Combination therapy to inhibit glioma growth by Ad MMP2 and radiation

描述(申请人提供)：恶性脑瘤是最难治愈的癌症之一，目前仍无法治愈。胶质瘤是最常见的脑肿瘤类型，发生在不同的级别，患者的预后与级别成反比。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，人们越来越担心放射相关的副作用，即使在放射外科等高度精确的治疗下也可能发生这些副作用。尽管有一定的治疗效果，但最近的证据表明，辐射可能通过激活与肿瘤侵袭、血管生成和转移有关的几条途径，在体外和体内促进癌细胞的恶性行为。包括我们在内的几项研究表明，在恶性胶质瘤或胶质母细胞瘤中，基质金属蛋白酶-2的水平显著增加。我们希望将靶向的基质金属蛋白酶-2基因，同时结合放射和它们的抗癌作用将被确定。本研究的具体目的是：1.在体外和体内模型中评价p-MMP2构建和照射单独或联合应用对胶质瘤细胞生长、侵袭和血管生成的影响。目标1a。确定p-MMP2单独或联合应用对胶质母细胞瘤细胞系中MMP2表达水平的影响。目标1b。采用对照/模拟和SV(Sscbled载体)体外模型，观察p-MMP2单独或联合应用对人脑胶质母细胞瘤细胞系侵袭行为的影响。目标1c。评价p-MMP2单独或联合照射对体外和体内脑血管生成的影响。目标1D。确定p-MMP2单独或联合照射对裸鼠脑内注射的人胶质母细胞瘤细胞系预先建立的颅内肿瘤生长或侵袭力的最佳剂量。具体目的2.确定p-MMP2单独或联合应用对胶质母细胞瘤细胞系增殖、迁移、黏附和凋亡的分子机制的影响。目标2a。与对照组和p-SV对照组比较，研究p-MMP2和照射单独或联合作用对胶质母细胞瘤细胞系黏附和迁移的分子机制的影响。目标2b。确定p-MMP2单独或联合照射对胶质母细胞瘤细胞系增殖的分子机制的影响，并与模型和p-SV进行比较。目标2c。与假手术组和p-SV组比较，评价p-MMP2单独或联合应用对胶质母细胞瘤细胞系凋亡分子机制的影响。建议的研究将对抑制基质金属蛋白酶-2后辐射诱导的肿瘤改变的发病机制产生重要的见解，并反过来为胶质母细胞瘤的治疗干预提供新的靶点。公共卫生相关性：尽管对多形性胶质母细胞瘤采取了许多治疗策略，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，受累野放射治疗一直是胶质母细胞瘤最有效的单一治疗方法。然而，越来越多的长期生存者因这种治疗而产生二次副作用，这突显了开发新的治疗方法的必要性。这项建议代表了一种使用p-MMP2 siRNA的联合治疗方法。这一策略可能会改善治疗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.