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

基本信息

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
项目类别：
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

项目摘要

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（OA）是一种普遍存在的退行性关节疾病，也是目前导致残疾的主要原因。没有发生改变疾病的骨关节炎药物（DMOAD）。大多数处于晚期阶段，目前的治疗只能在疾病进展之前提供暂时的姑息缓解 OA 患病率很高，40 岁以上人群中约 27%，且发生率较高膝关节等大关节受伤后，创伤后 OA (PTOA) 的发生率甚至更高（超过 50%）。由于其高发病率和可预测性，PTOA 有可能进行预防性治疗，这是一种既可有利于实现疾病缓解结果并且商业/临床可行，提供治疗我们的目标是通过原位永久转化细胞来实现持久的关节保护。进入“按需”TIMP-3“工厂”，利用 TIMP-3 作为泛 MMP 抑制剂，阻断多个方面 OA 病理学，包括软骨退化、血管生成和炎症的“按需”表达。 TIMP-3 将通过劫持 Mmp13 启动子的有针对性的永久基因插入来实现。该方法基于非病毒 CRISPR 纳米颗粒，仅在以下情况下激活 TIMP-3 表达：存在病理（OA）刺激，最大限度地减少潜在的副作用我们建议优化纳米颗粒。用于非病毒基因敲入的制剂并量化 TIMP-3 敲入的体外和体内治疗效果这种疗法有可能避免大关节患者生活质量的显着下降。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) 以及 PTOA 生物学 (Hasty)。