IL-8 targeted Nanotherapy for Prostate Cancer.

描述(申请人提供)：R21探索性研究基金的修订申请是对PA-11-149题为“生物和医学中的纳米科学和纳米技术”的回应。该项目涉及国家癌症研究所感兴趣的一个领域，该领域寻求对基于纳米技术的方法进行评估，以实现对转移的理解、预防、检测和消除。在这一探索性研究项目中，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(BDT IL-8-siRNA纳米复合体)在CAP细胞中有效和持久地下调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网络，用于CAP的活体动物模型。BDT IL-8-siRNA纳米复合体构建在烯丙基功能化的L-丙交酯单体上，产生官能化的聚乳酸主链，具有高度的生物相容性，使其成为纳米治疗的理想试剂。尽管我们之前对PC-3帽细胞系进行了初步研究，但在本方案中我们将使用LNCaP细胞系，因为它表达前列腺特异性 膜抗原(PSMA)允许其通过含有针对PSMA的单抗(MAb)的定制纳米网络来选择性靶向。首先，我们将进行体外实验，以确定BDT IL-8-siRNA纳米网络处理LNCaP细胞的效果。Nanopex Controls将整合一种杂乱的siRNA。随后的体内实验将使用我们建立的裸鼠裸鼠模型，将LNCaP细胞原位接种到前列腺癌中，以形成原发肿瘤和随后的转移。动物将接受BDT IL-8-siRNA纳米复合体的治疗，直接静脉注射到前列腺中。此外，BDT IL-8-siRNA纳米复合体将偶联MAb到PSMA，用于体内系统靶向递送。治疗的动物将被前瞻性地跟踪，我们预计与对照组相比，肿瘤和转移的消退。将对血清和肿瘤组织进行分析，以抑制治疗动物的IL-8表达，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纳米网络在CAP患者中的最终临床试验建立概念证据。

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.