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，例如与运动有关的关节损伤。这是关节的常见原因。 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。作为诺贝尔奖获得者的发现，小干扰RNA (SiRNA)提供了很大的治疗潜力，可以特异性地抑制疾病基因的表达。然而，它是 传递带负电荷的siRNA以穿透无血管、致密、带负电荷的siRNA具有极大的挑战性 软骨基质。此外，关节胶囊中的治疗药物通常被迅速清除，限制了它们的驻留。 时间短至1-5个小时。为了克服这些障碍，我们开发了一种非共价Janus基纳米 名为Nanopiess(NP)的送货工具，具有定制的尺寸和表面属性，使 包裹的siRNA可穿透关节软骨组织并进入软骨细胞。通过与…绑定 细胞外基质(ECM)，NP在软骨中保留较长的半衰期，将屏障转化为载体。这个 这项提案的目标是确定调节NPs尺寸和表面性质的因素， 从而1)确定有效地渗透到软骨中的NPs的最佳尺寸；2)制定 结合软骨基质以用于组织保持的纳米颗粒的表面性质，以及3)治疗能力的评估 抑制DMM模型PTOA进展的NPs。基本的理论基础是 这一提议将1)促进我们对非共价纳米递送车辆自组装的理解 以及它们与组织ECM分子的相互作用；2)实现了一种平台siRNA递送技术， 穿透软骨和其他富含基质的组织，具有定制的尺寸和表面特性；以及3) 为研制首个针对PTOA的疾病修饰RNA疗法奠定了基础。这个 本文的创新之处在于：1)NP是一种新一代药物载体，具有独特的性能 具有尺寸通用性、对细胞外基质分子的亲和力和良好的生物降解性等优点 无毒。2)我们描述了输送载体与软骨细胞外基质之间的相互作用 载体的尺寸和表面性质，进而决定软骨内植入成功的关键因素 送货。3)这一技术突破为治疗PTOA提供了一种携带siRNA的治疗方法。 在完成了具体目标后，我们希望1)提高对非 共价纳米材料自组装及其与组织基质的相互作用，2)实现高效和 3)通过siRNA/NP抑制DMM模型的PTOA进展 心理治疗。这些成果将对开发特定的药物输送载体产生重要的积极影响 对于软骨或其他富含基质的组织，以及改善关节疾病的治疗，如PTOA。

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.