Targeted nanotherapy for the prevention of post-traumatic osteoarthritis

摘要 最新的人口普查表明，美国有5000多万成年人患有某种形式的关节炎，到2030年， 据估计，年龄> 18岁的6700万美国人被诊断患有关节炎。关节炎 (OA)是最常见的关节炎形式，并且是老年人发病的主要原因 但是对于这些疾病的医学治疗有限，并且没有疾病改善药物。虽然有效的疾病- 骨关节炎修饰药物（DMOADs）是目前临床急需的药物，但目前尚无一种药物在临床上成功应用。后 创伤性骨关节炎（PTOA）是一种在关节损伤后发生的骨关节炎，其性质和时间 创伤是公知。约12%的总体OA负担可追溯至关节创伤。真 PTOA的患病率可能更高，因为损伤与PTOA之间的延迟时间较长（10-15年）。PTOA 影响了美国560万人。膝关节PTOA的预测终生风险 前交叉韧带（ACL）断裂或半月板损伤的患者占50- 70%。PTOA 占所有关节置换术的20%。退伍军人健康相关性：在服役人群中 成员，PTOA的所有原因导致28%的士兵在某种程度上永久残疾，而 有12%的平民遭受关节创伤。此外，关节炎的频率在战斗老兵从 美国正在进行的军事行动估计占11.8%。与战斗有关的关节炎的诊断 损伤会加速，并且通常伴随着不良结局，需要在较年轻时进行关节置换。 退伍军人人口与普通人口的比例。因此，任何改变曲线和 降低PTOA发展的风险将使普通人群和军队中的患者受益 军人和退伍军人。 研究表明，关节损伤后即刻出现强烈的炎症反应（即 在第一周至2-3个月期间），其持续数月至数年，尽管处于较低水平。这 炎症反应可能导致软骨细胞死亡、软骨修复受损和随后的 以PTOA为特征的软骨退化。我们认为这种早期炎症的调节限制了 急性软骨损伤而递送促进软骨修复/体内平衡的合成代谢因子将 从而进一步延迟或阻止最终PTOA发展。我们采用了一种基于肽的自组装技术 纳米复合物，递送多种核酸[小干扰（si）RNA和信使（m）RNA]和 允许使用相同的细胞穿透肽同时靶向多个途径，而不需要 以用于主干或端件改变。我们显示了我们肽基的关节内（IA）递送 纳米颗粒敲低NF κ B表达可减轻早期建立的PTOA，使NF κ B失稳。 手术诱导PTOA的内侧半月板（DMM）模型。我们还证明了 NF κ B siRNA和过表达WNT 16的mRNA，WNT 16是一种对维持细胞凋亡有重要作用的信号通路。 软骨内环境平衡，进一步减轻了DMM模型中早期建立的PTOA。探索独一无二的 我们提出的自组装肽-核酸纳米平台的潜力，以确定长期结果 在DMM模型中联合抗炎和合成代谢纳米疗法的研究。我们相信这些努力将可能 产生了一种用于PTOA疾病改善纳米治疗方法。从这些研究中获得的见解可能 一般适用于OA治疗。

ABSTRACT The latest census indicates that 50+ million adults in the US bear some form of arthritis and by 2030, an estimated 67 million Americans ages > 18 years are projected to have the diagnosis of arthritis. Osteoarthritis (OA) represents the most common form of arthritis and is a major cause of morbidity in the aging population but for which there are limited medical treatments and no disease modifying drugs. Although effective disease- modifying OA drugs (DMOADs) are critically needed, none has successfully emerged in the clinic. Post- traumatic OA (PTOA) is a form of OA that develops after a joint injury and in which the nature and time of trauma is generally known. Approximately 12% of the overall OA burden can be traced to joint trauma. True prevalence for PTOA may be higher, given the long delay between injury and PTOA (10-15 years). PTOA affects 5.6 million individuals in the US. The projected lifetime risk of developing PTOA of the knee in individuals who sustain anterior cruciate ligament (ACL) rupture or meniscal damage is 50-70%. PTOA accounts for 20% of all joint replacements. Veteran Health Relevance: Among the population of service members, all causes of PTOA result in some degree of permanent disability in 28% of soldiers compared to 12% of civilians who sustain joint trauma. In addition, the frequency of arthritis among combat veterans from ongoing US military operations is estimated at 11.8%. The diagnosis of arthritis resulting from combat-related injury is accelerated and often accompanied by poor outcomes, requiring joint replacement at a younger age in the veteran population compared to the general population. Thus, any intervention that shifts the curve and decreases the risk of PTOA development will benefit the patients in the general population as well as military service members and veterans. Studies have documented a robust inflammatory response in the immediate aftermath of joint injury (i.e. during the first week up to 2-3 months) that persists for months to years, albeit at a lower level. This inflammatory response likely contributes to chondrocyte death, impaired cartilage repair and the subsequent cartilage degeneration that characterizes PTOA. We posit that modulation of this early inflammation limits acute cartilage damage while delivery of an anabolic factor that promotes cartilage repair/homeostasis will further delay or prevent eventual PTOA development. We employed a peptide-based self-assembling nanocomplex, delivering a wide range of nucleic acids [small interfering (si)RNA and messenger (m)RNA] and allowing for the simultaneous targeting of multiple pathways using the same cell-penetrating peptide, with no need for backbone or end-piece alterations. We show that intra-articular (IA) delivery of our peptide-based nanoparticle to knock down expression of NFKB mitigates early established PTOA in the Destabilization of the Medial Meniscus (DMM) model of surgery-induced PTOA. We also demonstrate that simultaneous delivery of NFKB siRNA and mRNA that overexpresses WNT16, a signaling pathway important for the maintenance of cartilage homeostasis, further mitigates early established PTOA in the DMM model. To explore the unique potential of our self-assembling peptide-nucleic nanoplatform we propose to determine the long-term outcomes of combined anti-inflammatory and anabolic nanotherapy in the DMM model. We believe these efforts will likely produce a disease-modifying nanotherapeutic approach for PTOA. Insight gained from these studies may be applicable to OA treatment in general.