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，没有单曲 靶向治疗在小细胞肺癌中已显示出临床活性，其5年生存率仅为6-8%。因此，在那里 迫切需要制定针对小细胞肺癌的创新战略。近年来，microRNAs(MiRs)已经 已被证明在翻译后/转录水平上通过沉默基因在癌症中发挥重要作用 表情。我们的初步数据显示，miR-1在小细胞肺癌细胞系和患者中的表达减少 样本。我们还观察到，在小细胞肺癌细胞系和患者中，miR-1与CXCR4呈负相关 样本。CXCR4/CXCL12-信号轴被认为是多种疾病的主要转移性信号轴 包括肺癌在内的癌症。然而，关于miR-1在调控CXCR4中的作用还知之甚少 SCLC。根据我们的初步数据，我们的中心假设是miR-1的下调 通过上调CXCR4促进小细胞肺癌的生长和转移。在本申请中，我们建议 开发双靶向聚合物CXCR4拮抗剂纳米粒(NPs)的纳米制剂平台 含有针对小细胞肺癌的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 重点利用细胞系和患者样本建立小细胞肺癌的miR-1和CXCR4功能轴。这些 研究将帮助我们确定miR-1下调和CXCR4上调在早期的新作用 小细胞肺癌的检测。此外，我们还将进行全局转录组(miRNA-Seq/RNA-Seq)的整合 分析小细胞肺癌中新的MIR及其靶点。目标2将专注于优化和 小细胞肺癌中PCX-miR-1NPs的特征由于小细胞肺癌具有高度侵袭性和转移性，我们将对 PCX-miR-1 NPs是否同时恢复miR-1，阻断CXCL12/CXCR4轴，从而抑制 小细胞肺癌在临床前期小鼠模型中的转移AIM 3将用于分析PCX的治疗效果- 小细胞肺癌自发性RP(RBF/f；Tp53f/f；ROSA)小鼠模型中的MIR-1NPs。Ad-cre、RP小鼠鼻腔感染的实验研究 发展具有人类小细胞肺癌特征的肿瘤。这些肿瘤侵袭性很强， 转移性，因此，PCX-miR-1纳米粒将作为理想的基于纳米制剂的治疗方法 小细胞肺癌的研究。此外，顺铂治疗已被证明通过增加耐药性来增强耐药性 CD133+/CXCR4+细胞在肺癌中的表达为此，我们还将分析PCX-miR-1 NPs是否会增强 顺铂通过对癌症相关成纤维细胞重新编程来增敏。此外，我们将确定新的目标 通过对PCX-miR-1和顺铂治疗的肿瘤样本进行整体转录组学分析，对小细胞肺癌的miR-1进行了研究。 总之，拟议的研究将确立PCX-miR-1纳米粒作为一种新的治疗策略的临床实用价值。 用于治疗因疾病晚期和/或难以治疗的小细胞肺癌患者 耐药和复发的发展。在这个项目中产生的纳米技术配方也将具有 更广泛地应用于其他miR-1低表达和CXCR4高表达的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.