Interaction between Tissue Factor, Junctional Adhesion Molecule-A, and Integrin B1 to drive self-renewal in glioblastoma

组织因子、连接粘附分子 A 和整合素 B1 之间的相互作用驱动胶质母细胞瘤的自我更新

• 批准号: 10554404
• 负责人: Craig Michael Horbinski
• 金额: $ 19.79万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10554404
• 关键词: AKT1 gene; Adhesions; Adult; Aftercare; Back; Behavior; Binding; Blood coagulation; Brain; Caring; Cell Adhesion Molecules; Cell Maintenance; Cell-Cell Adhesion; Cells; Chemotherapy and/or radiation; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Coagulation Process; Complex; Cytotoxic Chemotherapy; Data; Diagnosis; Disease; Epidermal Growth Factor Receptor; Excision; FOS gene; Foundations; G-Protein-Coupled Receptors; Gene Silencing; Genes; Genetic Transcription; Glioblastoma; Growth; Hemorrhage; Hemostatic function; Immunoprecipitation; Impairment; In Vitro; Integrin Inhibition; Integrins; JUN gene; Knock-out; Laboratories; MAP Kinase Gene; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Membrane; Molecular; Normal tissue morphology; Oncogenic; Operative Surgical Procedures; Outcome; PAR-2 Receptor; PIK3CG gene; Pathway interactions; Patient-Focused Outcomes; Patients; Persons; Platelet-Derived Growth Factor Receptor; Platelet-Derived Growth Factor beta Receptor; Protein Secretion; Proteins; Publishing; Receptor Activation; Receptor Protein-Tyrosine Kinases; Recurrent tumor; Resistance; Role; Serine Proteinase Inhibitors; Serpins; Signal Transduction; Testing; Therapeutic; Thromboplastin; Tissue Model; Toxic effect; Translations; United States; Work; c-myc Genes; cancer recurrence; cancer stem cell; cell behavior; chromatin immunoprecipitation; conventional therapy; genotoxicity; improved; in vivo; junctional adhesion molecule; knock-down; neoplastic cell; new therapeutic target; novel therapeutic intervention; overexpression; pharmacologic; programs; promoter; response; self-renewal; stem cell population; stem cell self renewal; stem cell survival; stemness; success; therapeutic target; therapeutically effective; transcription factor; tumor; tumor eradication

PROJECT SUMMARY Glioblastoma (GBM) is the most common cancer arising in the adult brain, and is lethal in nearly all cases. A key contributing factor to poor outcomes for GBM patients is a subpopulation of cells, known as cancer stem cells (CSCs), that are highly resistant to routinely used genotoxic/cytotoxic therapies, and ultimately manifest as recurrent tumor. Inhibiting tumor recurrence from CSCs that survive therapy would therefore improve GBM patient outcomes. Our preliminary and recently published work show that CSC survival is dependent on Tissue Factor (TF), a conserved transmembrane and secreted protein involved in blood clotting. TF activates protease- activated receptor 2 (PAR2), a G-protein-coupled receptor on GBM cells, which promotes CSC maintenance and expansion, as indicated by analysis of marker expression, self-renewal capacity, and in vivo growth of CSCs. TF suppression greatly reduces CSC subpopulations, in some cases even leading to complete tumor eradication in vivo. Protective and proliferative effects of TF-PAR2 signaling on CSCs appears to be through activation of multiple classes of oncogenic receptor tyrosine kinases (RTKs) like EGFR, although the mechanism by which TF-PAR2-RTKs stimulate CSCs is unclear. Our preliminary data also show that TF positively correlates with expression of Junctional Adhesion Molecule-A (JAM-A), a protein that promotes cell-cell adhesion by stabilizing integrin β1. CSCs depend on such cell-cell adhesion, and our data show that JAM-A is necessary for CSC behavior in GBM. Because TF can also signal through integrin β1, our overarching hypothesis is that TF upregulates JAM-A expression, which stabilizes integrin β1 and enhances the ability of TF to act on integrin β1 and promote self-renewal in GBM. This has therapeutic relevance, because the pro-CSC effects of TF are independent of its role in hemostasis, and blocking JAM-A could potentially reduce the pro-tumor effects of TF, without causing bleeding that would result from targeting TF directly. In Specific Aim 1, we will test the hypothesis that TF drives JAM-A expression through PAR2-RTK signaling. We will identify components of the TF-PAR2 complex that are essential for TF-PAR2 pro-tumor activities, and will test the ability of TF to trigger JAM-A expression while inhibiting each major downstream pathway of RTK signaling. In Specific Aim 2, we will determine whether JAM-A requires serpin B3, a serine-protease inhibitor that we have shown binds to JAM-A, for its pro-CSC activities. In Specific Aim 3, we will use molecular knockouts to determine whether JAM-A is an effective therapeutic target against aggressive, high TF-expressing GBM. Completion of this project will advance our understanding of how CSC subpopulations are maintained in GBM, could well be applicable to a wide range of cancers, and would demonstrate a compelling new therapeutic target in treating numerous malignancies, including GBM.

