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，并且仍然是无法治愈的。胶质瘤是最常见的脑肿瘤类型，可分为不同级别，患者的预后与级别成反比。放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。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。包括我们在内的几项研究表明，恶性胶质瘤或胶质母细胞瘤中MMP-2水平显著升高。我们推测MMP-2基因将被靶向，同时与辐射结合，并确定其抗癌效果。本建议的具体目标是：在体外和体内模型中评估p-MMP-2构建和单独或联合照射对胶质瘤细胞生长、侵袭和血管生成的影响。目的1。确定p-MMP-2和单独或联合照射对胶质母细胞瘤细胞系中MMP-2水平的影响。目标1 b。确定p-MMP-2和辐照单独或联合对体外模型中人类胶质母细胞瘤细胞系侵袭行为的影响，以及对照/模拟和SV（混乱载体）的影响。目标1 c。评估p-MMP-2单独或联合照射对体外和体内脑血管生成的影响。目标1 d。确定p-MMP-2和单独或联合照射对裸鼠脑内注射的预先建立的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。具体目标2。确定p-MMP-2和辐照单独或联合作用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2。与对照/模拟和p-SV对照相比，研究p-MMP-2和照射单独或联合对胶质母细胞瘤细胞系粘附和迁移的分子机制的影响。目标2 b。与模拟和p-SV相比，确定p-MMP-2和辐照单独或联合作用对胶质母细胞瘤细胞系增殖的分子机制的影响。2 c目标。与模拟和p-SV相比，评估p-MMP-2和照射单独或联合对胶质母细胞瘤细胞系凋亡的分子机制的影响。拟议的研究将对MMP-2抑制后辐射诱导肿瘤改变的发病机制产生重要见解，进而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：尽管采取了许多治疗多形性胶质母细胞瘤的策略，但这种侵袭性脑恶性肿瘤患者的存活率仍然很低。25年来，参与野放射治疗一直是治疗胶质母细胞瘤最有效的方法。然而，越来越多的长期幸存者在这种治疗中出现了继发性副作用，这凸显了开发新的治疗方法的必要性。该建议代表了一种使用p-MMP-2 siRNA的联合治疗方法。这种策略可以改善胶质母细胞瘤的放疗效果。

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.