Serine Protease Modulation of ErbB Receptors

ErbB 受体的丝氨酸蛋白酶调节

• 批准号: 8099942
• 负责人: KARL X CHAI
• 金额: $ 31.14万
• 依托单位: UNIVERSITY OF CENTRAL FLORIDA
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8099942
• 关键词: Address; Aggressive behavior; American; Anchorage-Independent Growth; Breast Cancer Cell; Cancer Patient; Cancer cell line; Cell Line; Cells; Cleaved cell; Clinical; Combined Modality Therapy; Complementary DNA; Data; Development; Diagnosis; Distant Metastasis; Drug Delivery Systems; Drug resistance; Effectiveness; Engineering; Environment; Enzymes; Evaluation; Extracellular Domain; Functional disorder; Goals; Her2/erbb2/neu Staining Method; Human; In Vitro; Intervention; Investigation; Laboratories; Malignant Neoplasms; Maps; Mediating; Membrane; Modeling; Monoclonal Antibodies; Outcome; Patients; Peptide Hydrolases; Pharmaceutical Preparations; Phenotype; Physiology; Plasmin; Plasminogen Inactivators; Play; Prevention; Protease Inhibitor; Receptor Protein-Tyrosine Kinases; Research; Resistance; Roche brand of trastuzumab; Role; Serine Protease; Signal Transduction; Site; Subfamily lentivirinae; Testing; Transfection; Trastuzumab; Tyrosine Kinase Inhibitor; Up-Regulation; Urokinase; Urokinase Plasminogen Activator Receptor; Woman; chemotherapy; clinically relevant; effective therapy; established cell line; extracellular; hepsin; human PRSS8 protein; inhibitor/antagonist; lapatinib; malignant breast neoplasm; matriptase; meetings; migration; novel; prevent; receptor; research study; resistance mechanism; small molecule; success; treatment strategy; tumor; vector

DESCRIPTION (provided by applicant): The Her2+/uPA+ breast cancers are clinically the most aggressive, more so than the Her2+/uPA-, Her2-/uPA+, and Her2-/uPA- subtypes. Proteolytic modulation of Her2/neu/ErbB2 receptor tyrosine kinase signaling via extracellular domain/ectodomain (ECD) shedding by the urokinase-type plasminogen activator (uPA) and a novel network of extracellular membrane serine proteases is hypothesized to play a mechanistic role in the aggressive behaviors of the Her2+/uPA+ breast cancers and in resistance to anti-Her2 therapies using monoclonal antibody or small-molecule tyrosine kinase inhibitor drugs. Targeting Her2 and uPA simultaneously in Her2+/uPA+ breast cancers could therefore be a more effective strategy of treating these cancers; and potentially all Her2+ breast cancers. This hypothesis will be tested in this project. With the following specific aims, we will be creating novel cell-line models of Her2+/uPA+ breast cancer and testing the effectiveness of a novel treatment strategy by targeting Her2 and uPA simultaneously. We will also be investigating novel functional roles of several extracellular membrane serine proteases associated with breast cancer, and many other cancers. Specific Aim 1. To establish Her2+/uPA+ human breast cancer cell lines for evaluation of enhanced aggressiveness. We will establish Her2+/uPA+ human breast cancer cell lines by over-expressing uPA/uPAR in the Her2-amplified SK-BR-3 and BT-474 cell lines or over-expressing Her2 in the uPA+ but Her2-negative (unamplified) MDA-MB-231 cell line; and perform in vitro evaluations of tumor aggressiveness in proliferation, migration, invasion, and anchorage-independent growth. Specific Aim 2. To treat Her2+/uPA+ human breast cancer cell lines with Herceptin, lapatinib, and WX-UK1 for evaluation of effectiveness of targeting Her2 and uPA together. With our engineered Her2+/uPA+ model cell lines, the following clinically relevant questions will be addressed: 1). whether Her2+/uPA+ breast cancers present de novo drug resistance to Herceptin or even lapatinib and whether uPA inhibition with a selective uPA inhibitor WX-UK1 overcomes the resistance; 2) whether Her2+/uPA+ breast cancers acquire resistance to anti- Her2 therapies more aggressively and whether uPA inhibition with WX-UK1 delays the acquisition of resistance. Specific Aim 3. To determine the specific actions of matriptase, prostasin, and uPA on Her2 ECD. We will use the FT-293 cell line in co-transfection experiments to determine if uPA is a Her2 ECD sheddase and if plasmin plays a role in Her2 ECD shedding; to purify protease-cleaved Her2 for mapping the cleavage sites; and to determine if the protease-cleaved Her2 is responsive to Herceptin-induced turnover. We will use the SK-BR-3 cell line to determine if the Her2 ECD shedding associated with uPA induction in breast cancer cells also involves or requires matriptase and prostasin by using specific inhibitors. PUBLIC HEALTH RELEVANCE: Resistance to chemotherapy drugs targeting Her2/neu/ErbB2 limits the success of these drugs on select groups of breast cancer patients and could render patients with acquired resistance untreatable. Proteolytic shedding of the Her2 extracellular domain/ectodomain (ECD) by proteases is a mechanism of this resistance. Our preliminary studies have identified new candidate proteases involved in Her2 ECD shedding and the proposed research will reveal if specific protease inhibitors should be considered for combination therapy to reduce or prevent the resistance to the anti-Her2 drugs.

