Vitamin D3 alters stromal-epithelial inflammatory crosstalk in human prostate

描述（由申请人提供）：维生素D是一种多效性饮食激素，可防止致癌。流行病学和临床前研究表明，维生素D参与前列腺癌的预防，并可能在前列腺癌的治疗中发挥作用。越来越多的证据表明，炎症是前列腺癌的一个危险因素。我们的总体假设是，维生素D3是前列腺中的抗炎剂，这种活性有助于维生素D3预防前列腺癌。抗炎药可以预防前列腺癌的概念并不新鲜。然而，由于严重的副作用，使用考克斯-2-选择性抑制剂的长期预防临床研究是不实际的，并且使用非处方NSAID的研究难以长时间运行。因此，通过维持足够的维生素D状态来减少前列腺炎症和可能的全身炎症是有吸引力的。 这项新提议是基于我们先前发表的关于维生素D在前列腺上皮细胞中抗炎作用的研究，以及PI资助的K22基金“维生素D调节的microRNAs在前列腺癌预防中的意义”的初步结果。microRNA对炎症信号和反馈回路也至关重要。miR表达的总体和特异性变化已在前列腺癌中得到证实。我们在前列腺上皮细胞和临床试验组织中鉴定了几种受维生素D3调节的炎性miR。这项试验中的男性（N=60，在多伦多大学完成）在根治性乳腺癌切除术前给予400 IU、10，000 IU或40，000 IU维生素D3（胆钙化醇）4-6周。 我们的前期工作主要集中在前列腺上皮，然而，前列腺上皮腺体被前列腺基质包围。众所周知，前列腺基质参与前列腺癌，因为基质对上皮增殖、分化具有调节控制，并且是前列腺炎症反应的必要介质。响应于炎症刺激，上皮细胞和基质细胞迅速释放细胞因子，其传播炎症反应并将免疫细胞募集到该区域。为了探究细胞类型之间的相互作用，我们使用原代前列腺基质细胞和上皮细胞开发了一种三维共培养的体外炎症模型。使用这个模型，我们的初步数据表明，不仅前列腺基质细胞产生不同的细胞因子和microRNA比上皮细胞，但维生素D3消除不同的特定细胞因子和microRNA在每种细胞类型。 本提案中的实验将检验前列腺基质和上皮之间的相互作用被维生素D3改变的假设;创造一个增殖减少、分化增加和炎症减少的微环境。我们提出了一个床边到工作台的方法，首先量化基质组织中的关键基因和microRNA，然后在相关的体外模型中评估这些炎症介质。

DESCRIPTION (provided by applicant): Vitamin D is a pleiotropic dietary hormone that protects against carcinogenesis. Epidemiologic and pre-clinical studies show that vitamin D is involved in prostate cancer prevention and may have a role in prostate cancer treatment. There is mounting evidence that inflammation is a risk factor for prostate cancer. Our overall hypothesis is that vitamin D3 is an anti-inflammatory agent in the prostate and this activity contributes to prostate cancer prevention by vitamin D3. The concept that anti-inflammatory agents may prevent prostate cancer is not new. However, long term prevention clinical studies with COX-2- selective inhibitors are not practical due to severe side effects and studies with non-prescription NSAIDS are difficult to run over long periods of time. Therefore, it is appealing to reduce prostatic inflammation, and likely systemic inflammation, by maintaining adequate vitamin D status. This new proposal is based on our previous published studies on the anti-inflammatory actions of vitamin D in prostate epithelial cells and preliminary results of the PI's funded K22 grant "Significance of Vitamin D-Regulated MicroRNAs in Prostate Cancer Prevention". MicroRNAs are also crucial to inflammatory signaling and feedback loops. Global and specific changes in miR expression have been demonstrated in prostate cancer. We identified several inflammatory miRs that were regulated by vitamin D3 in prostate epithelial cells and clinical trial tissue. Men in this trial (N=60, completed at the University of Toronto) wre given 400IU, 10,000IU or 40,000IU of vitamin D3 (cholecalciferol) for 4-6 weeks prior to radical prostatectomy. Our preliminary work focused on prostatic epithelium, however, prostatic epithelial glands are surrounded by prostate stroma. It is well established the prostate stroma is involved in prostate cancer as the stroma has regulatory control over epithelial proliferation, differentiation and is a necessary mediator of the prostatic inflammatory response. In response to inflammatory stimulus, epithelial and stromal cells rapidly release cytokines that propagate the inflammatory response and recruit immune cells to the area. To interrogate the interplay between cell types, we have developed a three-dimensional co-culture in vitro model of inflammation using primary prostatic stromal and epithelial cells. Using this model, our preliminary data show that not only do prostate stromal cells produce different cytokines and microRNAs than epithelial cells, but that vitamin D3 abrogates different specific cytokines and microRNAs in each cell type. The experiments in this proposal will examine the hypothesis that the interplay between prostate stroma and epithelium is altered by vitamin D3; creating a micro-environment of decreased proliferation, increased differentiation and decreased inflammation. We propose a bedside-to-bench approach to first quantify the key genes and microRNAs in stromal tissue then assess these inflammatory mediators in a relevant in vitro model.