A microphysiological system of tendon inflammation and fibrosis for drug screening and efficacy testing

用于药物筛选和疗效测试的肌腱炎症和纤维化的微生理系统

• 批准号: 10674534
• 负责人: Hani A Awad
• 金额: $ 72.19万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-15 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10674534
• 关键词: 3-Dimensional; Acceleration; Acute; Adhesions; Antibodies; Biological; Biological Assay; Biological Markers; Biological Models; Blood Tests; Blood Vessels; Blood flow; Cell Cycle Regulation; Cells; Chronic; Cicatrix; Circulation; Clinical; Clinical Trials; Collagen; Complex; Confocal Microscopy; Connective and Soft Tissue; Detection; Devices; Disease; Disease model; Dose; Drug Screening; Endothelial Cells; Endothelium; Engineering; Enzyme-Linked Immunosorbent Assay; Evaluation; Extracellular Matrix; Extravasation; FDA approved; FRAP1 gene; Fibroblasts; Fibrosis; Flexor; Gamma-H2AX; Gene set enrichment analysis; Goals; Health; Human; Hydrogels; Image; Immunology; Impairment; In Situ; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Intervention Trial; Label; Libraries; Link; Macrophage; Measurement; Measures; Mechanics; Mediating; Membrane; Microfabrication; Microfluidic Microchips; Microfluidics; Microscopic; Microscopy; Molecular; Mus; Musculoskeletal; Myofibroblast; Nanoporous; Operative Surgical Procedures; Optics; Outcome; PI3K/AKT; Paracrine Communication; Pathology; Patients; Periodicity; Permeability; Pharmaceutical Preparations; Phase; Phenotype; Porosity; Procedures; Process; Protein Secretion; Publishing; Replacement Arthroplasty; Role; SDZ RAD; Safety; Sampling; Sensitivity and Specificity; Serum; Side; Signal Transduction; Silicon; Sirolimus; Spinal Fusion; System; Tendinopathy; Tendon Injuries; Tendon structure; Testing; Therapeutic; Time; Tissue Engineering; Tissues; Transforming Growth Factor beta; Translating; Vascular Endothelial Cell; biological development; biomarker selection; cell motility; clinically relevant; design; drug candidate; drug discovery; drug efficacy; druggable target; efficacy evaluation; efficacy testing; experience; healing; high throughput screening; human tissue; improved; induced pluripotent stem cell; induced pluripotent stem cell technology; inhibitor; injured; innovation; intercellular communication; joint function; mTOR Inhibitor; manufacture; meter; microphysiology system; microscopic imaging; mouse model; nano; nanofabrication; next generation sequencing; organ on a chip; photonics; preclinical trial; primary outcome; programs; regenerative therapy; repaired; secondary outcome; senescence; sensor; sensor technology; virtual clinical trial

