Therapeutic Targeting of the Ras Pathway By Nanoparticle Delivery of siRNA

通过 siRNA 纳米颗粒递送来治疗 Ras 通路

• 批准号: 8540374
• 负责人: Leaf Huang
• 金额: $ 14.43万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8540374
• 关键词: Biodistribution; Cancer Center; Cancer Etiology; Cancer cell line; Cessation of life; Collaborations; DNA; Development; Dose; Drug Formulations; Drug Kinetics; EGFR inhibition; Encapsulated; Endosomes; Epidermal Growth Factor Receptor; Gene Expression; Gene Silencing; Genetic; Genetically Engineered Mouse; Human; Image; In Vitro; Injection of therapeutic agent; Lesion; Lipids; Liposomes; Liver; Luciferases; Lung Adenocarcinoma; Lung Neoplasms; MEKs; Maintenance; Malignant Neoplasms; Malignant neoplasm of lung; Mediating; Membrane; Metabolism; Molecular Analysis; Molecular Weight; Mono-S; Mus; Mutation; Nanotechnology; Non-Small-Cell Lung Carcinoma; Oncogene Activation; Oncogenes; Pathway interactions; Patients; Pharmaceutical Chemistry; Pharmaceutical Preparations; Signal Transduction; Small Interfering RNA; Spleen; Structure; Surface; Swelling; Testing; Therapeutic; Tissues; Toxic effect; Treatment Efficacy; Tumor Suppressor Genes; Tyrosine Kinase Inhibitor; United States; Woman; Work; Xenograft procedure; c-Myc Staining Method; calcium phosphate; cancer cell; chemotherapy; density; design; efficacy testing; gemcitabine; improved; inorganic phosphate; knock-down; men; mortality; mouse model; mutant; nanoencapsulated; nanoparticle; neoplastic cell; novel; novel strategies; polycation; response; small molecule; standard care; therapeutic target; tumor; tumor growth; tumor xenograft; uptake

Project 2: Therapeutic Targeting of the Ras Pathway by Nanoparticle Delivery of siRNA Three types of nanoparticles, i.e. LPD, LCP and PRINT, will be used to deliver siRNA to lung cancer cells in a genetically engineered mouse (GEM) model (Kim, co-PI). LPD is a self-assembled, core/membrane nanoparticle, the unique feature of which is the high density ofthe surface grafted PEG chains. Previous work by Huang (co-PI) has shown that LPD accumulates in xenografted human lung tumors at the level of 60-80 % injected dose per g of tissue with minimal uptake by liver and spleen. Three daily i.v. injections of LPD containing siRNA effectively silenced the tumor target gene expression. LCP is similar in structure to LPD, but the core is replaced with calcium phosphate amorphous nanoprecipltate that dissolves in the endosomal acidic pH to release its cargo and swells and bursts the endosomes. The PRINT (DeSimone, co-1) are topdown manufactured nanoparticles In which any carrier and cargo materials can be readily loaded. Dr. DeSimone has shown that siRNA can be successfully loaded in PRINT and delivered to tumor cells for gene silencing. A salient feature of the project is the KRas driven lung cancer GEM. sIRNAs designed against Ras and Ras effector pathways will be tested and delivered as therapeutics. This is particularly significant, because Ras is considered an undruggable target by conventional medicinal chemistry approaches. Furthermore, gemcitabine mono (or di-) phosphate (GMP or GDP), metabolites of gemcitabine will be formulated in LCP nanoparticles and delivered to the tumor cells by itself or together with siRNA. This is a novel approach that takes advantage of the large cargo capacity of these nanoparticles. The project is milestone driven in that by the end of the year 3, at least one nanoparticle formulation will be identified for therapy, pharmacokinetics and toxicity studies In GEM. The formulation should be ready for detailed characterization and toxicity studies in collaboration with Nanotechnology Characterization Lab at NCI for further translational development

项目2：通过siRNA的纳米颗粒递送治疗性靶向Ras通路 三种类型的纳米颗粒，即LPD、LCP和PRINT，将用于在基因工程小鼠（GEM）模型中将siRNA递送至肺癌细胞（Kim，co-PI）。LPD是一种自组装的核/膜纳米粒子，其独特之处在于表面接枝了高密度的PEG链。Huang（co-PI）先前的工作表明，LPD在异种移植的人肺肿瘤中以60- 80%注射剂量/g组织的水平蓄积，肝脏和脾脏的摄取最小。每天三次静脉注射含有siRNA的LPD有效地沉默了肿瘤靶基因的表达。LCP在结构上与LPD相似，但核心被磷酸钙无定形纳米脂质体替代，磷酸钙无定形纳米脂质体溶解于内体酸性pH中以释放其货物并溶胀和破裂内体。PRINT（DeSimone，co-1）是自上而下制造的纳米颗粒，其中可以容易地装载任何载体和货物材料。DeSimone博士已经证明，siRNA可以成功地加载到PRINT中，并传递到肿瘤细胞中进行基因沉默。该项目的一个突出特点是KRas驱动的肺癌GEM。针对Ras和Ras效应子途径设计的sIRNA将被测试并作为治疗剂递送。这是特别重要的，因为Ras被认为是传统药物化学方法不可治疗的目标。 此外，吉西他滨单（或二）磷酸盐（GMP或GDP）、吉西他滨的代谢物将被配制在LCP纳米颗粒中，并单独或与siRNA一起递送至肿瘤细胞。这是一种新的方法，利用了这些纳米颗粒的大负载能力。该项目具有里程碑意义，到今年年底，将确定至少一种纳米颗粒制剂用于GEM的治疗，药代动力学和毒性研究。该制剂应准备好与NCI的纳米技术表征实验室合作进行详细的表征和毒性研究， 进一步翻译发展