Inhibition of matrix proteases to sensitize medulloblastoma cells to radiation

抑制基质蛋白酶使髓母细胞瘤细胞对辐射敏感

• 批准号: 8211080
• 负责人: JASTI S. RAO
• 金额: $ 31.6万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8211080
• 关键词: Address; Adhesions; Adult; Apoptosis; Area; Astrocytoma; Attention; Back; Basic Science; Behavior; Brain; Brain Neoplasms; Cells; Cerebellum; Cerebrum; Child; Clinical; Development; Disease; Dose; Enzymes; Excision; External Beam Radiation Therapy; Gelatinase A; Gelatinase B; Genetic Materials; Glioma; Goals; Growth; Human; Immigration; Immune response; In Vitro; Infant; Infiltration; Intracranial Neoplasms; Invaded; Lead; Learning; Malignant - descriptor; Malignant Neoplasms; Matrix Metalloproteinases; Memory; Messenger RNA; Metalloproteases; Molecular; Morbidity - disease rate; Neoplasm Metastasis; Neuraxis; Neuronal Differentiation; Nude Mice; Operative Surgical Procedures; Patients; Peptide Hydrolases; Plasmid Cloning Vector; Plasminogen Activator; Primitive Neuroectodermal Tumor; RNA Interference; Radiation; Radiation therapy; Recurrence; Relapse; Resistance; Small Interfering RNA; Staging; Technology; Therapeutic; Therapeutic Effect; Toxic effect; Urokinase; Urokinase Plasminogen Activator Receptor; Vertebral column; angiogenesis; base; cancer cell; cell growth; cellular targeting; chemotherapy; disability; emotional adjustment; gene therapy; improved; in vitro Model; in vivo; in vivo Model; irradiation; killings; medulloblastoma; medulloblastoma cell line; migration; neoplastic cell; neurosurgery; novel therapeutic intervention; outcome forecast; prevent; research study; response; social; treatment strategy; tumor; tumor growth; tumor progression; vector

DESCRIPTION (provided by applicant): Medulloblastomas, which belong to a group of primitive neuroectodermal tumors, are invasive tumors with predominant neuronal differentiation. Despite technological advances in neurosurgery, chemotherapy and radiation therapy, the prognosis for patients with these tumors remains variable and is relatively poor in infants and adult patients with metastatic disease. The traditional treatments are also toxic and can lead to long-term disabilities. Therefore new strategies are needed to prevent treatment related morbidity in these patients. One avenue possibly worth exploring further is the use of inhibition of matrix enzymes that contribute to collateral damage after therapeutic irradiation. This approach may also prevent clinical progression of the tumor by thwarting the invasive infiltration that characterizes glioma growth. A significant association of urokinase plasminogen activator (uPA) and matrix metalloprotease-9 (MMP-9) expression with survival and M-stage indicates that these proteases may modulate the survival of medulloblastoma patients. The levels of uPAR and MMP-9 expression and cellular invasiveness were increased in irradiated medulloblastoma cells. These findings led us to hypothesize inhibition of uPAR and MMP-9 by RNAi technology could be a potential therapeutic approach to improve the efficacy of radiotherapy in medulloblastoma patients. Specific Aim 1. Evaluate the effect of uPAR and MMP-9 inhibition and radiation alone and in combination on medulloblastoma cell growth, invasion and angiogenesis in both in vitro and in vivo models. Aim 1a. Determine the effect of puPAR, pMMP-9 and pUM in combination with radiation on the levels of uPAR and MMP-9 in medulloblastoma cell lines. Aim 1b. Determine the effect of puPAR, pMMP-9, pUM and radiation alone and in combination on the invasive behavior of human medulloblastoma cell lines in vitro models. Aim 1c. Evaluate the effect of puPAR, pMMP-9, pUM and radiation alone and in combination on cerebral angiogenesis in both in vitro and in vivo models. Aim 1d. Determine the optimal doses of puPAR, pMMP-9, pUM and radiation alone and in combination on pre-established intracranial tumor growth or invasiveness of human medulloblastoma cell lines injected intracerebrally in nude mice. Specific Aim 2. Determine the effect of puPAR, pMMP-9, pUM and radiation alone and in combination on the molecular mechanisms of proliferation, migration, adhesion and apoptosis in medulloblastoma cell lines. Aim 2a. Investigate the effect of puPAR, pMMP-9, pUM and radiation alone in combination on the molecular mechanisms of adhesion and migration in medulloblastoma cell lines compared to control/mock and scrambled vector (pSV) controls. Aim 2b. Determine the effect of puPAR, pMMP-9, pUM and radiation alone and in combination on the molecular mechanisms of proliferation in medulloblastoma cell lines compared with mock and pSV controls. Aim 2c. Evaluate the effect of puPAR, pMMP-9, pUM and in combination on the molecular mechanisms of apoptosis in medulloblastoma cell lines compared with mock and Psv controls. This combination of in vitro basic science experiments and translational in vivo studies will provide the basis for development of a new therapeutic approach to medulloblastoma tumors which are resistant to conventional radiotherapy. PUBLIC HELATH RELEVANCE: Medulloblastomas are one type of brain tumors that are found near the midline of the cerebellum. It is a type of brain tumor that occurs in infants and young children. After surgery, external radiation to the entire CNS (craniospinal irradiation, or CSI) is recommended to prevent the tumor from coming back in this area (recurrence, or relapse). Much attention has understandably been paid to the possible long-term complications of radiation therapy to the brain and spine of a growing child. These can include deficits in memory, learning, and social/emotional adjustment, and growth problems. A major goal for these patients is to develop treatment strategies that minimize the dose of radiation to the central nervous system. Gene therapy consists of the introduction of genetic material into diseased cellular targets to bring about therapeutic benefit. The present study explores the usefulness of gene therapy to inhibit protease expression in conjunction with radiation for killing of medulloblastoma cells with lower toxicity.