Acute and chronic tendon injuries are among the most common musculoskeletal health problems. Typically, an injured tendon experiences fibrotic scarring that leaves the tissue mechanically compromised and prone to debilitating adhesions that impair joint function. In a fibrotic tendon scar, the cell-cell and paracrine signaling between inflammatory cells, such as macrophages, and tendon fibroblasts activate the latter into fibroproliferative myofibroblasts, ultimately differentiating into a senescent phenotype. Our previous studies using next-generation sequencing and gene set enrichment analysis mechanistically linked fibrosis and senescence in injured mouse tendons with TGF-beta activated mTOR signaling. To further elucidate this pathology, the goal of this proposal is to engineer a microfluidic human tendon-on-chip (hToC) system and use it to more accurately model the biological aspects of the inflammation and fibrosis in injured tendons. In the UG3 phase of this proposal, the chip will be fabricated featuring a multicompartmental design and microfluidic channels to incorporate a fibroblast-seeded collagen hydrogel and simulate vascular blood flow, respectively. Ultrathin, highly permeable, and optically transparent porous silicon membranes (SiM) will separate the hydrogel from circulation and provide a substrate for an endothelial barrier in between. The signaling between the fibroblasts, hydrogel-resident- and circulating-macrophages, and endothelial cells will be enabled through nanoporous SiM (~60 nm), while a microporous SiM (~ 8 µm) will allow extravasation of circulating macrophages and infiltration of the hydrogel under TGF-beta stimulation. To allow for a patient-centric chip, tendon fibroblasts will be used to create the tendon hydrogel and to reprogram donor-matching iPSCs to derive the endothelial cells and macrophages, respectively. An additional innovation will be the integration of label- free photonic sensors into the microfluidic device to allow on-chip sensing, which has been long appreciated as a critical, unmet need for organ-on-chip devices. The UG3 studies will use the chip to validate the role of mTOR in the disease model and identify biologically relevant biomarkers. In the UH3 phase, we will utilize the chip as a pre-clinical trial platform for testing efficacy and safety of FDA-approved mTOR inhibitors as potential disease modifying drugs, and as a drug screening platform to identify and prioritize safer and more potent inhibitors of mTOR and senescence in tendon injury for clinical trials. To successfully complete this innovative project, we have assembled a team of accomplished experts in tendon tissue engineering and surgery, immunology, iPSC technology, GMP cell manufacturing, nano- and micro-fabrication, sensor technology, and high throughput screening. The proposed studies will develop a human microphysiological system to catalyze clinical trials and accelerate drug discovery for acute and chronic tendon injuries.

急性和慢性肌腱损伤是最常见的肌肉骨骼健康问题之一。通常，一个 受伤的肌腱会经历纤维性疤痕，使组织机械受损，并容易 损害关节功能的衰弱粘连。在纤维性肌腱瘢痕中，细胞-细胞和旁分泌信号 在炎症细胞，如巨噬细胞和肌腱成纤维细胞之间，激活后者进入 纤维增生性肌成纤维细胞，最终分化为衰老表型。我们之前的研究 使用下一代测序和基因集浓缩分析机制相关的纤维化和 转化生长因子-β激活的mTOR信号在损伤的小鼠肌腱中的衰老。为了进一步阐明这一点 病理学，这项提议的目标是设计一种微流控人体肌腱芯片(HToC)系统并使用 它可以更准确地模拟损伤肌腱的炎症和纤维化的生物学方面。在 这项建议的UG3阶段，将制造的芯片具有多分区设计和微流控 分别加入成纤维细胞种植的胶原水凝胶和模拟血管血流的通道。 超薄、高渗透性和光学透明的多孔硅膜(SIM)将把 水凝胶从循环中释放，并为介于两者之间的内皮屏障提供底物。之间的信令 成纤维细胞、水凝胶驻留和循环中的巨噬细胞以及内皮细胞将通过 纳米孔SIM(~60 nm)，而微孔SIM(~8微米)将允许循环外渗 在转化生长因子-β刺激下，巨噬细胞和水凝胶的渗透。为了实现以患者为中心的芯片， 肌腱成纤维细胞将被用来产生肌腱水凝胶，并对供体匹配的IPSCs进行重新编程，以获得 分别为内皮细胞和巨噬细胞。另一项创新将是Label- 在微流控器件中加入自由光子传感器以实现片上传感，这一直被认为是 对芯片上器官设备的关键的、未得到满足的需求。UG3的研究将使用该芯片来验证 MTOR在疾病模型中的作用，并确定与生物相关的生物标志物。在UH3阶段，我们将利用 芯片作为临床前试验平台测试FDA批准的mTOR抑制剂的有效性和安全性 疾病修饰药物，并作为药物筛选平台，识别和优先考虑更安全和更有效的药物 肌腱损伤中mTOR和衰老的抑制剂进行临床试验。要成功完成这一创新 项目，我们已经组建了一支在肌腱组织工程和外科手术方面经验丰富的专家团队， 免疫学，IPSC技术，GMP细胞制造，纳米和微型制造，传感器技术，以及 高通量筛选。拟议中的研究将开发一种人体微生理系统来催化 临床试验和加速急、慢性肌腱损伤的药物发现。