Project 2 Fetuin-A in Prostate Cancer

项目总结：完整项目2 尽管前列腺癌(PCa)的生长在很大程度上是由雄激素驱动的，但通常会出现一个亚群 对雄激素消融(也称为抗去势PCa；CRPC)不耐受，具有转移的潜力。 我们实验室的初步数据表明胎球蛋白-A，也被称为α2-Heremans-Schmid. 糖蛋白(AHSG)在前列腺癌细胞的生长和产生“有摄取能力的”外切体中起重要作用。这个 本研究的目的是明确胎球蛋白-A在前列腺癌进展中的作用和意义。 我们假设前列腺癌细胞表达异位的胎球蛋白-A，它是由细胞通过TLR4分泌和摄取的 通过PACK/PERK的激活，介导具有摄取能力的外体的生物发生，从而促进PCA的生长； 此外，我们假设胎球蛋白-A的表达升高可作为PCa的预后生物标志物。三 目的：1.确定胎球蛋白-A在AA和PCa组织中的表达是否相对较高 高加索美国人(CA)的PCa组织以及胎球蛋白-A高表达是否与高 格里森得分(&gt；6)和增强的帕克特和福利。我们将分析胎球蛋白-A的mRNA表达 应用免疫组织化学(IHC)分析人前列腺癌的NanoStringand PakT/PERK蛋白水平 纸巾。将使用多元线性回归分析来确定胎球蛋白-A、PACT、 AA和CA患者PCa组织中的PERK和Gleason评分以及其他进展参数 作为PCa的正边缘和扩散。预计胎球蛋白-A、PARKT和PERK将在高水平表达 AA患者前列腺癌组织中的水平，尤其是那些Gleason评分高的患者(&gt；6)。目标2.确定 异位胎球蛋白-A在外体生物发生、促进二维和三维生长、运动和侵袭中的作用 PCa细胞的容量。为此，我们将在两个PCa细胞系中过表达并敲除Fetuin-A，以确定 胎球蛋白-A在传递生长的“有摄取能力的”外体的生物发生中是否起因果作用 受体细胞中的信号。我们希望证明，来自胎球蛋白-A过度表达细胞的外切体将 促进PCa细胞的2-D、3-D生长、运动和侵袭，而来自胎蛋白-A缺失细胞的外体则不会。 目的3.研究靶向胎球蛋白-A介导的信号转导对前列腺抑制的作用 肿瘤在小鼠体内的形成和生长。为此，我们将利用Pten-Null小鼠模型来确定PCA Pten丢失是否需要完整的胎球蛋白-A基因来介导其致瘤作用，以及胎球蛋白-A的丢失是否在 PTEN-/-/Fetuin-A-/-双突变小鼠减弱Pten-Null的致瘤作用。我们预计肿瘤会减少 与Pten缺失的胎球蛋白-A/小鼠相比，双突变小鼠的生长发育。重要意义：有迫切的需要 识别可以区分CRPC和惰性PCA的生物标志物，这项建议解决了需要和 评估胎球蛋白-A促进PCa肿瘤生长的过程。

PROJECT SUMMARY: FULL PROJECT 2 Even though the growth of prostate cancer (PCa) is largely driven by androgens, a subset usually develops that is refractory to androgen ablation (also known as castration resistant PCa; CRPC) with potential for metastasis. Preliminary data from our laboratory has implicated fetuin-A, also known as alpha 2-Heremans-Schmid glycoprotein (AHSG), in the growth of PCa cells and in the production of “uptake-competent” exosomes. The objective of the proposed studies is to define the role and significance of fetuin-A in prostate cancer progression. We hypothesize that PCa cells express ectopic fetuin-A which is secreted and taken up by the cells via TLR4 to mediate the biogenesis of `uptake-competent' exosomes that promote PCa growth via activation of pAKT/pERK; moreover, we postulate that elevated fetuin-A expression serves as a prognostic biomarker for PCa. Three specific aims are proposed: Aim 1. To determine if fetuin-A expression is higher in AA PCa tissues relative to Caucasian American (CA) PCa tissues and whether high fetuin-A expression is associated with high Gleason Scores (>6) and enhanced pAKT and pERK. We will analyze mRNA expression of fetuin-A using NanoString as well as pAKT/pERK protein levels using immunohistochemistry (IHC) analysis of human PCa tissues. Multivariable linear regression analysis will be used to determine the correlation between fetuin-A, pAKT, pERK and Gleason scores in PCa tissues of AA and CA patients, as well as other progression parameters such as positive margins and spread of PCa. It is expected that fetuin-A, pAKT and pERK will be expressed at high levels in PCa tissues of AA patients particularly those with high Gleason scores (>6). Aim 2. To determine the role of ectopic fetuin-A in exosome biogenesis, promotion of 2-D and 3-D growth, motility and invasive capacity of PCa cells. In this aim, we will overexpress and knockout fetuin-A in two PCa cell lines to determine whether fetuin-A plays a causal role in the biogenesis of `uptake competent' exosomes that transmit growth signals in recipient cells. We expect to demonstrate that exosomes from fetuin-A overexpressing cells will promote 2-D, 3-D growth and motility and invasion of PCa cells while exosomes from fetuin-A null cells will not. Aim 3. To investigate the efficacy of targeting fetuin-A mediated signaling on the suppression of prostate tumor initiation and growth in mice. In this aim, we will utilize the Pten-null mouse model for PCa to determine whether Pten loss requires intact fetuin-A gene to mediate its tumorigenic role and whether loss of fetuin-A in Pten-/-/fetuin-A-/- double mutant mice attenuates the tumorigenic role of Pten-null. We expect reduced tumor growth in the double mutant mice compared to Pten-null fetuin-A+/+ mice. Significance: There is an urgent need to identify biomarkers that can differentiate CRPC from indolent PCa and this proposal addresses that need and evaluates the process by which fetuin-A enhances PCa tumor growth.