Cath B and uPAR si RNA constructs regressed glioma growth

Cath B 和 uPAR si RNA 构建使神经胶质瘤生长退化

• 批准号: 7982701
• 负责人: JASTI S. RAO
• 金额: $ 15.14万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-06-01 至 2015-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7982701
• 关键词: Adhesions; Adverse effects; Antisense Oligonucleotides; Apoptosis; Apoptotic; Basement membrane; Behavior; Benign; Biologic Characteristic; Brain Neoplasms; Cathepsins B; Cell Adhesion; Cell Death; Cell Line; Cell Proliferation; Cells; Cerebrum; Characteristics; Clinical Trials; DNA Repair; DNA damage checkpoint; Data; Development; Dose; Down-Regulation; Embryo; Enzymes; Extracellular Matrix Degradation; Genes; Glioblastoma; Glioma; Growth; Human; Immigration; Immune response; In Vitro; Intracranial Neoplasms; Invaded; Laboratories; Maintenance; Malignant - descriptor; Malignant Glioma; Malignant Neoplasms; Mediating; Membrane Proteins; Messenger RNA; Modality; Molecular; Morbidity - disease rate; Neoplasm Metastasis; Nude Mice; Oligonucleotides; Pathogenesis; Pathway interactions; Patients; Peptide Hydrolases; Population; Primary Brain Neoplasms; Process; RNA; RNA Interference; Radiation; Radiation therapy; Radiosurgery; Receptor Gene; Receptor Inhibition; Recurrence; Regulation; Research; Research Personnel; Residual state; Resistance; Role; Signal Transduction; Small Interfering RNA; Specificity; Stem cells; Survival Rate; Survivors; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Tissues; Treatment outcome; Tumor Cell Invasion; Urokinase Plasminogen Activator Receptor; Xenograft procedure; angiogenesis; base; brain tissue; cancer cell; cancer stem cell; cancer therapy; cell growth; cytotoxic; effective therapy; glioma cell line; improved; in vitro Model; in vivo; inhibitor/antagonist; insight; irradiation; malignant phenotype; migration; mortality; neoplastic cell; nerve stem cell; novel; novel therapeutic intervention; outcome forecast; prevent; public health relevance; receptor downregulation; receptor expression; self-renewal; treatment effect; tumor; tumor growth; tumor progression; tumorigenic; vector

DESCRIPTION (provided by applicant): Despite the many therapeutic strategies undertaken for treatment of glioblastoma multiforme, the survival rate remains low for patients afflicted with this aggressive cerebral malignancy. Even with the combined use of several therapeutic modalities, a good prognosis is extremely rare as the remaining cancer cells inevitably infiltrate the normal brain tissue and cause tumor recurrence. Local invasion remains an important cause of morbidity and mortality from brain tumors. The long-term objective of my laboratory is to understand the cellular and molecular mechanisms that underlie tumor invasiveness in human gliomas. Radiotherapy remains a major component of treatment modalities for controlling both malignant and benign glioblastomas. In patients with residual or recurring benign tumors, there is increasing concern about radiation-related side effects that may occur even with highly accurate therapies such as radiosurgery. Despite some therapeutic effect, recent evidence has shown that irradiation may promote malignant behaviors of cancer cells both in vitro and in vivo by activating several pathways involved in tumor invasiveness, angiogenesis and metastasis. Further, the glioma initiating cells are resistant to radiotherapy. Several studies, including ours, demonstrated that significantly increased levels of cath B and uPAR in malignant gliomas and radiation further activates cath B and uPAR molecules. We hypothesize that the cath B and uPAR gene will be targeted while simultaneously combined with radiation and their anti-cancer effects will be determined. The specific aims of the proposal are: Specific Aim 1. Evaluate the effect of vectors expressing siRNA targeting cathepsin B and uPAR (pCU) on glioma cell adhesion, migration and invasion in in vitro models with and without radiation. Aim 1a. Determine the effect of pC, pU, pCU and radiation alone and in combination on the levels of cath B and uPAR in glioblastoma cell lines and glioma initiating cells (GIC). Aim 1b. Determine the downregulation of cath B and uPAR on glioma cell adhesion and migration in glioblastoma cell lines and glioma initiating cells with and without radiation. Aim 1c. Evaluate the effect of pC, pU, pCU and radiation alone and in combination on the invasive behavior of human glioblastoma cell lines and GIC. Specific Aim 2. Determine the molecular mechanisms in cath B and uPAR inhibition-mediated changes on cell growth, proliferation and apoptosis in GIC and non-GIC with and without radiation. Aim 2a. Evaluate the molecular mechanisms in cath B and uPAR downregulation on cell growth and proliferation with and without radiation in non-GIC and GIC. Aim 2b. Determine the effect of cath B and uPAR downregulation on the molecular mechanisms of apoptosis in glioblastoma cell lines and GIC with and without radiation. Specific Aim 3. Determine the molecular mechanisms in cath B and uPAR inhibition-mediated changes in angiogenesis and tumor growth in non-GIC and GIC with and without radiation in nude mice. Aim 3a. Evaluate the effect of cath B and uPAR downregulation-mediated changes in the signaling mechanisms in the regulation of cerebral angiogenesis both in vitro and in vivo with and without radiation. Aim 3b. Determine the optimal doses of pC, pU, pCU and radiation alone and in combination on pre-established intracranial tumor growth or the invasiveness of human glioblastoma cell lines and GIC injected intracerebrally in nude mice. The proposed studies should generate major insight into the pathogenesis of cath B and uPAR and, in turn, should suggest novel targets for therapeutic interventions of glioblastomas. PUBLIC HEALTH RELEVANCE: Despite the many therapeutic strategies undertaken for treatment of glioblastoma multiforme, the survival rate for patients afflicted with this aggressive cerebral malignancy remains low. Radiotherapy has remained the single most effective therapy of glioblastoma for more than 25 years. However, an increasing number of long survivors with secondary side effects and the resistance of glioma initiating cells from this treatment highlighted the need for development of novel therapeutic approaches. This proposal represents a combinational therapeutic approach using a single and bicistronic siRNA construct for cathepsin B and uPAR. This strategy may improve radiotherapy outcomes for the treatment of glioblastomas.

