Novel approach to attenuate small cell lung cancer growth and metastasis

摘要 小细胞肺癌（SCLC）是一种高度侵袭性和转移性的恶性肿瘤。不幸的是，没有一个 靶向治疗在SCLC中显示出临床活性，其5年存活率仅为6- 8%。因此 迫切需要制定针对SCLC的创新战略。近年来，microRNAs（miRs） 已显示通过沉默基因在翻译后/转录水平上在癌症中起重要作用 表情我们的初步数据显示，在小细胞肺癌细胞系以及患者中， 样品我们还观察到，在SCLC细胞系和患者中，miR-1与CXCR 4呈负相关。 样品CXCR 4/CXCL 12-信号传导轴已被探索为多种肿瘤的主要转移性信号传导轴。 癌症包括肺癌。然而，关于miR-1在调节CXCR 4中的作用， SCLC。基于我们的初步数据，我们的中心假设是miR-1的下调， 通过上调CXCR 4促进SCLC生长和转移。在本申请中，我们建议 开发用于双重靶向聚合物CXCR 4拮抗剂（PCX）纳米颗粒（NP）的纳米制剂平台 含有用于靶向SCLC的miR-1模拟物。我们将追求三个具体目标来检验我们的假设。目标1将 重点是使用细胞系和患者样本在SCLC中建立功能性miR-1和CXCR 4轴。这些 这些研究将帮助我们确定miR-1下调和CXCR 4上调在早期乳腺癌中的新作用。 检测SCLC。此外，我们将进行全局转录组（miRNA-Seq/RNA-Seq）整合， 分析以探索SCLC中的新型miR及其靶标。目标2将侧重于优化， SCLC中PCX-miR-1 NP的表征。由于SCLC具有高度侵袭性和转移性，我们将评估 PCX-miR-1 NP是否同时恢复miR-1，阻断CXCL 12/CXCR 4轴，从而抑制 在临床前小鼠模型中观察SCLC的转移。目的3：分析PCX-1的治疗效果。 SCLC自发RP（Rbf/f; Tp 53 f/f; Rosa）小鼠模型中的miR-1 NP。经鼻感染Ad-cre、RP小鼠后， 发展出重现人类SCLC特征的肿瘤。这些肿瘤具有高度侵袭性， 因此，PCX-miR-1纳米颗粒将作为理想的基于纳米制剂的治疗方法，用于治疗转移性肿瘤。 关于SCLC此外，顺铂治疗已被证明通过增加耐药性而增强耐药性。 肺癌中的CD 133 +/CXCR 4+细胞群。在这个目标中，我们还将分析PCX-miR-1 NPs是否能增强 通过重编程癌症相关的成纤维细胞实现顺铂敏化。此外，我们将确定新的目标， 使用PCX-miR-1和顺铂处理的肿瘤样品的全局转录组分析， 总之，所提出的研究将确立PCX-miR-1 NPs作为一种新的治疗策略的临床效用 用于治疗由于疾病晚期而难以治疗的SCLC患者和/或 耐药性和复发的发展。在这个项目中产生的纳米技术配方也将有 更广泛地应用于具有低miR-1和高CXCR 4表达的其他侵袭性和转移性癌症。

Abstract Small cell lung cancer (SCLC) is highly aggressive and metastatic malignancy. Unfortunately, no single targeted therapy has shown clinical activity in SCLC and its 5-year survival rate is only 6-8%. Therefore, there is an urgent need to develop innovative strategies against SCLC. In recent years, microRNAs (miRs) have been shown to play significant roles in cancer at the post-translational/transcriptional level by silencing gene expression. Our preliminary data showed reduced expression of miR-1 in SCLC cell lines as well as patient samples. We also observed that miR-1 is inversely associated with CXCR4 in SCLC cell lines and in patient samples. CXCR4/CXCL12-signaling axis has been explored as a major metastatic signaling axis for various cancers including lung cancer. However, not much is known about the role of miR-1 in regulating CXCR4 in SCLC. Our central hypothesis, based on our preliminary data, is that downregulation of miR-1 contributes to SCLC growth and metastasis by upregulating CXCR4. In this application, we propose to develop nanoformulation platform for dual-targeting polymeric CXCR4 antagonist (PCX) nanoparticles (NPs) containing miR-1 mimics for targeting SCLC. We will pursue three specific aims to test our hypothesis. Aim 1 will focus to establish functional miR-1 and CXCR4 axis in SCLC using cell lines and patient samples. These studies will help us to define the novel role of miR-1 downregulation and CXCR4 upregulation for the early detection of SCLC. Additionally, we will perform global transcriptome (miRNA-Seq/RNA-Seq) integrative analysis to explore novel miRs and their targets in SCLC. Aim 2 will be focused on optimization and characterization of PCX-miR-1NPs in SCLC. Since SCLC is highly aggressive and metastatic, we will evaluate whether PCX-miR-1 NPs simultaneously restore miR-1, block the CXCL12/CXCR4 axis, and thereby inhibit metastasis of SCLC in preclinical mouse models. Aim 3 will be designed to analyze therapeutic efficacy of PCX- miR-1NPs in SCLC spontaneous RP (Rbf/f; Tp53f/f; Rosa) mouse model. Upon nasal infection of Ad-cre, RP mice develop tumors that recapitulate human SCLC characteristics. These tumors are highly aggressive and metastatic, and therefore, PCX-miR-1 NPs will serve as ideal nanoformulation-based therapy for the management of SCLC. In addition, cisplatin treatment has been shown to enhance drug resistance through increase CD133+/CXCR4+ cell populations in lung cancer. In this aim, we will also analyze if PCX-miR-1 NPs enhance cisplatin sensitization by reprogramming cancer-associated fibroblasts. Additionally, we will identify novel targets of miR-1 for SCLC using global transcriptome analysis of PCX-miR-1 and cisplatin treated tumor samples. Altogether, the proposed studies will establish the clinical utility of PCX-miR-1 NPs as a novel therapeutic strategy for the treatment of SCLC patients who are difficult to treat due to their advanced disease stage and/or development of drug resistance and recurrence. Nanotechnogy formulation generated in this project will also have broader application in other aggressive and metastatic cancers that have low miR-1 and high CXCR4 expression.