Albumin-binding nanocomplexes for delivery of combination siRNA therapeutics to KRAS-driven cholangiocarcinoma

白蛋白结合纳米复合物用于向 KRAS 驱动的胆管癌递送组合 siRNA 疗法

基本信息

批准号：
10389971
负责人：
Justin Han-Je Lo
金额：
$ 7.62万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10389971
关键词：
Address Adverse effects Albumins Apoptotic Binding Biological Biological Availability Biological Response Modifier Therapy Blood Circulation Blood Tests Cell Death Cell Line Chemicals Chemistry Chloroquine Cholangiocarcinoma Clinical Diagnosis Disease Drug Kinetics Endosomes Extrahepatic FDA approved FRAP1 gene Face Fatty Acids Gene Combinations Gene Silencing Gene Targeting Generations Genes Goals Hepatic Histology Impairment In Vitro Induction of Apoptosis Inhibition of Cell Proliferation Intrahepatic Cholangiocarcinoma Intravenous K-ras mouse model KRAS oncogenesis KRAS2 gene Kidney MAP Kinase Gene MCL1 gene Malignant Neoplasms Mediating Modeling Molecular Molecular Target Monitor Mutate Mutation Oncogenes Oncogenic Organ Outcome Pathway interactions Patients Penetration Pharmaceutical Preparations Prognosis Property Quantitative Reverse Transcriptase PCR RNA delivery RNA-targeting therapy Reporter Genes Research Resistance Resistance development Signal Transduction Site Small Interfering RNA Structure Tail Technology Testing Therapeutic Therapeutic Effect Therapeutic Use Study Time Tumor Tissue Twin Multiple Birth Unresectable Western Blotting Work base chemotherapy density design experimental study immunogenicity improved in vivo intrahepatic knock-down mTOR Signaling Pathway molecular targeted therapies monolayer mouse model mutant nanocomplexes new therapeutic target next generation nuclease pre-clinical preempt response small molecule small molecule therapeutics standard of care subcutaneous targeted treatment therapeutic RNA therapy outcome therapy resistant tissue culture tumor uptake

项目摘要

Research Summary Patients diagnosed with advanced cholangiocarcinoma (CCA) have poor overall outcomes and face limited treatment options. Small interfering RNA (siRNA) therapeutics offer an attractive strategy for silencing oncogenic drivers of CCA that lack FDA-approved molecularly-targeted therapeutics, particularly KRAS, which is mutated in ~22% of intrahepatic and ~42% of extrahepatic CCAs. Combination siRNA therapeutics may be devised to preempt compensatory resistance pathways that frequently arise in the course of targeted therapy. Furthermore, siRNA may be targeted to sites of malignancy, avoiding adverse effects due to molecularly on- target activity in healthy organs, as is seen with many small molecule drugs. However, delivery challenges including nuclease degradation, rapid clearance, and lack of a mechanism for cellular uptake or endosome escape have traditionally limited clinical use of siRNA. Our group has recently developed technology to chemically modify siRNAs with twin fatty acids (siRNA-L2) that form non-covalent nanocomplexes with endogenous albumin (alb-NCs). Since albumin is ordinarily long-circulating in the vasculature but is actively taken up by tumors, we have found that alb-NCs extend siRNA circulation time, promote homogeneous tumor penetration, and increase tumor-selective siRNA uptake. Such a technology is well-suited for fibrotic tumors like CCA, for which an active uptake mechanism is necessary for delivery of large biologic therapeutics. However, the siRNA-L2 design has not yet been integrated with “on-board” functionality to escape from endosomes, which represent a critical barrier to siRNA activity. In this project, I propose to optimize and tailor the siRNA-L2 alb-NC platform to enable efficient tumor-selective knockdown of oncogenic drivers in CCA. The studies proposed here will test the hypothesis that alb-NC-mediated delivery of a combination of siRNAs targeting both KRAS and complementary resistance pathways will provide therapeutic benefit in KRAS- driven cholangiocarcinoma. First, I will chemically optimize alb-NCs for efficient endosomal escape and intracellular siRNA delivery. Second, I will credential gene targeting of KRAS in combination with rationally- selected complementary gene targets involved in mTOR signaling or apoptotic pathways implicated in KRAS treatment resistance. I will then perform pre-clinical therapeutic studies using alb-NCs to deliver KRAS-based siRNA combinations to orthotopic mouse models of KRAS-mutant intrahepatic CCA. This project will thus address a pressing need for new targeted therapeutic approaches in CCA by developing efficacious siRNA delivery technologies to target key gene combinations in KRAS-driven disease.

研究总结诊断为晚期胆管癌 (CCA) 的患者总体预后较差，并面临小干扰 RNA (siRNA) 疗法提供了一种有吸引力的沉默策略。 CCA 的致癌驱动因素缺乏 FDA 批准的分子靶向治疗药物，特别是 KRAS，约 22% 的肝内 CCA 和约 42% 的肝外 CCA 发生突变组合 siRNA 治疗可能是有效的。旨在抢占靶向治疗过程中经常出现的代偿性阻力途径。此外，siRNA 可以靶向恶性肿瘤部位，避免由于分子作用而产生的不利影响。正如许多小分子药物所见，目标是健康器官的活性。然而，递送面临挑战。包括核酸酶降解、快速清除以及缺乏细胞摄取或内体机制传统上，siRNA 的临床应用受到限制，我们的团队最近开发了技术用双脂肪酸 (siRNA-L2) 对 siRNA 进行化学修饰，形成非共价纳米复合物内源性白蛋白（alb-NCs），因为白蛋白通常在脉管系统中长期循环，但活跃。被肿瘤吸收后，我们发现alb-NCs延长了siRNA的循环时间，促进肿瘤同质化渗透，并增加肿瘤选择性 siRNA 的吸收，这种技术非常适合纤维化肿瘤，例如纤维化肿瘤。 CCA，对于大的生物治疗药物的输送来说，主动摄取机制是必需的。 siRNA-L2 设计尚未与“机载”功能集成以逃离核内体，这代表了 siRNA 活性的一个关键障碍，在这个项目中，我建议优化和定制 siRNA-L2 alb-NC。平台能够有效地对 CCA 中的致癌驱动因素进行肿瘤选择性敲除。这里提出的研究将检验以下假设：alb-NC 介导的组合递送针对 KRAS 和互补耐药途径的 siRNA 将为 KRAS 提供治疗益处首先，我将对 alb-NC 进行化学优化，以实现有效的内体逃逸和其次，我将证明 KRAS 的基因靶向与合理的结合。选定参与 mTOR 信号传导或 KRAS 涉及的细胞凋亡途径的互补基因靶标然后我将使用 alb-NC 进行基于 KRAS 的临床前治疗研究。因此，该项目将与 KRAS 突变肝内 CCA 的原位小鼠模型进行 siRNA 组合。通过开发有效的 siRNA 满足 CCA 对新靶向治疗方法的迫切需求针对 KRAS 驱动疾病中关键基因组合的递送技术。