项目概要 胶质母细胞瘤（GBM）是成人大脑中最常见的癌症，几乎在所有病例中都是致命的。 导致 GBM 患者预后不良的因素是细胞亚群，称为癌症干细胞 （CSC），对常规使用的基因毒性/细胞毒性疗法具有高度抵抗力，最终表现为 因此，抑制治疗后存活的 CSC 的肿瘤复发将改善 GBM。 我们的初步和最近发表的工作表明，CSC 的存活取决于组织。 因子 (TF) 是一种参与血液凝固的保守跨膜和分泌蛋白。 激活受体 2 (PAR2)，GBM 细胞上的一种 G 蛋白偶联受体，可促进 CSC 维持 通过对 CSC 的标记表达、自我更新能力和体内生长的分析表明。 TF 抑制大大减少了 CSC 亚群，在某些情况下甚至导致肿瘤完全根除 TF-PAR2 信号传导对 CSC 的保护和增殖作用似乎是通过激活 多种致癌受体酪氨酸激酶 (RTK)，如 EGFR，尽管其机制 TF-PAR2-RTK 刺激 CSC 的情况尚不清楚。我们的初步数据还表明 TF 与 CSC 呈正相关。 连接粘附分子-A (JAM-A) 的表达，这是一种通过稳定细胞间粘附来促进细胞间粘附的蛋白质 CSC 依赖于这种细胞间粘附，我们的数据表明 JAM-A 对于 CSC 是必需的。 由于 TF 也可以通过整合素 β1 发出信号，因此我们的首要假设是 TF 上调 JAM-A 表达，从而稳定整合素 β1 并增强 TF 作用于整合素 β1 的能力 并促进 GBM 的自我更新，这具有治疗相关性，因为 TF 的促 CSC 作用是 与其在止血中的作用无关，并且阻断 JAM-A 可能会降低 TF 的促肿瘤作用， 不会造成直接针对 TF 导致的出血。在具体目标 1 中，我们将检验该假设。 TF 通过 PAR2-RTK 信号传导驱动 JAM-A 表达 我们将鉴定 TF-PAR2 的组成部分。 复合物对于 TF-PAR2 促肿瘤活性至关重要，并将测试 TF 触发 JAM-A 的能力 在具体目标 2 中，我们将抑制 RTK 信号传导的每个主要下游途径。 确定 JAM-A 是否需要丝氨酸蛋白酶抑制剂 B3，我们已证明它是一种丝氨酸蛋白酶抑制剂，可与 JAM-A 结合， 在特定目标 3 中，我们将使用分子敲除来确定 JAM-A 是否是一种促 CSC 活性。 针对侵袭性、高 TF 表达的 GBM 的有效治疗靶点将推进该项目的完成。 我们对 GBM 中 CSC 亚群如何维持的理解很可能适用于广泛的范围 癌症，并将在治疗多种恶性肿瘤中展示出令人信服的新治疗靶点， 包括GBM。