A novel function of TIMP-1

TIMP-1的新功能

• 批准号: 7591225
• 负责人: Hyeong-Reh Choi Kim
• 金额: $ 26.94万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-07-01 至 2013-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7591225
• 关键词: 1-Phosphatidylinositol 3-Kinase; Acinus organ component; Animal Model; Apoptosis; Apoptotic; Binding; Breast; C-terminal; Cancer Patient; Cell Surface Proteins; Cell Survival; Cell physiology; Cell surface; Clinical; Complex; Data; Development; Diagnostic Neoplasm Staging; Epithelial; Epithelial Cells; Focal Adhesions; Funding; Goals; Human; In Vitro; Integrins; Light; MCF10A cells; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Mediating; Mesenchymal; Metalloproteases; Molecular; Morphogenesis; Neoplasm Metastasis; Non-Malignant; Oncogenic; Outcome; PTK2 gene; Pathway interactions; Phenotype; Regulation; Reporting; Research Personnel; Role; Signal Transduction; Signal Transduction Pathway; Staging; Testing; Time; Tissue Inhibitor of Metalloproteinase-1; Tissue Inhibitor of Metalloproteinases; Tumor Cell Invasion; Tumor stage; Up-Regulation; abstracting; base; epithelial to mesenchymal transition; human PHEMX protein; in vivo; inhibitor/antagonist; malignant breast neoplasm; matrigel; member; novel; novel strategies; outcome forecast; overexpression; transcription factor; tumor progression

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to unveil the pleiotropic activity of tissue inhibitor of metalloproteinases (TIMP)-1 during breast cancer progression. Emerging clinical evidence indicates that TIMP-1 is a potent predictor of poor prognosis in breast cancer patients. However, the mechanisms of action of TIMP-1 in breast epithelial cells are not completely understood. In addition to its classical activity as an inhibitor of metalloproteinase (MP) activity, accumulating evidence indicates that TIMP-1 can also promote cell survival independently of MP inhibition. During the past funding period, we conclusively showed that TIMP-1 is a potent inhibitor of both intrinsic and extrinsic apoptosis independent of its MP inhibitory activity. Importantly, our recent study identified the tetraspanin member CD63 as the first TIMP-1 binding cell surface protein which modulates the integrin ¿1 survival pathway. TIMP-1 activation of the CD63/integrin signaling complex implies potentially diverse effects of TIMP-1 on many cellular processes. Preliminary data show that TIMP-1 also inhibits lumen formation and apoptosis during morphogenesis of non-malignant breast epithelial MCF10A acini in three-dimensional matrigel cultures. Moreover, TIMP-1 interaction with CD63 induces an epithelial mesenchymal transition (EMT) via upregulation of the EMT master transcription factor Twist, known to promote cancer progression. Structurally, TIMP-1 binds to CD63 via its non-MP inhibitory C-terminal domain as it does to latent pro-MMP-9. This suggests a novel mechanism of TIMP-1 regulation at the cell surface, in which the availability of pro-MMP-9 in the pericellular space may regulate TIMP-1/CD63 interactions. Based on our previous and preliminary studies, we hypothesize that TIMP-1/CD63 activation of integrin ¿1/a signaling complex activates focal adhesion (FAK) and phosphatidylinositol 3-kinase (PI3K) leading to cell survival, increased Twist expression and EMT during the early stages of tumor progression and metastasis. To test our hypothesis, we propose (1) to investigate the molecular mechanisms by which TIMP-1 and CD63 regulate integrin heterodimer functions, (2) to investigate TIMP-1/CD63-mediated intracellular signal transduction pathways for the regulation of cell survival and EMT, (3) to establish the role of pro-MMP-9 in the regulation of TIMP-1/CD63-mediated cell survival, and (4) to examine the in vivo roles of TIMP-1 during breast cancer progression. Accomplishment of the proposed studies will contribute to the collective endeavor to understand the multi functions of TIMP-1 during breast cancer progression. This information may also be useful for the development of novel approaches for targeting TIMP-1's signaling activity versus its MP inhibitory activity. PUBLIC HEALTH RELEVANCE: Tissue inhibitor of metalloproteinases (TIMP)-1, a natural inhibitor of matrix metalloproteinases (MPs), has been shown to inhibit tumor cell invasion in vitro and tumor progression in animal models of later stages of tumor metastasis. However, emerging clinical evidence indicates that TIMP-1 is a potent predictor of poor prognosis in breast cancer patients, an unexpected observation in light of these previously reported studies. Accumulating evidence provided by many investigators including us indicates that TIMP-1 can also promote cell survival independently of its MP inhibition. Interestingly, the preliminary data show that TIMP-1 also induces epithelial mesenchymal transition (EMT) phenotype in breast epithelial cells. The goals of the current application are to unveil molecular mechanisms by which TIMP-1 regulates cell survival and EMT, and to examine its potential oncogenic activity in an animal model of the early stages of breast cancer progression. Accomplishment of the proposed studies will contribute to the collective endeavor to understand the multiple functions of TIMP-1 during breast cancer progression. This information may also be useful for the development of novel approaches for targeting TIMP-1's signaling activity versus its MP inhibitory activity.

