Topical Delivery of siRNA Nanconjugates: Suppressing Epidermal Hyperplasia

siRNA 纳米缀合物的局部递送：抑制表皮增生

基本信息

批准号：
8632993
负责人：
CHAD A. MIRKIN
金额：
$ 33.53万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-03-01 至 2016-02-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8632993
关键词：
Address Affinity Binding Biology Cancer Cell Growth Cell physiology Cells Characteristics Charge Clinical Communities Coupled Dominant-Negative Mutation Down-Regulation Endocytosis Pathway Engineering Epidermal Growth Factor Receptor Epidermis Foundations Gene Delivery Gene Expression Gene Silencing Gene Targeting Genes Goals Gold Green Fluorescent Proteins HRAS gene Hereditary Disease Histologic Human Hyperplasia Immunoblotting In Vitro Inflammation Investigation Laboratories Laboratory mice Lead Mass Spectrum Analysis Membrane Methods Modality Modeling Mus Mutation Nanoconjugate Nanotechnology Nude Mice Oligonucleotides Pathologic Penetration Pharmaceutical Preparations Phase I Clinical Trials Phenotype Plasma Point Mutation RNA Research Safety Signal Transduction Skin Skin Tissue Small Interfering RNA Specificity Stratum corneum Surface System Testing Topical application Toxic effect Transmission Electron Microscopy Transplantation Viscera Work clinical application clinical practice density gene function gene therapy human disease in vivo inhibitor/antagonist intravenous administration keratinocyte knock-down mouse model mutant nanoparticle novel nuclease overexpression pre-clinical skin disorder small molecule tool uptake

项目摘要

DESCRIPTION (provided by applicant): Short interfering ribonucleic acids (siRNAs) are proven research tools that have revolutionized our ability to suppress genes. However, delivery remains a major hurdle for their use in vivo, including the limited ability of siRNAs to traverse the highly effective epidermal barrier after topical application. The objective of this project is to optimize and evaluate a topically applied nanoparticle delivery system for siRNA that is able to penetrate the skin and selectively suppress gene expression. Such a delivery system would have the potential to selectively target even small mutations that lead to genetic disorders, with implications ranging from inhibiting cancer cell growth to suppressing inflammation. Our laboratories have engineered a novel nanoparticle conjugate utilizing siRNA duplexes that are densely packed on the surface of gold nanoparticles (siRNA-Au NPs) and which demonstrates a surprising ability to transit the mouse and human stratum corneum and suppress two tested targets, green fluorescent protein and the epidermal growth factor receptor. To date, no toxicity has been found when the siRNA-Au NPs are delivered either topically or intravenously into mice at concentrations that far exceed those needed to suppress genes. In our proposed work, we will determine the mechanism(s) by which siRNA-Au NPs penetrate into human skin, and use this information to optimize siRNA-Au NPs for gene suppression. Next, we will assess the ability of siRNA-Au NPs to suppress Ras signaling and reverse epidermal hyperplasia in a mouse model of skin-specific, inducible overexpression of H-Ras. Building on our in vitro and in vivo mouse studies with siRNA-Au NPs, we will test if topical application of siRNA-Au NPs to a human transplanted skin model of Ras overexpression will similarly suppress aberrant Ras signaling to cause clinically and histologically detectable epidermal normalization. This work will lay the foundation for clinical application of the siRNA nanoparticle conjugates, and establish a new paradigm for the topical application of gene therapies.

描述（由申请人提供）：短干扰核糖核酸 (siRNA) 是经过验证的研究工具，彻底改变了我们抑制基因的能力。然而，递送仍然是它们在体内使用的主要障碍，包括局部应用后 siRNA 穿过高效表皮屏障的能力有限。该项目的目标是优化和评估局部应用的 siRNA 纳米颗粒递送系统，该系统能够穿透皮肤并选择性抑制基因表达。这种递送系统有可能选择性地靶向导致遗传性疾病的微小突变，其影响范围从抑制癌细胞生长到抑制炎症。我们的实验室利用密集堆积在金纳米颗粒 (siRNA-Au NP) 表面的 siRNA 双链体设计了一种新型纳米颗粒缀合物，该缀合物表现出令人惊讶的穿过小鼠和人类角质层并抑制两个测试目标（绿色荧光蛋白）的能力和表皮生长因子受体。迄今为止，当 siRNA-Au NP 以远远超过抑制基因所需的浓度局部或静脉注射到小鼠体内时，尚未发现任何毒性。在我们提出的工作中，我们将确定 siRNA-Au NP 渗入人体皮肤的机制，并利用这些信息来优化 siRNA-Au NP 进行基因抑制。接下来，我们将在皮肤特异性、诱导性 H-Ras 过度表达的小鼠模型中评估 siRNA-Au NP 抑制 Ras 信号传导和逆转表皮增生的能力。基于我们对 siRNA-Au NP 进行的体外和体内小鼠研究，我们将测试将 siRNA-Au NP 局部应用于 Ras 过度表达的人类移植皮肤模型是否会同样抑制异常 Ras 信号传导，从而导致临床和组织学可检测到的表皮正常化。这项工作将为siRNA纳米颗粒缀合物的临床应用奠定基础，并为基因治疗的局部应用建立新的范例。