An Integrated Nano-Cell Delivery Platform of Theranostics for Lung Cancers

描述（由申请人提供）： 这项研究提案的主要目标是开发一种新的方法来检测和治疗肺癌，该方法利用特殊工程化的壳聚糖治疗诊断纳米颗粒（CTN），并使用肺靶向支持细胞（SC）来递送携带癌细胞检测部分和基因或药物的纳米颗粒，以有效治疗肺癌。在全世界范围内，估计有130万例肺癌病例，其中近100万人将在同一年死亡。这一惊人的死亡率超过了主要类型癌症（乳腺癌，前列腺癌和结肠癌）的死亡人数总和，占美国所有死亡人数的6%。肺癌患者通常表现为局部晚期或播散性疾病。大约十分之六的肺癌患者在诊断后一年内死亡。主要的局限性是我们无法早期发现癌症或缓解癌症，以及缺乏专门治疗癌细胞而不是正常健康细胞的药物。使用靶向纳米颗粒技术研究了药物向肺部的递送，该技术使用天然的、生物相容性的和可生物降解的壳聚糖纳米颗粒和靶向心房利钠素受体A的短干扰RNA，心房利钠素受体A是一种新型抗癌靶点，在2008年被指定为肿瘤学中用于癌症的“领先发现”。然而，壳聚糖纳米颗粒尚未用于治疗诊断研究。最近，我们已经发现，当睾丸外的SC装载有纳米颗粒（即，SNAP）并静脉注射到小鼠中，通过循环直接行进到肺的毛细血管床，在那里它们释放它们的纳米颗粒货物。SNAP方法将治疗剂向肺的递送从当前标准的约20%增加至高达90%。这些发现共同导致了这样的假设，即用包含近红外染料和siNPRA的癌细胞靶向治疗诊断纳米颗粒包装的SC可以特异性地递送至肺，这可以提供肺癌的检测和治疗。为了检验这些假设，提出了以下具体目标。目标#1开发用于肺癌的聚合物治疗诊断剂。在这一目标中，计划制定策略，以开发明确的特殊工程球形聚合物纳米颗粒（100- 300 nm），并通过细胞载体递送到肺中，用于诊断（通过体内成像和MRI）和治疗目的。目标2。优化并进一步开发Theranostic SNAP的稳健的肺特异性递送。在这个目标中，计划通过调节作为报告基因的增强型绿色荧光蛋白（EGFP）的细胞内表达来优化递送系统。将评估小鼠肺中基因表达的递送效率、时间过程、剂量反应和持久性。此外，建议通过评估SC分泌产物在使用或不使用纳米颗粒递送后在肺中的表达来评估SNAP方法的短期和长期安全性。目标3。在肺转移的LLC 1模型中测试细胞靶向SNAP方法并验证siNPRA的治疗作用。在这项研究中，建议使用包装有siNPRA或psiNPRA的癌细胞靶向多功能治疗诊断剂来评估SNAP递送系统在预防LLC 1肺转移中的功效。目标4。评估SNAP-iNPRA在自发性肺癌的CC 10 Cre-K-Ras-G12 D模型中的功效。为此，计划通过使LSL-K-ras-G12 D小鼠与CC 10-Crig小鼠交配以产生CC 10-Cre-LSL-K-rasG 12 D小鼠来建立自发性肺癌模型，并在小鼠模型中评估与诱导型siNPRA或psiNPRA复合的治疗诊断剂在抑制由K-ras过表达诱导的自发性肺癌中的功效。调查人员可以使用所有试剂和方法，并已开展必要的合作和可行性研究。这些研究的结果有望验证NPRA作为肺癌的治疗靶点，以及SNAP基因递送方法作为治疗肺癌的安全有效方法的实用性，并为利用SNAP治疗转移性肺癌的临床试验奠定基础。

DESCRIPTION (provided by applicant): The primary goal of this research proposal is to develop a novel approach to detect and treat lung cancers that utilizes specially engineered chitosan theranostic nanoparticles (CTNs) and uses lung-targeting Sertoli cells (SCs) to deliver nanoparticles carrying a cancer cell detecting moiety and a gene or drug to effectively treat lung cancers. Worldwide, lung cancer affects an estimated 1.3 million cases of which close to a million will die the same year. This staggering death rate exceeds the combined number of deaths from the leading types of carcinoma (breast, prostate and colon cancer), and accounts for 6% of all deaths within the US. Lung cancer patients often present with locally advanced or disseminated disease. About 6 out of 10 people with lung cancer die within 1 year of diagnosis. The major limitations have been our inability to detect cancers or remission of cancers early and the lack of drugs, which specifically treat cancer cells and not normal healthy cells. The drug delivery to the lung has been investigated using targeted nanoparticle technology that uses natural, biocompatible and biodegradable chitosan nanoparticles and short interference RNA targeting the atrialnatriuretic receptor A, a novel anti-cancer target that was designated a 'lead discovery' in oncology in 2008 for cancers. However, chitosan nanoparticles have not been used for theranostic studies. Recently, we have found that extra-testicular SCs, when loaded with nanoparticles (i.e., SNAPs) and injected intravenously into mice, travel via the circulation directly to the capillary bed of the lung where they release their nanoparticle cargo. The SNAP method increases delivery of therapeutics to the lung from the current standard of about 20% up to 90%. Together these findings have led to the hypothesize that SCs packaged with cancer cell-targeting theranostic nanoparticles comprising a near-infrared dye and siNPRA can be delivered specifically to the lung, which may provide both detection and treatment of lung cancers. To test these hypotheses, the following specific aims are proposed. Aim #1 Develop a polymer theranostics for lung cancers. In this aim, it is planned to develop strategy to develop well-defined specially engineered spherical polymeric nanoparticles (100-300nm) and deliver into lung via cell carriers for diagnostic (by in vivo imaging and MRI) and therapeutic purposes. Aim #2. Optimize and further develop a robust lung-specific delivery of Theranostic SNAPs. In this aim it is planned to optimize the delivery system by regulating intracellular expression of the enhanced green fluorescent protein (EGFP) as a reporter. The delivery efficiency, time course, dose response and persistence of gene expression in the mouse lung will be evaluated. In addition, it is proposed to evaluate the short- and long-term safety of the SNAP method by assessing the expression of SC secretory products in the lung following their delivery with or without nanoparticles. Aim #3. Test cell-targeted SNAP method in LLC1 model of lung metastasis and validate therapeutic role of siNPRA. In this study, it is proposed to evaluate the efficacy of the SNAP delivery system using cancer cell-targeted multifunctional theranostics packaged with siNPRA or psiNPRA in preventing LLC1 lung metastasis. Aim #4. Evaluate the efficacy of SNAP-iNPRA in a CC10 Cre- K-Ras-G12D model of spontaneous lung cancer. In this aim, it is planned to establish a model of spontaneous lung cancer by breeding LSL-K-ras- G12D mice with CC10- Cretg mice to generate CC10-Cre-LSL-K-ras G12D mice and to evaluate the efficacy of theranostics complexed with the inducible siNPRA or psiNPRA in suppressing spontaneous lung cancer induced by overexpression of K-ras in a mouse model. The investigators have access to all reagents and methods and have already developed necessary collaborations and feasibility studies. The results of the proposed studies are expected to validate NPRA as a therapeutic target for lung cancer and the utility of the SNAP method of gene delivery as a safe and effective approach to treat lung cancers, and set the stage for clinical trials for utilizing SNAPs to treat metastatic lung cancers.