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（由申请人提供）：NF-κ B信号通路代表慢性炎症的有吸引力的治疗靶点，因为它调节许多控制炎症反应的基因产物。然而，这种途径的普遍抑制可导致严重的宿主毒性，因为NF-κ B是先天性和适应性免疫两者的主要介质。我们已经制定了一种新的肽-siRNA构建体，其特征在于，并评估了其自凝聚，在血清中稳定，并基于两亲性阳离子肽蜂毒肽（蜂毒的一种组分，其通常通过寡聚化在膜中形成孔）的修饰版本，以内在的内体释放机制将寡核苷酸递送至细胞质。我们发现，修饰的蜂毒肽可以缀合到无数的货物和运输材料迅速进入细胞质，有效的基因沉默： 靶向NF-κ B p65典型亚基的复合物通过渗漏的脉管系统归巢到发炎的爪子，并有效地抑制了RA小鼠模型中正在进行的炎症。广泛的体内和离体研究还表明，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。在本提案中，我们将使用肽-siRNA平台来探索双重方法（靶向NF-κ B经典和非经典途径）在治疗RA中具有协同或累加益处的假设。为此，我们将配制和优化含有NF-κ B-p65（规范）、p52/100（非规范）或两者组合的肽- siRNA构建体，并在人/鼠细胞系中体外和在两种炎性关节炎鼠模型中体内测试它们对NF-κ B信号传导途径敲低的效率。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，其他疾病的过程，是服从RNA干扰治疗。