Project 2: Fetuin-A in Prostate Cancer

项目 2：胎球蛋白-A 在前列腺癌中的作用

基本信息

批准号：
10693358
负责人：
ROBERT J. MATUSIK
金额：
$ 7.08万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-09-21 至 2026-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10693358
关键词：
3-Dimensional AKT Signaling Pathway Ablation Address Affect African American American Androgens Attenuated Biogenesis Cancer Etiology Cells Cessation of life Data Death Rate Disease Disease Outcome Doctor of Philosophy Enterobacteria phage P1 Cre recombinase Female Freezing Generations Genes Gleason Grade for Prostate Cancer Glycoproteins Growth Human Immunohistochemistry In Vitro Indolent Invaded Knock-out Knockout Mice LNCaP Laboratories Linear Regressions Malignant neoplasm of prostate Mediating Messenger RNA Mitogen-Activated Protein Kinases Modeling Molecular Mus Mutant Strains Mice Neoplasm Metastasis Null Lymphocytes PIK3CG gene PTEN gene Patient-Focused Outcomes Patients Pharmaceutical Preparations Play Process Production Prognosis Prognostic Marker Prostate Prostate Cancer therapy Prostatic Neoplasms Proteins Proto-Oncogene Proteins c-akt Puberty Refractory Regression Analysis Research Personnel Role Serum Signal Pathway Signal Transduction TLR4 gene The Vanderbilt-Ingram Cancer Center at the Vanderbilt University Tissues Transfection Tumor Volume Western Blotting Xenograft procedure alpha-Fetoproteins biomarker identification calcification cancer health disparity cancer initiation cancer prevention castration resistant prostate cancer caucasian American cell growth cell motility exosome experimental study in vivo mRNA Expression male men mouse model mutant nano-string overexpression probasin prostate cancer cell prostate cancer cell line prostate cancer model prostate cancer progression transmission process tumor tumor growth tumor initiation tumor progression tumorigenic uptake

项目摘要

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.

项目摘要：完整项目 2 尽管前列腺癌 (PCa) 的生长主要是由雄激素驱动的，但其中一部分通常会发展为对雄激素消融有抵抗力（也称为去势抵抗性 PCa；CRPC），具有转移潜力。我们实验室的初步数据表明胎球蛋白-A，也称为 α 2-Heremans-Schmid 糖蛋白（AHSG），在 PCa 细胞的生长和“具有摄取能力”的外泌体的产生中。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。我们追踪PCa细胞表达异位胎球蛋白-A，该异位胎球蛋白-A通过TLR4被细胞分泌并摄取，从而介导“具有摄取能力”外泌体的生物发生，通过激活 pAKT/pERK 促进 PCa 生长；此外，我们假设胎球蛋白 A 表达升高可作为 PCa 3 的预后生物标志物。提出了具体目标：目标 1. 确定 AA PCa 组织中胎球蛋白 A 的表达是否相对较高与高加索裔美国人 (CA) PCa 组织的关系以及高胎球蛋白 A 表达是否与高格里森评分 (>6) 和增强的 pAKT 和 pERK 我们将使用胎球蛋白-A 的 mRNA 表达进行分析。使用人类 PCa 的免疫组织化学 (IHC) 分析 NanoString 以及 pAKT/pERK 蛋白水平多变量线性回归分析将用于确定胎球蛋白-A、pAKT、 AA 和 CA 患者 PCa 组织中的 pERK 和 Gleason 评分，以及其他进展参数，例如由于 PCa 的阳性边缘和扩散，预计胎球蛋白-A、pAKT 和 pERK 将高表达。 AA 患者的 PCa 组织中的水平，特别是具有高格里森评分 (>6) 的患者。目标 2. 确定异位胎球蛋白-A 在外泌体生物合成、促进 2-D 和 3-D 生长、运动性和侵袭性中的作用为此，我们将在两种 PCa 细胞系中过度表达并敲除胎球蛋白-A，以确定 PCa 细胞的能力。胎球蛋白-A是否在传递生长的“摄取能力”外泌体的生物发生中发挥因果作用我们希望证明来自胎球蛋白-A 过表达细胞的外泌体会产生信号。促进 PCa 细胞的 2-D、3-D 生长、运动和侵袭，而来自胎球蛋白 A 缺失细胞的外泌体则不会。目标 3. 研究靶向胎球蛋白 A 介导的信号传导对前列腺抑制的功效小鼠中的肿瘤发生和生长为此目的，我们将利用 Pten-null 小鼠模型来确定 PCa。 Pten 缺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 的缺失是否 Pten-/-/胎球蛋白-A-/-双突变小鼠减弱了Pten-null的致瘤作用，我们预计肿瘤会减少。与 Pten-null 胎球蛋白-A+/+ 小鼠相比，双突变小鼠的生长情况意义：迫切需要。以确定可以区分 CRPC 和惰性 PCa 的生物标志物，该提案解决了这一需求并评估胎球蛋白-A 增强 PCa 肿瘤生长的过程。