Inhibiting serine protease-induced prostate cancer progression

抑制丝氨酸蛋白酶诱导的前列腺癌进展

基本信息

批准号：
8498656
负责人：
Evette S Radisky
金额：
$ 33.83万
依托单位：
MAYO CLINIC JACKSONVILLE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-01 至 2018-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8498656
关键词：
Affinity Androgens Animal Model Antibodies Behavior Biological Biological Assay Biological Markers Cancer Cell Growth Cancer Etiology Cancer Patient Cancer Survivor Cause of Death Cell Culture Techniques Cessation of life Clinical Clinical Management Data Disease Drug Targeting Enzymes Extracellular Matrix Future Genetic Transcription Growth Human Indolent Intervention Investigation Learning Malignant Neoplasms Malignant neoplasm of prostate Measures Mediating Metabolic Pathway Metastatic Prostate Cancer Metastatic to Modeling Molecular Molecular Target Mus Neoplasm Metastasis Organ PTGS2 gene Pathway interactions Patients Peptide Hydrolases Phenotype Preclinical Testing Primary Neoplasm Process Prognostic Marker Prostate Prostate specific antigen measurement Prostate-Specific Antigen Prostatectomy Protein Engineering Proteins Proteolysis Proteomics RNA Interference Radical Prostatectomy Reagent Recombinants Recurrence Relapse Reporter Role Serine Protease Site Staining method Stains Structure Testing Therapeutic Therapeutic Intervention Tissues Trypsin Trypsin Inhibitors Up-Regulation advanced disease base cancer cell chemotherapy cohort experience high risk improved in vivo inhibitor/antagonist insight men mesotrypsin mouse model novel outcome forecast polypeptide prognostic promoter prostate cancer cell protein expression prototype public health relevance research study therapeutic target tool treatment strategy tumor growth tumor progression

Affinity Androgens Animal Model Antibodies Behavior Biological Biological Assay Biological Markers Cancer Cell Growth Cancer Etiology Cancer Patient Cancer Survivor Cause of Death Cell Culture Techniques Cessation of life Clinical Clinical Management Data Disease Drug Targeting Enzymes Extracellular Matrix Future Genetic Transcription Growth Human Indolent Intervention Investigation Learning Malignant Neoplasms Malignant neoplasm of prostate Measures Mediating Metabolic Pathway Metastatic Prostate Cancer Metastatic to Modeling Molecular Molecular Target Mus Neoplasm Metastasis Organ PTGS2 gene Pathway interactions Patients Peptide Hydrolases Phenotype Preclinical Testing Primary Neoplasm Process Prognostic Marker Prostate Prostate specific antigen measurement Prostate-Specific Antigen Prostatectomy Protein Engineering Proteins Proteolysis Proteomics RNA Interference Radical Prostatectomy Reagent Recombinants Recurrence Relapse Reporter Role Serine Protease Site Staining method Stains Structure Testing Therapeutic Therapeutic Intervention Tissues Trypsin Trypsin Inhibitors Up-Regulation advanced disease base cancer cell chemotherapy cohort experience high risk improved in vivo inhibitor/antagonist insight men mesotrypsin mouse model novel outcome forecast polypeptide prognostic promoter prostate cancer cell protein expression prototype public health relevance research study therapeutic target tool treatment strategy tumor growth tumor progression

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项目摘要

DESCRIPTION (provided by applicant): We have identified the serine protease PRSS3/mesotrypsin as a molecule that is prognostic for prostate cancer systemic progression following prostatectomy, and that is upregulated in metastatic prostate cancer tissue. We have also found that silencing of mesotrypsin expression in prostate cancer cells inhibits invasiveness in culture and metastasis in an orthotopic mouse model, while treatment of prostate cancer cells with recombinant mesotrypsin stimulates invasive behavior. Our preliminary data suggest that mesotrypsin promotes prostate cancer progression through cleavage of one or more specific substrates in the extracellular matrix (ECM), leading to upregulation of COX-2 and stimulation of invasive behavior. We hypothesize that mesotrypsin may represent both a potential therapeutic target for metastatic prostate cancer and a useful prognostic tissue biomarker of systemic progression. Here, we propose experiments (1) to further define the biological mechanisms by which mesotrypsin promotes prostate cancer progression, (2) to develop potent and selective mesotrypsin inhibitors that will facilitate mechanistic studies in culture and animal models and offer candidates for mesotrypsin-targeted therapeutics, and (3) to evaluate mesotrypsin as a prognostic tissue biomarker. In Aim 1, we will identify proteolytic substrates of mesotrypsin in ECM using a proteomic approach, and test their impact on invasion using 3D culture models. We will also determine the mechanisms by which mesotrypsin regulates COX-2 transcription using promoter-reporter assays and RNA interference approaches. In Aim 2, we will employ structure-guided protein engineering to optimize a polypeptide inhibitor of mesotrypsin, and evaluate the impact of this inhibitor on invasion in culture assays and on tumor growth and metastasis in an orthotopic mouse model. In Aim 3, we will assess mesotrypsin protein staining as a potential prognostic tissue biomarker in a high-risk radical prostatectomy cohort, using a newly developed selective mesotrypsin antibody. We will also determine whether mesotrypsin expression is associated with metastasis to specific organ sites, using a diverse panel of metastatic tissues. The successful completion of these aims will critically improve our understanding of novel molecular mechanisms that underlie prostate cancer progression.

描述（由申请人提供）：我们已经确定了丝氨酸蛋白酶PRSS3/中丙吡啶蛋白酶是一种分子，对前列腺癌症治疗后的预后是预后的，前列腺切除术后癌症是系统性进展，并且在转移性前列腺癌组织中上调。我们还发现，在原位小鼠模型中，在前列腺癌细胞中表达中性丙二酰表达的沉默抑制了培养和转移中的侵入性，而用重组性中性丙烯蛋白酶对前列腺癌细胞进行治疗会刺激侵入性行为。我们的初步数据表明，间替统蛋白酶通过在细胞外基质（ECM）中的一个或多个特异性底物的裂解促进前列腺癌的进展，从而导致COX-2上调和刺激侵入性行为。我们假设中丙蛋白酶可能代表了转移性前列腺癌的潜在治疗靶标，又代表了系统性进展的有用的预后组织生物标志物。在这里，我们提出了实验（1），以进一步定义生物支替蛋白促进前列腺癌的进展，（2）开发有效的和选择性的间替型蛋白酶抑制剂，这些抑制剂将促进培养和动物模型中的机械研究，并为中ry依的候选者提供培训的候选者，以评估候选者（3）以评估（3）的研究。在AIM 1中，我们将使用蛋白质组学方法来鉴定ECM中间替蛋白的蛋白水解底物，并使用3D培养模型测试其对侵袭的影响。我们还将确定使用启动子重孢子测定和RNA干扰方法来调节Cox-2转录的机制。在AIM 2中，我们将采用结构引导的蛋白质工程来优化美索胰蛋白酶的多肽抑制剂，并评估该抑制剂对培养分析中的侵袭以及对正常小鼠模型中肿瘤生长和转移的影响。在AIM 3中，我们将使用新开发的选择性间替依龙蛋白抗体在高风险的自由基前列腺切除术队列中评估间替肾上腺素蛋白染色作为潜在的预后组织生物标志物。我们还将使用多种转移组织面板确定间替型蛋白酶表达是否与特定器官位点的转移相关。这些目标的成功完成将批判性地提高我们对前列腺癌进展的新型分子机制的理解。