Project 2 Fetuin-A in Prostate Cancer

项目 2 胎球蛋白-A 在前列腺癌中的应用

• 批准号: 10705093
• 负责人: JOSIAH OCHIENG
• 金额: $ 22.57万
• 依托单位: MEHARRY MEDICAL COLLEGE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-26 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705093
• 关键词: 3-Dimensional; AKT Signaling Pathway; Ablation; Address; Affect; African American; American; Androgens; Attenuated; Biogenesis; Cancer Etiology; Caucasians; 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; Liver; 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; mortality; mouse model; mutant; nano-string; nanovesicle; overexpression; particle; 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

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。这 拟议研究的目的是确定胎球蛋白-A 在前列腺癌进展中的作用和意义。 我们假设 PCa 细胞表达异位胎球蛋白-A，该细胞通过 TLR4 分泌并摄取 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 的预后生物标志物。三 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。我们将使用以下方法分析胎球蛋白-A 的 mRNA 表达 使用人类 PCa 的免疫组织化学 (IHC) 分析 NanoString 以及 pAKT/pERK 蛋白水平 组织。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 作为正利润率和 PCa 的传播。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，特别是那些格里森评分高 (>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 PCa 细胞的容量。为此，我们将在两种 PCa 细胞系中过表达并敲除胎球蛋白-A，以确定 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 受体细胞中的信号。我们期望证明来自胎球蛋白-A 过表达细胞的外泌体将 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 小鼠肿瘤的发生和生长。为此，我们将利用 PCa 的 Pten-null 小鼠模型来确定 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-/-/fetuin-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。