A novel function of TIMP-1

TIMP-1的新功能

• 批准号: 7758318
• 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/7758318
• 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; base; epithelial to mesenchymal transition; human PHEMX protein; in vivo; inhibitor/antagonist; malignant breast neoplasm; matrigel; member; novel; novel strategies; outcome forecast; overexpression; public health relevance; 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）：该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活性。新出现的临床证据表明 TIMP-1 是不良预后的有效预测因子。然而，除了其作为金属蛋白酶抑制剂的经典活性外，TIMP-1 在乳腺上皮细胞中的作用机制尚不完全清楚。 (MP) 活性，越来越多的证据表明 TIMP-1 也可以独立于 MP 抑制而促进细胞存活。在过去的资助期间，我们最终证明 TIMP-1 是一种有效的内在和外在细胞凋亡抑制剂，独立于其 MP 抑制。重要的是，我们最近的研究确定了四跨膜蛋白成员 CD63 是第一个调节整合素的 TIMP-1 结合细胞表面蛋白。 1 生存途径。CD63/整合素信号复合物的 TIMP-1 激活意味着 TIMP-1 对许多细胞过程的潜在不同影响。初步数据表明，TIMP-1 还抑制非恶性乳腺上皮 MCF10A 的形态发生过程中的管腔形成和细胞凋亡。此外，TIMP-1 与 CD63 的相互作用诱导上皮间质转化。通过上调 EMT 主转录因子 Twist（已知可促进癌症进展），TIMP-1 通过其非 MP 抑制性 C 末端结构域与潜在的 pro-MMP-9 结合。 TIMP-1在细胞表面的调节机制，其中细胞周间隙中pro-MMP-9的可用性可能调节TIMP-1/CD63相互作用。初步研究，我们认为TIMP-1/CD63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 (PI3K)，从而在肿瘤进展和转移的早期阶段导致细胞存活、Twist 表达增加和 EMT。为了检验我们的假设，我们建议 (1)研究TIMP-1和CD63调节整合素异二聚体功能的分子机制，(2)研究TIMP-1/CD63介导的细胞内信号调节细胞存活和 EMT 的转导途径，(3) 确定 pro-MMP-9 在调节 TIMP-1/CD63 介导的细胞存活中的作用，以及 (4) 检查 TIMP 的体内作用-1 在乳腺癌进展过程中。所提出的研究的完成将有助于集体努力了解 TIMP-1 在乳腺癌进展过程中的多种功能。该信息也可能有助于开发针对 TIMP-1 信号传导的新方法。活性与其 MP 抑制活性的关系：金属蛋白酶组织抑制剂 (TIMP)-1 是基质金属蛋白酶 (MP) 的天然抑制剂，已被证明可以抑制体外肿瘤细胞侵袭和后期动物模型中的肿瘤进展。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，根据先前报道的这些研究提供的证据，这是一个意想不到的观察结果。包括我们在内的许多研究人员表明，TIMP-1 也可以独立于其 MP 抑制而促进细胞存活，初步数据表明 TIMP-1 还可以诱导乳腺上皮细胞的上皮间质转化 (EMT) 表型。旨在揭示 TIMP-1 调节细胞存活和 EMT 的分子机制，并检查其在乳腺癌进展早期阶段的动物模型中的潜在致癌活性。有助于集体努力了解 TIMP-1 在乳腺癌进展过程中的多种功能，这一信息也可能有助于开发针对 TIMP-1 信号传导活性及其 MP 抑制活性的新方法。