IL-8 targeted Nanotherapy for Prostate Cancer.

描述（由申请人提供）：这一修订后的申请R21探索性研究补助金是为了响应PA-11-149题为“纳米科学和纳米技术在生物学和医学”。该项目涉及国家癌症研究所的一个感兴趣的领域，该领域寻求评估基于纳米技术的方法，以了解，预防，检测和消除转移。在这个探索性的研究项目中，我们将验证我们最近开发的，独特的，可生物降解的靶向纳米颗粒与IL-8特异性siRNA复合物（BDT IL- 8-siRNA nanoplex）在CaP细胞中有效和长期敲低IL-8基因表达，导致其死亡。该项目的一个组成部分将使我们能够生产足够数量的治疗性纳米复合物，用于CaP的体内动物模型。BDT IL-8-siRNA纳米复合物构建在烯丙基官能化的L-丙交酯单体上，产生官能化的聚乳酸（PLA）骨架，并且具有高度生物相容性，使其成为纳米治疗的理想试剂。尽管我们先前用PC-3 CaP细胞系进行了初步研究，但我们将在本建议中使用LNCaP细胞系，因为它表达前列腺特异性的前列 膜抗原（PSMA），允许其通过掺入针对PSMA的单克隆抗体（MAb）的定制纳米复合物进行选择性靶向。首先，我们将进行体外实验以确定用BDT IL-8-siRNA纳米复合物处理LNCaP细胞的效果。Nanoplex对照将掺入乱序siRNA。随后的体内实验将使用我们建立的人CaP的无胸腺、裸小鼠模型，采用将LNCaP细胞原位接种到前列腺中，允许形成原发性肿瘤和随后的转移。将用直接静脉内施用到前列腺中的BDT IL-8-siRNA纳米复合物处理动物。此外，将施用偶联至PSMA的MAb的BDT IL-8-siRNA纳米复合物用于体内全身靶向递送。将前瞻性随访治疗动物，我们预计与对照组相比，肿瘤和转移灶消退。将分析血清和肿瘤组织中IL-8表达的敲低以及血管生成和抗凋亡因子的表达。这些探索性研究旨在为BDT IL-8-siRNA纳米复合物在CaP患者中的最终临床试验建立概念证明。

DESCRIPTION (provided by applicant): This revised application for an R21 exploratory research grant is in response to PA-11-149 entitled "Nanoscience and Nanotechnology in Biology and Medicine". This project addresses an area of interest of the National Cancer Institute seeking evaluation of nanotechnology-based methods to enable the understanding, prevention, detection, and elimination of metastases. In this exploratory research project we shall validate our recently developed, unique, biodegradable targeted nanoparticle complexed with IL-8 specific siRNA (BDT IL- 8-siRNA nanoplex) for the efficient and prolonged knockdown IL-8 gene expression in CaP cells resulting in their death. A component of this project will allow us to produce sufficient quantities of therapeutic nanoplexes for use in an in vivo animal model of CaP. BDT IL-8-siRNA nanoplexes are built on allyl-functionalized L-lactide monomers, yielding a functionalized polylactic acid (PLA) backbone and are highly biocompatible, making them ideal agents for nanotherapy. In spite of our prior preliminary studies with the PC-3 CaP cell line, we shall use the LNCaP cell line in this proposal because it expresses prostate specific membrane antigen (PSMA) allowing for its selective targeting by customized nanoplexes incorporating monoclonal antibodies (MAb) to PSMA. Initially we shall perform in vitro experiments to determine the effect of treating LNCaP cells with BDT IL-8-siRNA nanoplexes. Nanoplex controls will incorporate a scrambled siRNA. Subsequent in vivo experiments will use our established athymic, nude murine model of human CaP employing orthopic inoculation of LNCaP cells into the prostate allowing formation of a primary tumor and subsequent metastases. Animals will be treated with BDT IL-8-siRNA nanoplexes administered i.v directly into the prostate. Further, BDT IL-8-siRNA nanoplexes coupled to MAb to PSMA will be administered for systemic targeted delivery in vivo. Treated animals will be followed prospectively and we expect to see regression of tumors and metastases compared to controls. Serum and tumor tissue will be analyzed for knockdown of IL-8 expression in treated animals as well as the expression of angiogenic and anti-apoptotic factors. These exploratory studies are designed to establish proof of concept for eventual clinical trials of BDT IL-8-siRNA nanoplexes in patients with CaP.