描述（由申请人提供）：HER2+/UPA+乳腺癌在临床上是最具侵略性的，比HER2+/UPA-，HER2-/UPA+和HER2-/UPA-亚型更重要。通过细胞外结构域/胞外域（ECD）通过尿激酶型纤溶酶原激活剂（UPA）和细胞外膜膜蛋白酶的新型网络，通过细胞外结构域/ECD域（ECD）脱落的HER2/NEU/ERBB2受体酪氨酸激酶信号传导的蛋白质水解调节。 HER2+/UPA+乳腺癌的行为以及使用单克隆抗体或小分子酪氨酸激酶抑制剂药物的抗HER2疗法的抗性。因此，在HER2+/UPA+乳腺癌中同时靶向HER2和UPA可能是治疗这些癌症的更有效策略。并可能是所有HER2+乳腺癌。该假设将在该项目中进行检验。以以下特定目标，我们将创建HER2+/UPA+乳腺癌的新型细胞系模型，并通过同时靶向HER2和UPA来测试新型治疗策略的有效性。我们还将研究与乳腺癌和许多其他癌症相关的几种细胞外膜丝氨酸蛋白酶的新功能作用。具体目标1。建立HER2+/UPA+人类乳腺癌细胞系以评估增强的攻击性。我们将通过在HER2放大的SK-BR-3和BT-474细胞系中过度表达UPA/UPA来建立HER2+/UPA+人类乳腺癌细胞系或在UPA+中过表达HER2，但HER2-DEGATER（未扩增）MDA MDA -MB-231细胞系；并在体外评估肿瘤在增殖，迁移，侵袭和无关生长中的肿瘤侵袭性。特定目的2。用赫赛汀，拉帕替尼和WX-IK1处理HER2+/UPA+人类乳腺癌细胞系，以评估靶向HER2和UPA的有效性。借助我们设计的HER2+/UPA+模型细胞系，将解决以下临床相关问题：1）。 HER2+/UPA+乳腺癌是否表现出对Herceptin甚至Lapatinib的抗药性，以及使用有选择性UPA抑制剂WX-IK1抑制UPA是否克服了抵抗力； 2）HER2+/UPA+乳腺癌是否更积极地获得对抗HER2疗法的抵抗力，以及使用WX-IK1抑制UPA是否会延迟获得抵抗力。具体目的3。确定Matriptase，Prostasin和UPA对HER2 ECD的特定作用。我们将在共转染实验中使用FT-293细胞系来确定UPA是否为HER2 ECD SHEDDASE，以及纤溶酶是否在HER2 ECD脱落中起作用；净化蛋白酶切割的HER2以绘制裂解位点；并确定蛋白酶裂开的HER2是否对赫赛汀引起的营业额有反应。我们将使用SK-BR-3细胞系来确定与乳腺癌细胞中与UPA诱导相关的HER2 ECD脱落还涉及或需要使用特定的抑制剂，或者需要matriptase和Prostasin。 公共卫生相关性：针对HER2/NEU/ERBB2的化学疗法药物的抗性将这些药物的成功限制在某些乳腺癌患者中，并可能使获得性耐药性患者无法治疗。蛋白酶的HER2细胞外结构域/胞外域（ECD）的蛋白水解脱落是这种抗性的机制。我们的初步研究已经确定了与HER2 ECD脱落有关的新候选蛋白酶，拟议的研究将揭示是否应考虑使用特定的蛋白酶抑制剂进行联合疗法，以降低或防止抗抗HER2药物的耐药性。

项目成果

Ibuprofen regulates the expression and function of membrane-associated serine proteases prostasin and matriptase.

• DOI: 10.1186/s12885-015-2039-6
• 发表时间: 2015-12-29
• 期刊: BMC cancer
• 影响因子: 3.8
• 作者: Chai AC;Robinson AL;Chai KX;Chen LM
• 通讯作者: Chen LM