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：
9020758
负责人：
Evette S Radisky
金额：
$ 32.53万
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/9020758
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 Cessation of life Clinical Clinical Management Data Disease Drug Targeting Enzymes Extracellular Matrix Future Genetic Transcription Growth Health 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 clinical predictors cohort experience high risk improved in vivo inhibitor/antagonist insight men mesotrypsin molecular subtypes mouse model novel novel therapeutics outcome forecast polypeptide potential biomarker prognostic prognostic assays prognostic tool promoter prostate cancer cell protein expression prototype research study targeted treatment therapeutic target three dimensional cell culture tissue biomarkers tool treatment strategy tumor growth tumor progression

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）：我们已经鉴定出丝氨酸蛋白酶 PRSS3/mesotrypsin 作为一种分子，可预测前列腺切除术后前列腺癌的全身进展，并且在转移性前列腺癌组织中表达上调。我们还发现，在原位小鼠模型中，前列腺癌细胞中胰岛蛋白酶表达的沉默会抑制培养物中的侵袭性和转移，而用重组胰岛蛋白酶处理前列腺癌细胞会刺激侵袭行为。我们的初步数据表明，胰间蛋白酶通过裂解细胞外基质 (ECM) 中的一种或多种特定底物，导致 COX-2 上调并刺激侵袭行为，从而促进前列腺癌进展。我们假设胰间蛋白酶可能代表转移性前列腺癌的潜在治疗靶点和系统进展的有用的预后组织生物标志物。在这里，我们提出实验：(1) 进一步明确胰蛋白酶促进前列腺癌进展的生物学机制，(2) 开发有效的选择性胰蛋白酶抑制剂，以促进培养和动物模型的机制研究，并为胰蛋白酶靶向治疗提供候选药物，以及（3）评估胰蛋白酶作为预后组织生物标志物。在目标 1 中，我们将使用蛋白质组学方法鉴定 ECM 中胰蛋白酶的蛋白水解底物，并使用 3D 培养模型测试它们对侵袭的影响。我们还将使用启动子报告基因检测和 RNA 干扰方法确定胰蛋白酶调节 COX-2 转录的机制。在目标 2 中，我们将采用结构引导的蛋白质工程来优化胰蛋白酶的多肽抑制剂，并评估该抑制剂对培养测定中的侵袭以及原位小鼠模型中的肿瘤生长和转移的影响。在目标 3 中，我们将使用新开发的选择性胰蛋白酶抗体，评估胰蛋白酶蛋白染色作为高风险根治性前列腺切除术队列中潜在的预后组织生物标志物。我们还将使用不同的转移组织来确定胰蛋白酶表达是否与特定器官部位的转移相关。这些目标的成功完成将极大地提高我们对前列腺癌进展的新分子机制的理解。