Targeting Angiogenesis and the Tumor Microenvironment Utilizing si RNA

2.项目总结（见说明）： 黑色素瘤是一种潜在的致命肿瘤，具有获得转移表型的倾向。这种收购的分子基础是不是很好地定义和几个治疗方式可用于这种肿瘤在先进的阶段。最近，我们报道了一个基因， 与非转移性细胞相比，在高度转移性黑素瘤细胞中表达的是凝血酶受体PAR-1，其通过多种机制促进转移，所述机制包括血管生成、细胞信号传导和粘附，在很大程度上通过IL-8的上调。我们的初步数据表明，PAR-1沉默短发夹RNA（shRNA）在转移性黑色素瘤细胞抑制其生长和转移潜力在体内。在这项拟议的研究中，我们将推进这项工作的翻译阶段，追求PAR-1的抑制，以及假定毒性较低的IL-8通过siRNA封装在中性脂质体中诱导肿瘤消退模型系统。此外，来自我们实验室的结果已经证明，血管生成因子白细胞介素8（IL-8）的表达，其本身通过PAR-1信号传导上调， 黑色素瘤细胞的转移潜能此外，使用针对IL-8的完全人中和抗体（ABX-IL 8，其不能用于临床使用），我们能够在体内抑制黑素瘤的肿瘤生长和转移。我们的假设是，静脉注射IL-8小干扰RNA（siRNA） 包装在中性脂质体中的IL-8/nwVo将引起IL-8/nwVo的下调，从而抑制黑素瘤生长和转移。 我们还将评估黑色素瘤患者的治疗策略。为此，我们提出以下具体目标： 具体目标1：评价中性脂质体包裹PAR-1 siRNA单独或联合化疗治疗晚期黑色素瘤的体内效应。 具体目标二：评价中性脂质体包裹的IL-8 siRNA作为一种新的治疗模式单独和/或与化疗联合治疗晚期黑色素瘤的体内效果。 具体目的3：在I期临床试验中评估中性脂质体包装的IL-8 siRNA（IL-8 siRNA-DOPC）的安全性和有效性。 这是首次尝试使用纳米技术作为晚期黑色素瘤的治疗方式。

2. PROJECT SUMMARY (See instructions): Melanoma is a potentially lethal neoplasm with, a propensity for acquiring the metastatic phenotype. The molecular basis for this acquisition is not very well defined and few treatment modalities are available for this neoplasm at advanced stages. Recently, we reported that one of the genes that stands out as differentially expressed in highly, metastatic melanoma cells as compared to non-metastatic cells, is the thrombin receptor PAR-1, which promotes metastases through multiple mechanisms including angiogenesis, cell signaling, and adhesion, in large part via upregulation of IL-8. Our preliminary data indicated that PAR-1 silencing by short hairpin'RNA (shRNA) in.metastatic melanoma cells inhibited their growth and metastatic potential in vivo. In this proposed study, we will advance this work to the translational phase by pursuing inhibition of PAR-1 as well as the presumed less toxic IL-8 via siRNA encapsulated in neutral liposomes to induce tumor regression in model systems. In addition, results from oUr laboratory have demonstrated that expression of the angiogenic factor interieukin 8 (IL-8), which is itself upregulated through PAR-1 signaling, correlates with the metastatic potential of melanoma cells. Moreover, using a fully human neutralizing antibody against IL-8 (ABX-IL8 which is not available for clinical use), we were able to inhibit tumor growth and metastasis of melanoma in vivo. Our hypothesis is that intravenous administration of IL-8 small interfering RNA (siRNA) packaged in neutral liposomes will cause downregulation of IL-8 /'nwVo, thereby inhibiting melanoma growth and metastasis. We will also evaluate the therapeutic strategy in melanoma patients. To these ends, we propose the following Specific Aims: Specific Aim 1: To evaluate the iri vivo effects of PAR-1 siRNA pacltaged in neutral liposomes as a possible therapeutic modality for advanced melanoma alone and/or in combination with chemotherapy. Specific Aim 2: To evaluate the in vivo effects of IL-8 siRNA packaged in neutral liposomes as a new therapeutic modality for advanced melanoma alone and/or in combination with chemotherapy. Specific Aim 3: To evaluate the safety and efficacy of IL-8 siRNA packaged in neutral liposomes (IL-8 siRNA-DOPC) in a Phase I clinical trial. This is the first attempt at using nanotechnology as a therapeutic modality for advanced melanoma.