In vivo RNAi nanoparticle cancer gene function reagent

描述（由申请人提供）：尽管最近取得了进展，但癌症仍然是一个主要杀手，迫切需要更有效的治疗方法，不幸的是，对于转移性非小细胞肺癌（NSCLC）尤其如此，这种癌症折磨着极其多的患者，是最难治疗的癌症之一。随着靶向治疗的日益成功，对非小细胞肺癌生物化学和病理的进一步了解是确定新的和更好的治疗方法的关键瓶颈。为此，人们正在努力开发适用于临床样本的先进基因组学和蛋白质组学方法，其中一项重大进展来自使用激光显微解剖和利用大型福尔马林固定组织切片库进行蛋白质提取和分析的能力。然而，利用这些极有价值的信息开发靶向治疗的一个主要障碍是表征肿瘤相关蛋白的功能作用。另一个有助于解决这一需求的重大进展是RNA干扰（RNAi）的最新发展，但这种表征基因功能的革命性能力主要局限于细胞培养研究，在动物疾病模型（如非小细胞肺癌异种移植肿瘤模型）中的体内给药方法有限，而且没有可用的研究试剂。我们的假设是，我们可以采用我们的阳离子多肽核酸纳米颗粒技术，我们已经发现该技术有效地将RNAi药物体内递送到原发性乳腺癌异种移植模型中，用于一组转移性和原发性NSCLC异种移植肿瘤模型。计划中的研究旨在确定一种可商业化的试剂在体内应用于转移性和原发性非小细胞肺癌异种移植肿瘤模型的可行性，其I期目标如下：1)建立两对转移性和原发性非小细胞肺癌异种移植模型；2)使用阳性对照RNAi剂对HK多肽和纳米颗粒结构进行调整和优化，以便在这些模型中进行体内递送；3)使用一组在转移性肿瘤或原发肿瘤病变中差异表达的候选靶标，评估优化后的HK RNAi纳米颗粒用于靶标验证的有效性，这些靶标均来自患者组织样本。如果成功，II期将进行体内RNAi试剂的商业化开发，用于非小细胞肺癌异种移植模型基因功能靶标验证，包括转移性癌症。公共卫生相关性：转移性非小细胞肺癌（NSCLC）困扰着大量患者，是最难治疗的癌症之一，因此是应用基因组和蛋白质组学方法的研究进展的主要领域，目的是确定更好的药物靶点。然而，仍然存在的一个主要障碍是需要商业试剂来直接在转移性非小细胞肺癌动物模型中使用RNA干扰（RNAi）。表征基因功能和鉴定候选药物靶点的RNAi革命性能力主要局限于细胞培养研究。我们计划的研究是将我们的阳离子多肽核酸纳米颗粒技术应用于一组转移性和原发性非小细胞肺癌异种移植肿瘤模型的体内递送RNAi药物。如果成功，该试剂有望在了解这种威胁生命的疾病方面取得快速进展，为患者提供更好的靶向治疗选择。

DESCRIPTION (provided by applicant): In spite of recent advances, cancer remains a major killer posing a dire need for more effective therapy, and unfortunately this is particularly true for metastatic non-small cell lung cancer (NSCLC), which afflicts an extremely large number of patients and is one of the least treatable cancers. With the growing success of targeted therapeutics, it has become clear that a greater understanding of NSCLC biochemistry and pathology is a crucial bottleneck in identifying new and better treatments. To that end considerable efforts are underway developing advanced genomic and proteomic methods applicable to clinical samples, with a major advance coming from a capability for using laser micro-dissection and protein extraction and analysis with the large libraries of formalin fixed tissue sections. However, a remaining major barrier to using this extremely valuable information for development of targeted therapeutics is to characterize the functional role of the tumor associated proteins. Yet another major advance helping address this need is the recent development of RNA interference (RNAi) but this revolutionary capability to characterize gene function has been confined largely to cell culture studies, with limited methods for in vivo administration in animal disease models such as NSCLC xenograft tumor models, and none available as a research reagent. Our hypothesis is that we can adapt our cationic polypeptide nucleic acid nanoparticle technology, which we have found effective for in vivo delivery of RNAi agents to a primary breast cancer xenograft model, for a set of metastatic and primary NSCLC xenograft tumor models. The planned research to determine feasibility of a commercializable reagent for in vivo RNAi application to metastatic and primary NSCLC xenograft tumor models has the following objectives for Phase I: 1) establish two pair of metastatic and primary NSCLC xenograft models, 2) adapt and optimize the HK polypeptide and nanoparticle structure for in vivo delivery with these models using a positive control RNAi agent, and 3) evaluate the effectiveness of the optimized HK RNAi nanoparticle for target validation using a set of candidate targets found to be differentially expressed in metastatic tumor or primary tumor lesions but not both from patient tissue samples. If successful, Phase II will undertake commercial development of a reagent for in vivo RNAi for NSCLC xenograft model gene function target validation with capabilities including metastatic cancer. PUBLIC HEALTH RELEVANCE: Metastatic non-small cell lung cancer (NSCLC) afflicts an extremely large number of patients and is one of the least treatable cancers and thus is a major area for research applying advances in genomic and proteomic methods with the objective to identify better drug targets. However, a remaining major barrier is a need for commercial reagents for using RNA interference (RNAi) directly in animal models of metastatic NSCLC. The RNAi revolutionary capability to characterize gene function and identify candidate drug targets has been confined largely to cell culture studies. Our planned studies are to adapt our cationic polypeptide nucleic acid nanoparticle technology for in vivo delivery of RNAi agents for a set of metastatic and primary NSCLC xenograft tumor models. If successful, the resulting reagent is expect to enable rapid advances in understanding this life threatening disease that produce better targeted therapeutic treatment options for patients.