Inhibition of NF-kB with siRNA/Peptide Nanoparticles for Arthritis Treatment

用 siRNA/肽纳米颗粒抑制 NF-kB 治疗关节炎

• 批准号: 9025681
• 负责人: Christine T. Pham
• 金额: $ 36.27万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9025681
• 关键词: Adverse effects; Affect; Antibody Formation; Antiviral Response; Arthritis; Bee Venoms; Biodistribution; Blood Vessels; Cartilage; Cell Line; Cells; Chronic; Complement Activation; Complex; Connective Tissue; Cytoplasm; Data; Development; Disease; Dose; Drug Kinetics; Drug or chemical Tissue Distribution; Endothelial Cells; Enzymes; Fibroblasts; Formulation; Gene Silencing; General Population; Goals; Health; Home environment; Human; Immune response; Immune system; In Vitro; Inflammation; Inflammatory; Inflammatory Arthritis; Inflammatory Response; Interferon Type I; Joints; Lead; Ligands; Lymphocyte; Mediating; Mediator of activation protein; Medical; Membrane; Morbidity - disease rate; Mus; NF-kappa B; Natural Immunity; Oligonucleotides; Pathway interactions; Patients; Peptides; Phase; Process; RNA Interference Therapy; Regulatory T-Lymphocyte; Rheumatoid Arthritis; Serum; Signal Pathway; Small Interfering RNA; Specificity; Structure; Systems Development; T-Lymphocyte; Testing; Therapeutic; Therapeutic Agents; Toxic effect; adaptive immunity; arthropathies; base; bone; collagen antibody induced arthritis; cytokine; design; disability; gene product; immunogenicity; in vitro testing; in vivo; joint destruction; knock-down; macrophage; monocyte; mouse model; nanomedicine; nanoparticle; novel; novel therapeutics; p65; response; success; therapeutic target

 DESCRIPTION (provided by applicant): The NF-KB signaling pathway represents an attractive therapeutic target for chronic inflammation as it regulates many gene products that control the inflammatory response. However, generalized suppression of this pathway can result in serious host toxicity, as NF-KB is a major mediator of both innate and adaptive immunity. We have formulated, characterized, and evaluated a novel peptide-siRNA construct that is self-condensing, stable in serum, and delivers oligonucleotides to the cell cytoplasm with an intrinsic endosomal releasing mechanism based on a modified version of the amphipathic cationic peptide melittin, a component of bee venom that normally forms pores in membrane by oligomerization. We showed that the modified melittin could be conjugated to myriad cargos and transport materials rapidly into the cell cytoplasm for efficient gene silencing: The peptide-siRNA complexes targeting the NF-KB p65 canonical subunit homed to the inflamed paws through leaky vasculature and potently suppressed ongoing inflammation in a murine model of RA. Extensive in vivo and ex vivo studies also suggest that the platform has a favorable toxicity profile and did not elicit immunogenicity after repeated dosing. In this proposal we will use the peptide-siRNA platform to explore the hypothesis that a dual approach (targeting both NF-KB canonical and non-canonical pathways) will have synergistic or additive benefits in the treatment of RA. To that end we will formulate and optimize peptide- siRNA constructs containing NF-KB-p65 (canonical), p52/100 (non-canonical), or a combination of both and test their efficiency for NF-KB signaling pathway knockdown in vitro in human/murine cell lines and in vivo in two murine models of inflammatory arthritis. The results will provide in vivo proof-of-concept for this new peptide- siRNA platform and the success will justify further development of this system for other disease processes that are amenable to RNA interference therapy.

 描述（由适用提供）：NF-KB信号通路代表了慢性感染的有吸引力的治疗靶标，因为它调节了许多控制炎症反应的基因产物。但是，由于NF-KB是先天和适应性免疫组织化学的主要介体，因此对该途径的普遍抑制可能导致严重的宿主毒性。 We have formulated, characterized, and evaluated a novel peptide-siRNA construct that is Self-condensing, stable in serum, and deliver oligonucleotides to the cell cytoplasm with an intrinsic endosomal release mechanism based on a modified version of the amphipathic cationic peptide melittin, a component of bee venom that normally forms pores in membrane by oligomerization.我们表明，修饰的蜂素可以偶联到无数的嘉戈斯，并迅速将材料转运到细胞质中以进行有效的基因沉默：肽 - siRNA 针对NF-KB P65典型亚基的复合物通过漏水的脉管系统归为发炎的爪子，并可能在RA的鼠模型中抑制了持续的感染。大量体内和体内研究还表明，该平台具有良好的毒性特征，并且在重复给药后不会引起免疫原性。在此提案中，我们将使用胡椒 - siRNA平台来探讨以下假设：双重方法（针对NF-KB规范和非典型途径）将在RA治疗中具有协同或上瘾的益处。为此，我们将制定和优化含有NF-KB-P65（规范），p52/100（非典型）的肽siRNA构建体，或者两者结合并测试其NF-KB信号通路在人/鼠类/鼠类鼠类/鼠类鼠类/鼠类鼠的炎症模型中的体外敲低的效率。结果将为此提供体内概念验证 新的肽siRNA平台和成功将证明该系统的进一步发展是针对RNA干扰疗法的其他疾病过程的。