TIMP3: A Molecular Target of Green Tea Polyphenols

TIMP3：绿茶多酚的分子靶点

• 批准号: 9099803
• 负责人: SANJAY GUPTA
• 金额: $ 20.68万
• 依托单位: CASE WESTERN RESERVE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9099803
• 关键词: Acetylation; Basement membrane; Binding; Biological Assay; Biological Markers; Breast cancer metastasis; Cancer Patient; Catalytic Domain; Cause of Death; Cell Culture Techniques; Cells; ChIP-seq; Clinical; Clinical Trials; Colon Carcinoma; Complex; Consumption; Data; Deacetylation; Development; Disease; Disease-Free Survival; Down-Regulation; Enhancers; Enzymes; Epigallocatechin Gallate; Epigenetic Process; Extracellular Matrix Degradation; Gelatinase A; Genetic; Genomics; Global Change; Goals; HGF gene; Health; Histone Deacetylase Inhibitor; Histones; Home environment; Homologous Gene; Human; In Vitro; Incidence; Kidney; Knowledge; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of brain; Malignant neoplasm of lung; Malignant neoplasm of prostate; Matrix Metalloproteinases; Mediating; Medical; Metastatic Prostate Cancer; Modality; Modeling; Modification; Molecular; Molecular Target; Monitor; Mus; Neoplasm Metastasis; Organ; Patients; Peptide Hydrolases; Phenotype; Polycomb; Pre-Clinical Model; Preventive; Primary Neoplasm; Process; Prostatic Neoplasms; Protein Subunits; Proteins; Recurrence; Repression; Role; SCID Mice; Staging; Supplementation; Surrogate Endpoint; TIMP3 gene; Tissue Inhibitor of Metalloproteinases; Trichostatin A; Tumor Angiogenesis; Tumor Cell Invasion; Tumor Suppressor Proteins; Up-Regulation; Vascular Endothelial Growth Factors; Work; base; bone; cancer cell; clinical investigation; cost; cost effective; histone methylation; histone modification; implantation; improved; in vivo; inhibitor/antagonist; knock-down; metastasis prevention; metastatic process; mortality; neoplastic cell; novel; novel marker; novel strategies; novel therapeutics; overexpression; prevent; promoter; prostate cancer cell; prostate cancer model; protein expression; research study; soft tissue; stoichiometry; survivorship; tumor; tumor growth; tumor progression

 DESCRIPTION (provided by applicant): The overall goals of this proposal is to understand the mechanisms by which green tea polyphenols and its major constituent (-)-epigallocatechin-3-gallate inhibits prostate cancer metastasis. Consumption of green tea polyphenols (GTP) is effective in the development and could delay the progression of prostate cancer at its outset. However, the molecular mechanism(s) by which green tea polyphenols and its major constituent, epigallocatechin-3-gallate (EGCG) suppresses the metastatic process has not been completely understood. Metastasis is a complex multistep process that involves degradation of extracellular matrix through induction of proteolytic enzymes viz. matrix metalloproteinases (MMPs) and downregulation of their binding partner tissue inhibitor of metalloproteinases (TIMPs) critical for invasion and dissemination of malignant cells. Our preliminary studies demonstrate that GTP/EGCG-mediated tumor cell invasion correlate with decrease in MMP-2/9 expression along with concomitant increase in TIMP-3 in tumor cells. Based on our exciting preliminary results we hypothesize that GTP/EGCG-mediated re-expression of TIMP-3 is a critical mechanism wherein GTP/EGCG inhibits acquisition of invasiveness by prostate cancer cells and prevents tumor metastasis. TIMP-3 directly binds to the catalytic domain of matrix metalloproteinase (MMP-2 and -9) in 1:1 stoichiometry, which are key enzymes involved in the dissolution of basement membrane. Low level of TIMP-3 protein expression has been shown to be associated with an aggressive tumor phenotype and poor disease-free survival. Under the proposed specific aim 1 we will elucidate the molecular mechanism(s) by which GTP/EGCG upregulates TIMP-3 and suppress invasiveness in prostate cancer cells. Through experiments outlined in specific aim 2, we plan to determine the in vivo effects of GTP/EGCG in upregulating TIMP-3 and preventing metastasis in preclinical models. The hypothesis is that in vivo supplementation of GTP/EGCG could effectively increase TIMP-3 expression through global modification in histone proteins and specific alterations at the TIMP-3 promoter significantly reduces metastatic incidence in pre-clinical models. We plan to use an orthotopic implantation tumor model in castrated nu/nu mice which forms primary tumor in the organ and metastasizes to the bone and an intracardiac model of tumor metastasis using SCID mice which home tumors to various soft tissue. These studies will provide mechanistic basis and novel biomarkers for clinical investigation of GTP/EGCG in the prevention of metastasis at early stages thereby reducing the mortality and increase survivorship in prostate cancer patients suffering with recurrent form of the disease.

 描述（由申请人提供）：本提案的总体目标是了解绿色茶多酚及其主要成分（-）-表没食子儿茶素-3-没食子酸酯抑制前列腺癌转移的机制。食用绿色茶多酚（GTP）对前列腺癌的发展是有效的，可以在前列腺癌开始时延缓其进展。然而，绿色茶多酚及其主要成分表没食子儿茶素-3-没食子酸酯（EGCG）抑制转移过程的分子机制尚未完全了解。转移是一个复杂的多步骤过程，涉及通过诱导蛋白水解酶即基质金属蛋白酶（MMP）降解细胞外基质，并下调其结合伴侣金属蛋白酶组织抑制剂（TIMP），其对于恶性细胞的侵袭和扩散至关重要。我们的初步研究表明，GTP/EGCG介导的肿瘤细胞侵袭与MMP-2/9表达的降低沿着TIMP-3在肿瘤细胞中的伴随增加相关。基于我们令人兴奋的初步结果，我们假设GTP/EGCG介导的TIMP-3的再表达是GTP/EGCG抑制前列腺癌细胞获得侵袭力并防止肿瘤转移的关键机制。TIMP-3与基质金属蛋白酶（MMP-2和MMP-9）的催化结构域以1：1的化学计量比直接结合，MMP-2和MMP-9是参与基底膜溶解的关键酶。TIMP-3蛋白表达水平低与肿瘤侵袭性表型和无病生存率低有关。在提出的具体目标1下，我们将阐明GTP/EGCG上调TIMP-3并抑制前列腺癌细胞侵袭的分子机制。通过具体目标2中概述的实验，我们计划确定GTP/EGCG在临床前模型中上调TIMP-3和预防转移的体内作用。该假设是，在体内补充GTP/EGCG可以通过组蛋白的整体修饰有效地增加TIMP-3的表达，并且TIMP-3启动子的特异性改变显著降低了临床前模型中的转移发生率。我们计划在去势nu/nu小鼠中使用原位植入肿瘤模型，其在器官中形成原发性肿瘤并转移到骨，以及使用SCID小鼠的心内肿瘤转移模型，其将肿瘤转移到各种软组织。这些研究将为临床研究GTP/EGCG在早期阶段预防转移提供机制基础和新的生物标志物，从而降低患有疾病复发形式的前列腺癌患者的死亡率并增加存活率。