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性肿瘤，具有获得转移表型的倾向。这种获得的分子基础还不是很清楚，晚期这种肿瘤的治疗方法也很少。最近，我们报道了一种与众不同的基因 与非转移细胞相比，在高度转移的黑色素瘤细胞中表达的是凝血酶受体PAR-1，它通过包括血管生成、细胞信号和黏附在内的多种机制促进转移，在很大程度上是通过上调IL-8来实现的。我们的初步数据表明，转移性黑色素瘤细胞中的PAR-1被短发夹状RNA(ShRNA)沉默，抑制了其体内的生长和转移潜能。在这项拟议的研究中，我们将把这项工作推进到翻译阶段，通过在模型系统中通过包裹在中性脂质体中的siRNA来抑制PAR-1和假定毒性较低的IL-8来诱导肿瘤消退。此外，我们实验室的结果表明，血管生成因子白细胞介素8(IL-8)的表达本身通过PAR-1信号上调，与 黑色素瘤细胞的转移潜能。此外，使用全人抗IL-8的中和抗体(ABX-IL8，尚未临床使用)，我们能够在体内抑制肿瘤生长和黑色素瘤的转移。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会导致IL-8/‘nwVo的下调，从而抑制黑色素瘤的生长和转移。 我们还将评估黑色素瘤患者的治疗策略。为此，我们提出了以下具体目标： 具体目的1：评价中性脂质体包裹的PAR-1 siRNA对晚期黑色素瘤的体内疗效，作为一种可能的治疗方案。 具体目的2：评价中性脂质体包裹的IL-8 siRNA作为治疗晚期黑色素瘤和/或联合化疗的新方法的体内疗效。 具体目的3：在I期临床试验中评价IL-8 siRNA中性脂质体(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手段。

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.