描述（由申请人提供）：尽管有许多治疗策略用于治疗多形性胶质母细胞瘤，但这种侵袭性脑恶性肿瘤患者的生存率仍然很低。即使联合使用几种治疗方式，由于剩余的癌细胞不可避免地渗入正常脑组织并引起肿瘤复发，预后良好的情况极为罕见。局部侵袭仍然是脑肿瘤发病和死亡的重要原因。我实验室的长期目标是了解人类胶质瘤侵袭性的细胞和分子机制。放疗仍然是控制恶性和良性胶质母细胞瘤治疗方式的主要组成部分。对于残留或复发的良性肿瘤患者，人们越来越关注放射相关的副作用，即使使用高度精确的治疗方法，如放射外科手术，也可能发生。尽管有一定的治疗效果，但最近的证据表明，辐照可能通过激活涉及肿瘤侵袭、血管生成和转移的几种途径，在体外和体内促进癌细胞的恶性行为。此外，胶质瘤起始细胞对放射治疗有抵抗性。包括我们在内的几项研究表明，恶性胶质瘤中cath B和uPAR水平显著升高，辐射进一步激活了cath B和uPAR分子。我们假设cath B和uPAR基因将在与放疗同时结合的情况下被靶向，并确定它们的抗癌作用。该提案的具体目标是：在有和无辐射的体外模型中，评估表达siRNA靶向组织蛋白酶B和uPAR （pCU）的载体对胶质瘤细胞粘附、迁移和侵袭的影响。目的1。测定pC、pU、pCU和辐射单独或联合对胶质母细胞瘤细胞系和胶质瘤起始细胞（GIC）中cath B和uPAR水平的影响。目标1 b。测定放射线和非放射线对胶质瘤细胞系和胶质瘤起始细胞中cath B和uPAR对胶质瘤细胞粘附和迁移的下调作用。目标1 c。评价pC、pU、pCU和辐射单独或联合作用对人胶质母细胞瘤细胞系和GIC侵袭行为的影响。具体目标2。确定cath B和uPAR抑制介导的GIC和非GIC细胞生长、增殖和凋亡变化的分子机制。目标2。评估cath B和uPAR下调对非GIC和GIC细胞生长和增殖的分子机制。目标2 b。探讨cath B和uPAR下调对胶质母细胞瘤细胞系和GIC细胞凋亡分子机制的影响。具体目标3。确定cath B和uPAR抑制介导的非GIC和GIC裸鼠血管生成和肿瘤生长变化的分子机制。目标3。评估cath B和uPAR下调介导的信号机制变化在体外和体内有和无辐射情况下对脑血管新生的调节作用。目标3 b。确定pC、pU、pCU和辐射单独或联合使用对裸鼠颅内肿瘤生长或人胶质母细胞瘤细胞系侵袭性的最佳剂量和脑内注射GIC。拟议的研究应该对cath B和uPAR的发病机制产生重要的见解，反过来，应该为胶质母细胞瘤的治疗干预提供新的靶点。