Regulation of Serine Protease HtrA1 and Chemoresponse

丝氨酸蛋白酶 HtrA1 和化学反应的调节

• 批准号: 8212505
• 负责人: VIJI SHRIDHAR
• 金额: $ 30.41万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-04-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8212505
• 关键词: Anchorage-Independent Growth; Animal Model; Apoptosis; Apoptotic; Cancer cell line; Caspase; Caspase Inhibitor; Cell Death; Cell Line; Cells; Chemotherapy-Oncologic Procedure; Cisplatin; Co-Immunoprecipitations; Data; Deoxycytidine; Development; Diagnosis; Diagnostic; Disease; Dominant-Negative Mutation; Down-Regulation; Drug resistance; Epigenetic Process; Family; Fluorogenic Substrate; Genomics; Growth; Health; Hypermethylation; In Vitro; Lead; Malignant Neoplasms; Malignant neoplasm of ovary; Mediating; Methods; Methylation; Mitochondria; Modification; Molecular; Neoplastic Cell Transformation; Paclitaxel; Pathway interactions; Patients; Pattern; Peptide Hydrolases; Peptide Library; Pharmaceutical Preparations; Phenotype; Play; Primary Neoplasm; Process; Procollagen; Regulation; Resistance; Role; Screening procedure; Serine; Serine Protease; Serine Proteinase Inhibitors; Site; Staining method; Stains; Stress; Substrate Specificity; Testing; Up-Regulation; Western Blotting; Woman; Yeasts; base; bisulfite; cancer cell; cancer therapy; caspase-3; caspase-9; chemotherapy; clinical remission; cytotoxicity; improved; in vivo; insight; member; multicatalytic endopeptidase complex; mutant; novel; novel therapeutic intervention; pro-apoptotic protein; promoter; research study; response; tool; tumor; tumorigenesis; validation studies; vector; yeast two hybrid system

DESCRIPTION (provided by applicant): The majority of women diagnosed with ovarian cancer ultimately succumb to the disease due to the outgrowth of cells resistant to chemotherapy [4]. An improved understanding of cellular mechanisms of chemoresistance is essential in developing new effective anticancer therapy to overcome the problem of chemoresistance. We have shown that HtrA1, a serine protease with previously unknown function, is down-regulated in a majority of ovarian cancer cell lines and primary tumors. Additional studies indicated that HtrA1 may modulate cell death and anchorage independent growth of ovarian cancer cells [2]. Our more recent data indicate that HtrA1 is upregulated by cisplatin and paclitaxel and suggests that it may contribute to sensitivity to chemotherapy. Increased cell death and chemosensitivity following HtrA1 expression depends on the protease activity of HtrA1, suggesting a role of serine protease activity in programmed cell death (PCD). Moreover, 90% of patients (27/30) with tumors expressing high levels of HtrA1 responded to chemotherapy with complete or partial clinical remissions, compared to 62% (8/13) and 65% (11/17) response rates in tumors with low and moderate levels of HtrA1, respectively. Response to chemotherapy was significantly different between low and high (P = 0.0276) or moderate and high (P = 0.0342) staining groups. These findings identify HtrA1 as a novel modulator of cisplatin-induced cytotoxicity and suggest that loss of HtrA1 in ovarian cancer may contribute to chemoresistance [5]. The objective of this application is to determine the mechanisms by which HtrA1 expression is regulated in cancer and define the role of HtrA1 in programmed cell death and chemoresistance. Based on our preliminary studies, we are proposing the following three hypotheses: first, HtrA1 expression is epigenetically regulated in cancer and is transcriptionally upregulated by chemotherapy in cancer cells when its expression is not silenced by epigenetic mechanisms; second, HtrA1 participates in a "serine proteasome"-mediated PCD that cross-talks with caspase-mediated cell death pathways; and third, targeted degradation of specific substrates by HtrA1 following chemotherapy treatment contributes to chemotherapy-induced cytotoxicity. PUBLIC HEALTH RELEVANCE: Emerging evidence suggests that some of the same changes that contribute to neoplastic transformation also contribute to drug resistance. In particular, the same anti-apoptotic changes that contribute to the transformed phenotype by making cells resistant to the stresses of unfavorable growth conditions and loss of important homeostatic processes also appear to make cells more resistant than they would otherwise be to cancer chemotherapy. Based on this view, studies that improve our understanding of the process of tumorigenesis have the potential to also provide new insight into the problem of drug resistance. Our studies have shown that altered expression of the serine protease HtrA1 modulates chemotherapy induced cytotoxicity. At the conclusion of the proposed studies, we hope to elucidate the significance of HtrA1 down-regulation in ovarian cancer and how this reduced HtrA1 expression contributes to the development of chemoresistant ovarian cancer. A better understanding of the role of HtrA1 in a poorly understood serine protease mediated programmed cell death may contribute to discoveries of new therapeutic approaches to overcome drug resistance. Drug resistant ovarian cancer is a lethal disease, and this project will better define novel mechanisms of drug resistance and programmed cell death in ovarian cancer.

