺癌

Abstract It has become well recognized that inflammation from chronic pancreatitis is associated an 8-fold increased risk for the development of pancreatic cancer. When cells are injured or stressed, they can transform to a cell with a different phenotype through a process called metaplasia. In the pancreas, acinar-ductal metaplasia (ADM) occurs with pancreatitis and this process is usually reversible. However, with chronic inflammation or maturation arrest, the ductal phenotype cells do not reverse to their normal acinar phenotype but may progress to pancreatic intraepithelial neoplasia (PanINs) and pancreatic cancer. There is a gap in our understanding of the mechanisms involved in this recovery or loss of the ability to recover. Our lab has been studying G-protein coupled receptors and found that the cholecystokinin-B receptor (CCK-BR) that is found in ductal cells but not acinar cells becomes expressed during chronic pancreatitis and PanIN formation, and is markedly over-expressed in pancreatic cancer. Treatment with a CCK-receptor antagonist, proglumide, hastens recovery of ADM, decreases inflammation, and restores the pancreas to a normal phenotype. In more advanced disease with PanINs or with pancreatic cancer, proglumide treatment also alters the pancreas extracellular matrix or tumor microenvironment, by decreasing collagen production from pancreatic stellate cells or cancer-associated fibroblasts and changing the immune cell signature to a more normal phenotype. We hypothesize that the CCK-BR signaling pathway is a novel and key pathway in pancreatic plasticity; strategies to suppress this pathway will decrease pancreatic cancer. Proglumide has already been tested in human subjects in a Phase 1 clinical trial and deemed to have a broad safety profile. In this proposal we will study the role of proglumide and the CCK-BR in pancreatic cell plasticity and how this pathway can be targeted to normalize the pancreatic phenotype and render it less oncogenic to prevent pancreatic cancer. The following aims are proposed: Aim #1, Determine how the CCK-BR signaling pathway is involved in acinar-ductal metaplasia; Aim #2, Examine how activation of the CCK-B receptor pathway modulates pancreatic stellate cells and if CCK- receptor blockade can change the phenotype to render the fibroblasts less carcinogenic; and Aim #3, Evaluate how activation of the CCK-BR induces tissue inflammation and polarization of immune cells. In this investigation, we will use murine models of pancreatitis in wild-type and in CCK-BR-knockout mice with state of the art techniques as laser microdissection of ADM and stroma, Reverse phase protein array, Mass Cytometry and RNA sequencing to understand plasticity of pancreatic cells. We will also examine the role of the CCK-BR on the pancreatic stellate cells in vitro using murine and human cells. Our goal is to understand how proglumide reprograms a metaplastic or dysplastic pancreas to a more normal phenotype and thus preventing development of pancreatic cancer.