Developing Nanomaterial Platform for Intra-Cartilage Delivery of RNA Therapeutics against Joint Diseases

摘要 PTOA是由身体创伤引起的，如运动相关的关节损伤。这是一个共同的原因， 退化和残疾，每年影响560万美国人。目前，没有疾病- 修改药物以防止PTOA进展。作为一项获得诺贝尔奖的发现，小干扰RNA siRNA提供了特异性抑制疾病基因表达的巨大治疗潜力。但据 递送带负电荷的siRNA以穿透无血管的、致密的、带负电荷的 软骨基质此外，关节囊中的治疗剂通常被迅速清除，限制了它们的驻留 时间短至1-5小时。为了克服这些障碍，我们开发了一种非共价Janus基纳米- 名为Nanopiece（NP）的运载工具，具有定制的尺寸和表面特性， 包封的siRNA渗透到关节软骨组织中并进入软骨细胞。通过与 在细胞外基质（ECM）中，NP保留在软骨中长半衰期，将屏障转化为载体。的 该建议的目的是确定调节纳米颗粒尺寸和表面性质的因素， 从而1）鉴定有效渗透到软骨中的NP的最佳尺寸; 2）配制 结合软骨基质用于组织保留的NP的表面性质，和3）评估治疗能力 抑制DMM模型中PTOA进展。基本原理是， 这一建议将1）推进我们对非共价纳米运载工具自组装的理解 以及它们与组织ECM分子的相互作用; 2）实现平台siRNA递送技术， 以定制的尺寸和表面特性穿透软骨和其他富含基质的组织;以及3） 为开发第一种针对PTOA的疾病修饰RNA治疗剂奠定基础。的 提出的研究是创新的，因为：1）NP是新一代药物递送载体，具有独特的 优点如尺寸的多功能性、对ECM分子的亲和性和优异的生物降解性， 无毒性。2)我们描述了传递载体和软骨ECM之间的相互作用， 车辆的尺寸和表面性能，然后确定成功的关键因素， 交付. 3)该技术突破启发了递送siRNA以治疗PTOA的治疗方法。 在实现具体目标后，我们期望：1）推进对非物质文化遗产的基本认识， 共价纳米材料自组装及其与组织基质的相互作用，2）实现高效， 长效siRNA递送到软骨中，和3）通过siRNA/NP抑制DMM模型中的PTOA进展 疗法这些结果将对开发特定的药物递送载体产生重要的积极影响 用于软骨或其他基质丰富的组织，并改善关节疾病如PTOA的治疗。

Abstract PTOA is caused by physical trauma such as sports related joint injuries. It is a common cause of joint degeneration and disability, affecting 5.6 millions of Americans every year. Currently, there is no disease- modifying drug to prevent PTOA progression. As a Nobel-prize winning discovery, small interfering RNA (siRNA) provides great therapeutic potential to specifically inhibit disease gene expression. However, it is extremely challenging to deliver negatively-charged siRNA to penetrate avascular, dense, negatively-charged cartilage matrix. Moreover, therapeutics in the joint capsule is usually cleared rapidly, limiting their residence times to as short as 1-5 hours. To overcome these obstacles, we developed a non-covalent Janus-base nano- delivery vehicle named Nanopiece (NP) with customized dimensions and surface properties, which enable the encapsulated siRNA to penetrate into articular cartilage tissue and enter chondrocytes. Through binding with extracellular matrix (ECM), NP is retained in cartilage for a long half-life, converting a barrier to carrier. The goal of this proposal is to determine the factors regulating the dimensions and surface properties of NPs, thereby 1) identifying the optimal dimensions of NPs that penetrate into cartilage effectively; 2) formulating surface properties of NPs that bind cartilage matrix for tissue retention, and 3) evaluating the therapeutic ability of the NPs to inhibit PTOA progression in the DMM model. The underlying rationale is that the completion of this proposal will 1) advance our understanding on the self-assembly of non-covalent nano-delivery vehicles and their interactions with tissue ECM molecules; 2) realize a platform siRNA delivery technology that penetrating cartilage and other matrix-rich tissues with customized dimensions and surface properties; and 3) lay the foundation for the development of the first disease-modifying RNA therapeutic against PTOA. The proposed research is innovative because: 1) NP is a new generation drug delivery vehicle with unique advantages such as the versatility in dimensions, affinity to ECM molecules and excellent biodegradability and non-toxicity. 2) We delineate the interactions between delivery vehicles and cartilage ECM in terms of the vehicles’ dimension and surface property, and then determine the key factors for successful intra-cartilage delivery. 3) The technology breakthrough enlightens a therapeutic approach to deliver siRNA to treat PTOA. After accomplishing the specific aims, we expect to 1) advance fundamental understandings of the non- covalent nanomaterial self-assembly and its interactions with tissue matrix, 2) achieve highly effective and long-lasting siRNA delivery into cartilage, and 3) inhibit PTOA progression in the DMM model via the siRNA/NP therapy. These outcomes will have important positive impact on developing specific drug delivery vehicles for cartilage or other matrix-rich tissues, and improving treatment of joint diseases such as PTOA.