Combination therapy to inhibit glioma growth by Ad MMP2 and radiation

描述（由申请人提供）：恶性脑肿瘤是最难治疗的癌症之一，并且仍然无法治愈。神经胶质瘤是最常见的脑肿瘤类型，并且以各种等级发生，患者的预后与等级成反比。放射治疗仍然是控制恶性和良性脑膜瘤治疗方式的主要组成部分。在残留或复发的良性肿瘤患者中，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，如放射外科手术，也可能发生放射相关的副作用。尽管有一定的治疗效果，最近的证据表明，辐射可能会促进肿瘤细胞的恶性行为在体外和体内激活参与肿瘤的侵袭，血管生成和转移的几个途径。包括我们在内的几项研究表明，MMP-2在恶性胶质瘤或胶质母细胞瘤中的水平显著升高。我们希望在靶向MMP-2基因的同时，联合放射治疗，观察其抗肿瘤作用。该提案的具体目标是：具体目标1。在体外和体内模型中评估p-MMP-2构建体和辐射单独或组合对胶质瘤细胞生长、侵袭和血管生成的影响。目标1a。确定p-MMP-2和照射单独或联合对胶质母细胞瘤细胞系中MMP-2水平的影响。目标1b。确定p-MMP-2和辐射单独或联合对体外模型中人胶质母细胞瘤细胞系侵袭行为的影响，以及对照/模拟和SV（乱序载体）的影响。目标1c。评价p-MMP-2和照射单独或联合对体外和体内脑血管生成的影响。目标1d。确定p-MMP-2和辐射单独或联合对预先建立的颅内肿瘤生长或裸鼠脑内注射的人胶质母细胞瘤细胞系侵袭性的最佳剂量。具体目标2。确定p-MMP-2和辐射单独或联合对胶质母细胞瘤细胞系增殖、迁移、粘附和凋亡的分子机制的影响。目标2a。与对照/模拟和p-SV对照相比，研究p-MMP-2和辐射单独或联合对胶质母细胞瘤细胞系中粘附和迁移的分子机制的影响。目标2b。与模拟和p-SV相比，确定p-MMP-2和辐射单独或组合对胶质母细胞瘤细胞系增殖的分子机制的影响。目标2c。与模拟和p-SV相比，评价p-MMP-2和辐射单独或联合对胶质母细胞瘤细胞系凋亡分子机制的影响。拟议中的研究应产生重大的见解，放射诱导的改变后，MMP-2的抑制肿瘤的发病机制，反过来，应建议新的治疗干预胶质母细胞瘤的目标。公共卫生相关性：尽管采取了许多治疗策略来治疗多形性胶质母细胞瘤，但患有这种侵袭性脑恶性肿瘤的患者的存活率仍然很低。25年来，受累野放疗一直是胶质母细胞瘤最有效的治疗方法。然而，越来越多的长期生存者与继发性副作用，从这种治疗强调需要开发新的治疗方法。该提议代表了使用p-MMP-2 siRNA的组合治疗方法。这种策略可能会改善胶质母细胞瘤的放射治疗结果。

DESCRIPTION (provided by applicant): Malignant brain tumors represent one of the most refractory cancers to therapy and remain incurable. Gliomas represent the most common type of brain tumors and occur in various grades, with the patient's prognosis inversely proportional to grade. Radiotherapy remains a major component of treatment modalities for controlling both malignant and benign meningiomas. 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. Several studies, including ours, demonstrated significantly increased levels of MMP-2 in malignant gliomas or glioblastomas. We hopothesize that the MMP-2 gene will be targeted while simultaneously combined with radiation and their anti-cancer effects will be determined. The specific aims of this proposal are: Specific Aim 1. Evaluate the effect of p-MMP-2 construct and irradiation alone, or in combination, on glioma cell growth, invasion and angiogenesis in both in vitro and in vivo models. Aim 1a. Determine the effect of p-MMP-2 and irradiation alone, or in combination, on the levels of MMP-2 in glioblastoma cell lines. Aim 1b. Determine the effect of p-MMP-2 and irradiation alone, or in combination, on the invasive behavior of human glioblastoma cell lines in in vitro models with that of controls/mock and SV (scrambled vector). Aim 1c. Evaluate the effect of p-MMP-2 and irradiation alone, or in combination, on cerebral angiogenesis both in vitro and in vivo. Aim 1d. Determine the optimal doses of p-MMP-2 and irradiation alone, or in combination, on pre-established intracranial tumor growth or invasiveness of human glioblastoma cell lines injected intracerebrally in nude mice. Specific Aim 2. Determine the effect of p-MMP-2 and irradiation alone, or in combination, on the molecular mechanisms of proliferation, migration, adhesion and apoptosis in glioblastoma cell lines. Aim 2a. Investigate the effect of p-MMP-2 and irradiation alone, or in combination, on the molecular mechanisms of adhesion and migration in glioblastoma cell lines compared to control/mock and p-SV controls. Aim 2b. Determine the effect of p-MMP-2 and irradiation alone, or in combination, on the molecular mechanisms of proliferation in glioblastoma cell lines compared with mock and p-SV. Aim 2c. Evaluate the effect of p-MMP-2 and irradiation alone, or in combination, on the molecular mechanisms of apoptosis in glioblastoma cell lines compared with mock and p-SV. The proposed studies should generate major insights into the pathogenesis of radiation-induced alteration in tumor after inhibition of MMP-2 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. Involved-field 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 from this treatment highlighted the need for development of novel therapeutic approaches. This proposal represents a combinational therapeutic approach using p-MMP-2 siRNA. This strategy may improve radiotherapy outcomes for the treatment of glioblastomas.