Identification of the growth-promoting PKD/YAP axis as a novel target for the statins in intestinal epithelial cells.

上皮细胞的序贯增殖、谱系特异性分化、迁移和死亡 肠粘膜的细胞是一个受到严格调控的过程，受到广泛的 调节肽、神经递质、生物活性脂和分化信号。其中许多 刺激通过七螺旋G蛋白偶联受体(GPCRs)起作用。尽管他们 最重要的是，涉及的细胞内信号转导机制仍然 不完全理解。蛋白激酶D(PKD)是GPCR信号通路中的一个关键节点 从而了解PKD在肠上皮细胞中的调节和功能 具有强烈的兴趣和潜在的影响。高度保守的河马/YAP/TAZ途径是 作为器官大小、组织再生的关键调节因素也引起了人们的强烈关注， 肿瘤发生与GPCR信号转导。根据我们的初步结果，我们确定了一部小说 PKD和YAP之间的串扰对促进肠上皮细胞起关键作用 扩散。进一步的初步结果表明，FDA批准的他汀类药物发挥了有效的作用 GPCR/PKD诱导的YAP诱导肠道转录和DNA合成的抑制剂 上皮细胞。他汀类药物还抑制了体外肠样形成，并阻止了再生 损伤后的结肠粘膜。因此，我们的中心假设是：1)PKD/YAP/TAZ 信号在促进肠上皮细胞增殖方面起着至关重要的作用，包括 祖细胞/干细胞和他汀类药物通过靶向抑制肠上皮细胞增殖 PKD/YAP/TAZ轴在这些细胞中表达。一个重要的翻译推论是，广泛使用的 他汀类药物为结直肠癌的化学预防提供了一种新的策略 通过抑制促进生长的PKD/YAP/TAZ轴，从而抑制IBD相关的结直肠癌。三 提出了具体目标：具体目标1：确定脂质过氧化作用的机制(S) 他汀类药物通过PKD/YAP轴降低肠道靶向信号的研究 特异目的2：表征他汀类药物抑制血管内皮细胞生长的影响 PKD/YAP/TEAD通路对在体小肠上皮细胞增殖的影响 体外3D培养的祖细胞和Lgr5+肠干细胞及肠样细胞；特异性目的 3：确定他汀类药物敏感的PKD/YAP/TEAD轴在肠上皮细胞中的作用 再生和致癌。我们预计这项提案的结果将提供一个 实施针对PKD/YAP的创新治疗干预的强有力的理由 消化系统增生性疾病中的信号轴，包括结直肠癌和肠易激综合征 在美国退伍军人以及广大美国民众中存在相关的儿童权利公约。

The sequential proliferation, lineage-specific differentiation, migration and death of the epithelial cells of the intestinal mucosa is a tightly regulated process modulated by a broad range of regulatory peptides, neurotransmitters, bioactive lipids and differentiation signals. Many of these stimuli act through heptahelical G protein-coupled receptors (GPCRs). Despite their fundamental importance, the intracellular signal transduction mechanisms involved remain incompletely understood. Protein kinase D (PKD) is emerging as a key node in GPCR signaling and consequently the understanding of PKD regulation and function in intestinal epithelial cells is of intense interest and potential impact. The highly conserved Hippo/YAP/TAZ pathway is also attracting strong attention as a key regulator of organ-size, tissue regeneration, carcinogenesis and GPCR signaling. Based on our preliminary results, we identified a novel crosstalk between PKD and YAP that plays a critical role in promoting intestinal epithelial cell proliferation. Further preliminary results demonstrate that FDA-approved statins act as potent inhibitors of GPCR/PKD-induced YAP-induced transcription and DNA synthesis in intestinal epithelial cells. Statins also abrogated enteroid formation in vitro and prevented regeneration of colon mucosa after injury. Consequently, our central hypotheses are: 1) PKD/YAP/TAZ signaling plays a vital role in promoting the proliferation of intestinal epithelial cells including progenitor/stem cells and 2) statins restrain intestinal epithelial cell proliferation by targeting the PKD/YAP/TAZ axis in these cells. An important translational corollary is that the widely used drugs of the statin family provide a novel strategy in the chemoprevention of colorectal (CRC) and IBD-related CRC through inhibition of the growth-promoting PKD/YAP/TAZ axis. Three Specific Aims are proposed: SPECIFIC AIM 1: Identify the mechanism(s) by which the lipid- lowering drugs of the statin family target signaling through the PKD/YAP axis in intestinal epithelial cells; SPECIFIC AIM 2: Characterize the impact of statin-induced inhibition of the PKD/YAP/TEAD pathway on the proliferation of intestinal epithelial cells in vivo, including progenitor and Lgr5+ intestinal stem cells and enteroids in 3 D cultures in vitro; SPECIFIC AIM 3: Determine the role of the statin-sensitive PKD/YAP/TEAD axis in intestinal epithelial cell regeneration and carcinogenesis. We anticipate that the outcome of this proposal will provide a robust rationale for implementing innovative therapeutic interventions targeting the PKD/YAP signaling axis in proliferative diseases of the digestive system, including CRC and IBD- associated CRC in US veterans as well as in the US population at large.