描述（由申请人提供）：本提案的长期目标是揭示金属蛋白酶组织抑制剂（TIMP）-1在乳腺癌进展过程中的多效性活性。 新出现的临床证据表明，TIMP-1是乳腺癌患者预后不良的一个有效预测因子。 然而，TIMP-1在乳腺上皮细胞中的作用机制还不完全清楚。 除了其作为金属蛋白酶（MP）活性抑制剂的经典活性之外，越来越多的证据表明TIMP-1还可以独立于MP抑制而促进细胞存活。 在过去的资助期间，我们最终表明，TIMP-1是一种有效的抑制剂，内源性和外源性细胞凋亡的独立的MP抑制活性。 重要的是，我们最近的研究确定了四跨膜蛋白成员CD 63作为第一个TIMP-1结合细胞表面蛋白，它调节整合素1的生存途径。 TIMP-1对CD 63/整联蛋白信号传导复合物的激活暗示TIMP-1对许多细胞过程的潜在不同作用。 初步数据显示，TIMP-1还抑制三维基质胶培养物中非恶性乳腺上皮MCF 10A腺泡形态发生过程中的管腔形成和细胞凋亡。 此外，TIMP-1与CD 63的相互作用通过上调EMT主转录因子Twist诱导上皮间质转化（EMT），已知其促进癌症进展。 在结构上，TIMP-1通过其非MP抑制性C-末端结构域与CD 63结合，就像它与潜在的pro-MMP-9结合一样。 这表明TIMP-1在细胞表面调节的新机制，其中细胞周围空间中MMP-9原的可用性可调节TIMP-1/CD 63相互作用。 基于我们以前的和初步的研究，我们假设TIMP-1/CD 63激活整合素1/a信号复合物激活粘着斑（FAK）和磷脂酰肌醇3-激酶（PI 3 K），导致细胞存活，增加Twist表达和EMT在肿瘤进展和转移的早期阶段。 为了验证我们的假设，我们提出（1）研究TIMP-1和CD 63调节整合素异源二聚体功能的分子机制，（2）研究TIMP-1/CD 63介导的细胞内信号转导途径对细胞存活和EMT的调节，（3）建立pro-MMP-9在TIMP-1/CD 63介导的细胞存活调节中的作用，以及（4）检测TIMP-1在乳腺癌进展过程中的体内作用。 这些研究的完成将有助于理解TIMP-1在乳腺癌进展过程中的多功能。 该信息也可用于开发靶向TIMP-1的信号传导活性与其MP抑制活性的新方法。 公共卫生关系：金属蛋白酶组织抑制剂（TIMP）-1是一种天然的基质金属蛋白酶（MP）抑制剂，在体外可抑制肿瘤细胞的侵袭，在晚期肿瘤转移的动物模型中可抑制肿瘤的进展。 然而，新出现的临床证据表明，TIMP-1是乳腺癌患者预后不良的有效预测因子，根据这些先前报道的研究，这是一个意想不到的观察结果。 包括我们在内的许多研究者提供的越来越多的证据表明，TIMP-1也可以独立于其MP抑制促进细胞存活。 有趣的是，初步数据显示TIMP-1也诱导乳腺上皮细胞中的上皮间质转化（EMT）表型。 本申请的目的是揭示TIMP-1调节细胞存活和EMT的分子机制，并在乳腺癌进展早期的动物模型中检查其潜在的致癌活性。 完成拟议的研究将有助于集体奋进了解TIMP-1在乳腺癌进展过程中的多种功能。 该信息也可用于开发靶向TIMP-1的信号传导活性与其MP抑制活性的新方法。