In situ Cell Engineering for On-demand TIMP Expression in Osteoarthritis

用于骨关节炎按需表达 TIMP 的原位细胞工程

• 批准号: 10451707
• 负责人: Jonathan Matthew Brunger
• 金额: $ 15.69万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-15 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10451707
• 关键词: Address; Adrenal Cortex Hormones; Aging; Animals; Behavior; Biochemical; Biological; Biology; Cartilage; Cells; Chemistry; Chondrocytes; Chronic; Chronic Disease; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Collagen; Data; Degenerative polyarthritis; Deterioration; Diagnosis; Disease; Disease Progression; Divalent Cations; Dose; Drug Targeting; Economic Burden; Endosomes; Equilibrium; Feedback; Formulation; Gene Targeting; Genes; Genome; Genome engineering; Goals; Guide RNA; Hybrids; In Situ; In Vitro; Incidence; Inflammation; Inflammatory; Injections; Injury; Interleukin-1; International; Intervention; Intra-Articular Injections; Joints; Kinetics; Knee; Knock-in; Knock-out; Length; Life; Light; Longevity; Malignant Neoplasms; Matrix Metalloproteinase Inhibitor; Matrix Metalloproteinases; Measures; Mechanics; Mediating; Modeling; Mus; Non-Steroidal Anti-Inflammatory Agents; Outcome; Pain; Pathologic; Pathology; Patients; Pharmaceutical Preparations; Pharmacology; Pharmacotherapy; Phenotype; Polymer Chemistry; Polymers; Prevalence; Production; Proteins; Quality of life; RNA Interference; Replacement Arthroplasty; Reporter; Research; Resistance; Safety; Signal Transduction; Silicon; Single-Stranded DNA; Societies; Steroids; Stimulus; Stomach; Structural Protein; Switch Genes; System; TIMP3 gene; Technology; Testing; Therapeutic; Thinness; Tissue Inhibitor of Metalloproteinases; Tissues; Toxic effect; Transgenes; Treatment Efficacy; aggrecanase; angiogenesis; arm; arthropathies; base; cartilage degradation; cellular engineering; collagenase 3; cytokine; cytotoxicity; disability; efficacious treatment; experience; fluorescence imaging; gene therapy; genome editing; histological image; improved; in vivo; in vivo imaging system; inhibitor; innovation; insight; joint destruction; joint injury; mechanical load; nanoparticle; nucleic acid-based therapeutics; palliation; palliative; promoter; prophylactic; red fluorescent protein; side effect; small molecule; success; synthetic biology; therapeutic target; transgene expression

Osteoarthritis (OA) is a prevalent degenerative joint disorder and the leading cause of disability. Presently, there are no disease-modifying osteoarthritis drugs (DMOADs). Diagnosis and pharmacological intervention occur mostly at a late stage, and current treatments offer only temporary, palliative relief before disease progression necessitates joint replacement. OA prevalence is high, at roughly 27% of those over 40 years old, and occurrence of post-traumatic OA (PTOA) is even higher (over 50%) following injury of large joints such as the knee. Given its high incidence and predictability, PTOA has potential to be treated prophylactically, a strategy that is both conducive to achieving disease-modifying outcomes and commercially/clinically feasible, provided treatment offers long-term protection. We aim to achieve persistent joint protection by permanently converting cells in situ into “on-demand” TIMP-3 “factories”, harnessing TIMP-3 as a pan-MMP inhibitor that blocks multiple aspects of OA pathology, including cartilage degradation, angiogenesis, and inflammation. “On-demand” expression of TIMP-3 will be achieved via a targeted and permanent gene insertion that hijacks the Mmp13 promoter. This approach is based on a nonviral CRISPR-based nanoparticle and activates TIMP-3 expression only when pathological (OA) stimuli are present, minimizing potential side-effects. We propose to optimize a nanoparticle formulation for non-viral gene knock-in and quantify the therapeutic efficacy of TIMP-3 knock-in in vitro and in vivo. This therapy has potential to avoid significant loss in quality of life for patients who experience a large joint injury and is uniquely enabled by our team with expertise in intracellular delivery (Duvall), polymer and nanoparticle chemistry (D’Arcy), genome editing and synthetic biology (Brunger), and PTOA biology (Hasty).

骨关节炎是一种常见的退行性关节疾病，也是致残的主要原因。目前，在那里 没有治疗骨关节炎的药物(DMOADs)。诊断和药物干预 主要是在晚期，目前的治疗方法只能在疾病进展之前提供暂时的姑息缓解 有必要进行关节置换。骨性关节炎的患病率很高，在40岁以上的人群中约占27%， 在诸如膝盖等大关节损伤后，创伤后骨性关节炎(PTOA)的发生率更高(超过50%)。vt.给出 由于其高发病率和可预测性，PTOA有可能被预防性治疗，这一战略既是 有利于实现改善疾病的结果，并在商业/临床上可行，提供治疗 提供长期保护。我们的目标是通过在原位永久转换细胞来实现持久的联合保护 将TIMP-3作为一种泛-基质金属蛋白酶抑制剂，阻断多个方面的 骨关节炎病理，包括软骨退化、血管生成和炎症。“按需”的表达方式 TIMP-3将通过劫持Mmp13启动子的靶向和永久性基因插入来实现。这 该方法基于基于CRISPR的非病毒纳米颗粒，仅在以下情况下激活TIMP-3表达 存在病理性(OA)刺激，将潜在的副作用降至最低。我们建议优化一种纳米颗粒 非病毒基因敲入模型的建立及TIMP-3体外和体内治疗效果的定量研究 活着。这种疗法有可能避免经历大关节的患者的生活质量显著下降。 我们的团队在细胞内递送(Duvall)、聚合物和 纳米粒子化学(D‘Arcy)、基因组编辑和合成生物学(Brunger)和PTOA生物学(Hats)。