FUNCTIONAL AND IMMUNOPHENOTYPIC ANALYSIS OF P53 AND RB

P53 和 RB 的功能和免疫表型分析

• 批准号: 6424526
• 负责人: CARLOS CORDON-CARDO
• 金额: $ 23.06万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-03-01 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6424526
• 关键词: DNA binding protein; DNA damage; adult human (21+); apoptosis; cell cycle proteins; cell growth regulation; clinical research; cyclin dependent kinase; cyclins; gene expression; gene mutation; genetic transduction; human genetic material tag; human subject; human tissue; metastasis; neoplasm /cancer genetics; oncoprotein p21; p53 gene /protein; prognosis; protein structure function; retinoblastoma protein; sarcoma; transcription factor; tumor suppressor genes

The objective of the proposed studies is the continued characterization of mutations and aberrant patterns of expression of cell cycle regulators as they rate to tumorigenesis and tumor progression in adult soft tissue sarcomas (STS). Our working hypothesis has been that mutations and altered expression of the TP53 and RB genes produce a selective advantage for tumor growth and aggressive behavior in STS patients. Their critical role requires stringent multi-level regulation by other factors such as cyclins, cyclin-dependent kinases (Cdk), and cyclin-dependent kinase inhibitors (Cki). Studies of these regulators are a logical extension of our previous work. The main goal of our project is to translate basic and clinical research findings into clinical studies. The Specific Aims are outlined as follows: Aim #1. Prognostic Significance of TP53 and RB in Adult Soft Tissue Sarcomas: Molecular and Functional Analyses. This aim is divided into two studies. In Study A we plan to: 1) prospectively validate the prognostic value of TP53 and RB; 2) determine the functional significance of different TP53 mutations; 3) determine the molecular basis of altered pRB expression and relating E2F levels with response to chemotherapy. In Study B we will determine if particular STS subtypes have specific molecular "signatures" with respect to p53 and pRB. Aim #2. Molecular Characterization of the p53-Pathway: Impact on Cell Cycle Arrest and Apoptosis. We will determine: 1) the clinical relevance of altered patterns of p21 expression; 2) the proportion of STS that over- express mdm2 by lack MDM2 amplification; 3) the proportion of STS cases that over-express mdm2 but do not express p19/ARF; and 4) the potential clinical impact of joint p53 and mdm2 alterations. Aim #3. Molecular Characterization of the RB-Pathway: Impact on S- Phase Entry and Proliferation. We will determine: 1) the frequency, clinical significance, and nature of pRB over-expression; 2) the mutation frequency of INK4A in STS, and if these mutations involve p16, p19/ARF, or both; 3) the proportion of cases that over-express cyclin D1 and Cdk4, and what proportion is due to gene amplifications; 4) the frequency of co-amplification of CDK4 and MDM2; and 5 ) the frequency and potential clinical relevance of detecting altered expression patterns of E2F proteins (mainly E2F1, E2F4, and E2F5).

拟议研究的目的是持续表征细胞周期调节因子的突变和异常表达模式，因为它们与成人软组织肉瘤（STS）的肿瘤发生和肿瘤进展有关。我们的工作假设是，TP53 和 RB 基因的突变和表达改变为 STS 患者的肿瘤生长和攻击行为产生了选择性优势。它们的关键作用需要其他因素的严格多级调节，例如细胞周期蛋白、细胞周期蛋白依赖性激酶（Cdk）和细胞周期蛋白依赖性激酶抑制剂（Cki）。对这些监管机构的研究是我们之前工作的逻辑延伸。我们项目的主要目标是将基础和临床研究成果转化为临床研究。具体目标概述如下： 目标#1。 TP53 和 RB 在成人软组织肉瘤中的预后意义：分子和功能分析。这一目标分为两项研究。在研究 A 中，我们计划：1）前瞻性验证 TP53 和 RB 的预后价值； 2）确定不同TP53突变的功能意义； 3)确定pRB表达改变的分子基础以及将E2F水平与化疗反应相关联。在研究 B 中，我们将确定特定的 STS 亚型是否具有 p53 和 pRB 的特定分子“特征”。目标#2。 p53 通路的分子表征：对细胞周期停滞和细胞凋亡的影响。我们将确定：1）p21 表达模式改变的临床相关性； 2) 由于缺乏MDM2扩增而过度表达mdm2的STS的比例； 3) 过度表达 mdm2 但不表达 p19/ARF 的 STS 病例比例； 4) p53 和 mdm2 联合改变的潜在临床影响。目标#3。 RB 通路的分子表征：对 S 相进入和增殖的影响。我们将确定：1）pRB 过度表达的频率、临床意义和性质； 2) STS中INK4A的突变频率，以及这些突变是否涉及p16、p19/ARF或两者； 3）cyclin D1和Cdk4过度表达的病例比例，以及由于基因扩增导致的比例； 4) CDK4和MDM2共扩增的频率； 5) 检测 E2F 蛋白（主要是 E2F1、E2F4 和 E2F5）表达模式改变的频率和潜在临床相关性。