描述（由申请人提供）：大多数被诊断患有卵巢癌的女性最终因对化疗产生耐药性的细胞生长而死于该病[4]。更好地了解化学耐药性的细胞机制对于开发新的有效抗癌疗法以克服化学耐药性问题至关重要。我们已经证明，HtrA1（一种以前未知功能的丝氨酸蛋白酶）在大多数卵巢癌细胞系和原发性肿瘤中表达下调。其他研究表明 HtrA1 可能调节卵巢癌细胞的细胞死亡和贴壁独立生长 [2]。我们最新的数据表明，HtrA1 被顺铂和紫杉醇上调，这表明它可能有助于提高对化疗的敏感性。 HtrA1 表达后细胞死亡和化疗敏感性的增加取决于 HtrA1 的蛋白酶活性，表明丝氨酸蛋白酶活性在程序性细胞死亡 (PCD) 中发挥作用。此外，90% (27/30) 表达高水平 HtrA1 的肿瘤患者对化疗有完全或部分临床缓解的反应，而低水平和中度 HtrA1 肿瘤的反应率分别为 62% (8/13) 和 65% (11/17)。低染色组和高染色组（P = 0.0276）或中染色组和高染色组（P = 0.0342）对化疗的反应存在显着差异。这些发现确定 HtrA1 是顺铂诱导的细胞毒性的新型调节剂，并表明卵巢癌中 HtrA1 的缺失可能导致化疗耐药性 [5]。本申请的目的是确定 HtrA1 表达在癌症中的调节机制，并确定 HtrA1 在程序性细胞死亡和化疗耐药中的作用。基于我们的初步研究，我们提出以下三个假设：首先，HtrA1的表达在癌症中受到表观遗传调节，并且当其表达未被表观遗传机制沉默时，癌细胞中的化疗会转录上调HtrA1表达；其次，HtrA1 参与“丝氨酸蛋白酶体”介导的 PCD，与 caspase 介导的细胞死亡途径相互作用；第三，化疗后 HtrA1 对特定底物的靶向降解有助于化疗诱导的细胞毒性。公共卫生相关性：新出现的证据表明，一些导致肿瘤转化的相同变化也会导致耐药性。特别是，通过使细胞对不利生长条件的压力和重要稳态过程的丧失产生抵抗力而导致表型转化的相同抗凋亡变化似乎也使细胞比其他情况下对癌症化疗具有更强的抵抗力。基于这一观点，提高我们对肿瘤发生过程的理解的研究也有可能为耐药性问题提供新的见解。我们的研究表明，丝氨酸蛋白酶 HtrA1 表达的改变可调节化疗诱导的细胞毒性。在拟议研究的结论中，我们希望阐明 HtrA1 下调在卵巢癌中的重要性，以及 HtrA1 表达的减少如何促进化疗耐药性卵巢癌的发展。更好地了解 HtrA1 在人们知之甚少的丝氨酸蛋白酶介导的程序性细胞死亡中的作用可能有助于发现克服耐药性的新治疗方法。耐药性卵巢癌是一种致命的疾病，该项目将更好地定义卵巢癌中耐药性和程序性细胞死亡的新机制。

项目成果

Expression and functional significance of HtrA1 loss in endometrial cancer.

• DOI: 10.1158/1078-0432.ccr-09-3069
• 发表时间: 2011-02-01
• 期刊: Clinical cancer research : an official journal of the American Association for Cancer Research
• 作者: Mullany SA;Moslemi-Kebria M;Rattan R;Khurana A;Clayton A;Ota T;Mariani A;Podratz KC;Chien J;Shridhar V
• 通讯作者: Shridhar V

HtrA serine proteases as potential therapeutic targets in cancer.

HTRA丝氨酸蛋白酶作为癌症的潜在治疗靶标。

• DOI: 10.2174/156800909788486704
• 发表时间: 2009-06
• 期刊: Current cancer drug targets
• 影响因子: 3
• 作者: Chien J;Campioni M;Shridhar V;Baldi A
• 通讯作者: Baldi A

Identification of tubulins as substrates of serine protease HtrA1 by mixture-based oriented peptide library screening.

• DOI: 10.1002/jcb.22121
• 发表时间: 2009-05-15
• 期刊: JOURNAL OF CELLULAR BIOCHEMISTRY
• 影响因子: 4
• 作者: Chien, Jeremy;He, Xiaoping;Shridhar, Viji
• 通讯作者: Shridhar, Viji

Downregulation of HtrA1 promotes resistance to anoikis and peritoneal dissemination of ovarian cancer cells.

• DOI: 10.1158/0008-5472.can-09-3557
• 发表时间: 2010-04-15
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: He X;Ota T;Liu P;Su C;Chien J;Shridhar V
• 通讯作者: Shridhar V

PG545 enhances anti-cancer activity of chemotherapy in ovarian models and increases surrogate biomarkers such as VEGF in preclinical and clinical plasma samples.

• DOI: 10.1016/j.ejca.2015.02.007
• 发表时间: 2015-05
• 期刊: EUROPEAN JOURNAL OF CANCER
• 影响因子: 8.4
• 作者: Winterhoff, Boris;Freyer, Luisa;Hammond, Edward;Giri, Shailendra;Mondal, Susmita;Roy, Debarshi;Teoman, Attila;Mullany, Sally A.;Hoffmann, Robert;von Bismarck, Antonia;Chien, Jeremy;Block, Matthew S.;Millward, Michael;Bampton, Darryn;Dredge, Keith;Shridhar, Viji
• 通讯作者: Shridhar, Viji