描述（由申请人提供）：属于一组原始神经外科肿瘤的髓母细胞瘤是具有主要神经元分化的侵入性肿瘤。尽管神经外科，化学疗法和放射疗法的技术进步，但 这些肿瘤的患者仍然可变，并且在婴儿和成年患者的转移性疾病患者中相对较差。传统治疗方法也有毒，可能导致长期残疾。因此，需要采取新的策略来防止这些患者与治疗相关的发病率。一个可能值得进一步探索的大道是使用抑制基质酶，这些基质酶在治疗辐射后会造成附带损害。这种方法还可以通过挫败胶质瘤生长的侵入性浸润来防止肿瘤的临床进展。尿激酶纤溶酶原活化剂（UPA）和基质金属蛋白酶9（MMP-9）表达与存活和M级的显着关联表明，这些蛋白酶可能调节髓母细胞瘤患者的存活率。在辐照的髓母细胞瘤细胞中，UPAR和MMP-9表达和细胞侵袭性的水平升高。这些发现导致我们假设RNAi技术对UPAR和MMP-9抑制可能是一种潜在的治疗方法，可以提高放射疗法对髓母细胞瘤患者的疗效。具体目标1。单独评估UPAR和MMP-9抑制作用和辐射的影响，以及在体外和体内模型中对髓母细胞瘤细胞生长，侵袭和血管生成的结合。目标1a。确定Pupar，PMMP-9和PUM与辐射的影响对髓母细胞瘤细胞系中UPAR和MMP-9水平的影响。目标1B。确定pupar，pmmp-9，pum和辐射的影响，并结合对人髓母细胞瘤细胞系在体外模型中的侵入性行为的影响。 AIM 1C。评估Pupar，PMMP-9，PUM和辐射的影响，以及在体外和体内模型中对脑血管生成的结合。目标1d。确定pupar，pMMP-9，pum和辐射的最佳剂量，并结合在颅内肿瘤的预先建立的颅内肿瘤生长或人髓母细胞瘤细胞系中注入裸鼠内脑内毛内的浸润性。具体目的2。确定pupar，pmmp-9，pum和辐射的影响，以及在髓母细胞瘤细胞系中增殖，迁移，粘附和凋亡的分子机制的结合。目标2a。与对照/模拟和炒载体（PSV）对照组相比，研究Pupar，PMMP-9，PUM和辐射对髓母细胞瘤细胞系的粘附和迁移的分子机制的影响。目标2B。与模拟和PSV对照相比，单独使用Pupar，PMMP-9，PMMP-9，PUM和辐射对髓母细胞瘤细胞系增殖的分子机制的影响。 AIM 2C。与模拟和PSV对照相比，评估Pupar，PMMP-9，PUM和组合对髓母细胞瘤细胞系凋亡的分子机制的影响。体外基础科学实验和体内翻译研究的这种结合将为开发一种新的治疗方法来开发对常规放射疗法具有抗性的髓母细胞瘤肿瘤。 公共螺旋性相关性：髓母细胞瘤是在小脑中线附近发现的一种类型的脑肿瘤。这是一种在婴儿和幼儿中发生的脑肿瘤。手术后，建议对整个CNS（颅骨辐射或CSI）进行外部辐射，以防止肿瘤回到该区域（复发或复发）。可以理解，人们对放射治疗对大脑和脊柱的可能长期并发症引起了很多关注。这些可以包括记忆，学习和社交/情感调整以及成长问题的缺陷。这些患者的主要目标是制定治疗策略，以最大程度地减少对中枢神经系统的辐射剂量。基因治疗包括将遗传物质引入患病的细胞靶标，以带来治疗益处。本研究探讨了基因治疗对抑制蛋白酶表达的有用性，并结合辐射杀死毒性较低的髓母细胞瘤细胞。