Genetic effectors of TGF-beta induced growth and arrest.

TGF-β 的遗传效应子诱导生长和停滞。

• 批准号: 7036929
• 负责人: BEN H PARK
• 金额: $ 29.49万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-02-01 至 2011-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7036929
• 关键词: RNA interference; biological signal transduction; cell cycle proteins; cell growth regulation; cell line; cell proliferation; cell senescence; chromatin immunoprecipitation; functional /structural genomics; gene expression profiling; human tissue; mammary epithelium; microarray technology; neoplasm /cancer genetics; oncoprotein p21; phosphoprotein phosphatase; transcription factor; transforming growth factors

DESCRIPTION (provided by applicant): Breast Cancer is a leading cause of cancer death in the United States with ~200,000 new cases and ~40,000 deaths each year. A better understanding of the genetic effectors of tumor growth and suppression would enable the identification of new targets for therapy. The aims of this proposal are directed at identifying and validating key components of the TGF-beta signaling pathway that would lead to the development of specific therapies for the treatment of breast cancer. AIM 1: Analyzing the role of cdc25A in TGF-beta mediated growth stimulation of breast epithelial cells. Our initial studies demonstrated that TGF- beta mediated growth stimulation correlated with an increased expression of the cdc25A tyrosine phosphatase gene in cells lacking p21. In this aim we will further analyze the ability of cdc25A to mediate TGF-beta induced cellular proliferation using RNA interference, promoter/mutational/ChIP assays, and p21 cDNA mutational studies, coupled with an analysis of human breast cancers using tissue microarrays. AIM 2: Identifying mediators of TGF-beta induced cellular proliferation. In Aim 2, we hypothesize that there are other integral genes whose upregulation is needed for TGF-beta induced growth stimulation. We will use microarray expression analysis to identify these genes using our previously characterized p21-/- cell lines. In order to validate that these genes are true mediators of TGF-beta stimulated cellular proliferation, RNA interference gene "knock down" experiments and targeted gene ablation via homologous recombination will be performed. AIM 3: Identifying genetic effectors that mediate the growth response to TGF-beta. In this aim, we will identify additional effectors of TGF-beta mediated growth inhibition using a functional genetic screen. We will combine exon trapping with genetic instability to isolate genes whose inactivation confers resistance to TGF-beta mediated growth suppression. Isolated genes will be verified and validated by restoration of the putative gene and correlated with immunohistochemical studies using tissue microarrays.

描述（由申请人提供）：乳腺癌是美国癌症死亡的主要原因，每年约20万例新病例，死亡约40,000例。更好地了解肿瘤生长和抑制的遗传效应子将使您能够鉴定出新的治疗靶标。该提案的目的旨在识别和验证TGF-β信号通路的关键组成部分，这将导致开发特定的乳腺癌治疗疗法。目标1：分析Cdc25a在TGF-β介导的乳腺上皮细胞的生长刺激中的作用。我们的最初研究表明，TGF-β介导的生长刺激与缺乏p21细胞中Cdc25a酪氨酸磷酸酶基因的表达增加相关。在此目标中，我们将进一步分析Cdc25a使用RNA干扰，启动子/突变/芯片测定和p21 cDNA突变研究介导TGF-β诱导细胞增殖的能力，并与组织微阵列分析人类乳腺癌。 AIM 2：鉴定TGF-β诱导的细胞增殖的介体。在AIM 2中，我们假设还有其他积分基因，其上调TGF-β诱导的生长刺激需要上调。我们将使用微阵列表达分析使用我们先前表征的p21 - / - 细胞系来识别这些基因。为了验证这些基因是TGF-β刺激细胞增殖的真正介体，将进行RNA干扰基因“敲低”实验和通过同源重组的靶向基因消融。目标3：确定介导对TGF-β的生长反应的遗传效应子。在此目标中，我们将使用功能性遗传筛选来确定TGF-β介导的生长抑制的其他效应因子。我们将将外显子捕获与遗传不稳定性结合，以分离出灭活的基因，其灭活性赋予了对TGF-β介导的生长抑制的抗性。分离的基因将通过恢复推定的基因进行验证和验证，并使用组织微阵列与免疫组织化学研究相关。