Extracellular vesicles as therapeutic vehicles for chondroprotection

细胞外囊泡作为软骨保护的治疗载体

• 批准号: 10268960
• 负责人: Ryan Michael Porter
• 金额: $ 30.06万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-16 至 2023-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10268960
• 关键词: Acute; Animal Model; Cartilage; Cartilage Matrix; Cells; Charge; Chondrocytes; Collagen; Degenerative polyarthritis; Development; Diffusion; Electrostatics; Genes; Glycosaminoglycans; Homeostasis; In Vitro; Inflammatory; Injury; Mediating; Mesenchymal; Metabolism; Modeling; Musculoskeletal Diseases; Operative Surgical Procedures; Paracrine Communication; Penetration; Prevention; Research; Signal Transduction; Stromal Cells; Structural defect; Surface; Synovial joint; Testing; Therapeutic; Tissues; Vesicle; articular cartilage; critical period; design; extracellular vesicles; gain of function; genome-wide; improved; joint function; joint injury; paracrine; preclinical study; prevent; regenerative; solute; stem; uptake

A single joint injury can result in the development of post-traumatic osteoarthritis (PTOA). Central to loss of joint function is the progressive degeneration of articular cartilage. While surgical procedures can correct structural defects, there are no treatments that can protect chondrocyte metabolism and viability in the weeks after an injury. Preclinical studies have shown that mesenchymal stem/stromal cells (MSCs) grown in vitro can have chondroprotective effects in animal models of PTOA; these effects are now thought to be mediated by MSC paracrine signaling following intra-articular delivery. Recently, MSC-derived extracellular vesicles (MSC-EVs) have been shown to stimulate pro-regenerative effects equivalent to their donor cells in several non-skeletal injury models, suggesting they are an important component of MSC paracrine actions. The potential of MSC-EVs for preventing or delaying PTOA may depend on their ability to deliver signals directly to chondrocytes, which requires transport through the dense, negatively-charged matrix of cartilage. While EV size and net charge suggest limited penetration within healthy cartilage, their precise surface composition may influence cartilage uptake and diffusion. Moreover, cartilage matrix changes that occur during the acute inflammatory period after a joint injury can alter the transport of larger solutes, and EVs may respond similarly. If MSC-EVs can deliver signals that improve chondrocyte function before irreversible changes occur to the cartilage matrix, they can be used to restore tissue homeostasis during the critical period following joint trauma. This project will better define the therapeutic potential of MSC-EVs for PTOA through an improved understanding of the factors regulating their cargo delivery to chondrocytes. We will first test whether cartilage glycosaminoglycan depletion, a more reversible change to the cartilage matrix than collagen loss, impacts MSC-EV cargo delivery (Aim 1). We will also determine how delivery is influenced by MSC-EV surface charge and altered electrostatic interactions with the cartilage matrix (Aim 2). Finally, we will identify chondrocyte genes that control MSC-EV cargo delivery through a genome-wide gain-of-function screen (Aim 3). In the short term, this project will lay the groundwork for studying potential EV modes of action for the prevention of PTOA; in the long term, it will inform the design of synthetic vesicles for delivering therapeutic molecules to synovial joint cells.

单个关节损伤可导致创伤后骨关节炎(PTOA)的发展。关节功能丧失的核心是关节软骨的进行性退化。虽然外科手术可以纠正结构缺陷，但没有治疗方法可以在受伤后的几周内保护软骨细胞的新陈代谢和生存能力。临床前研究表明，在体外培养的间充质干细胞(MSCs)在PTOA动物模型中可以起到软骨保护作用；目前认为这些作用是由MSC关节内释放后的旁分泌信号介导的。最近，在几种非骨骼损伤模型中，MSC来源的细胞外小泡(MSC-EV)被证明具有与供体细胞相同的促再生作用，这表明它们是MSC旁分泌作用的重要组成部分。MSC-EVS预防或延迟PTOA的潜力可能取决于它们直接向软骨细胞传递信号的能力，这需要通过致密的负电荷软骨基质运输。虽然EV的大小和净电荷表明其在健康软骨中的穿透性有限，但其精确的表面组成可能会影响软骨的摄取和扩散。此外，关节损伤后急性炎症期发生的软骨基质变化可以改变较大溶质的运输，EVS也可能做出类似的反应。如果MSC-EVS能够在软骨基质发生不可逆转的变化之前发出改善软骨细胞功能的信号，它们可以用于在关节创伤后的关键时期恢复组织内稳。该项目将通过更好地了解调节MSC-EVS向软骨细胞运送货物的因素，更好地确定MSC-EVS治疗PTOA的潜力。我们将首先测试软骨糖胺多聚糖的耗竭是否会影响MSC-EV的货物运输(目标1)。糖胺聚糖耗竭是软骨基质比胶原蛋白丢失更可逆的变化。我们还将确定MSC-EV表面电荷和改变的静电与软骨基质的相互作用是如何影响释放的(目标2)。最后，我们将通过全基因组的功能获得筛选来确定控制MSC-EV货物运输的软骨细胞基因(目标3)。在短期内，该项目将为研究预防PTOA的潜在EV作用模式奠定基础；从长期来看，它将为设计用于向滑膜关节细胞输送治疗分子的合成囊泡提供信息。