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

基本信息

批准号：
10239102
负责人：
Hani A Awad
金额：
$ 75.53万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-15 至 2022-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10239102
关键词：
3-Dimensional Acute Adhesions Antibodies Biologic Development Biological Biological Assay Biological Markers Biological Models Blood Circulation Blood Tests Blood Vessels Blood flow Cell Cycle Regulation Cells Chronic Cicatrix Clinical Clinical Trials Collagen Complex Confocal Microscopy Connective and Soft Tissue Crystallization 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 Liquid substance Measurement Measures Mechanics Mediating Membrane 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 Procedures Process Proteins 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 base 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/antagonist injured innovation intercellular communication joint function mTOR Inhibitor macrophage microphysiology system microscopic imaging mouse model nano nanofabrication next generation sequencing organ on a chip photonics preclinical trial primary outcome regenerative therapy repaired secondary outcome senescence sensor sensor technology virtual clinical trial

3-Dimensional Acute Adhesions Antibodies Biologic Development Biological Biological Assay Biological Markers Biological Models Blood Circulation Blood Tests Blood Vessels Blood flow Cell Cycle Regulation Cells Chronic Cicatrix Clinical Clinical Trials Collagen Complex Confocal Microscopy Connective and Soft Tissue Crystallization 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 Liquid substance Measurement Measures Mechanics Mediating Membrane 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 Procedures Process Proteins 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 base 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/antagonist injured innovation intercellular communication joint function mTOR Inhibitor macrophage microphysiology system microscopic imaging mouse model nano nanofabrication next generation sequencing organ on a chip photonics preclinical trial primary outcome regenerative therapy repaired secondary outcome senescence sensor sensor technology virtual clinical trial

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项目摘要

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，一个受伤的肌腱经历纤维化疤痕，使组织机械地妥协并容易损害关节功能的衰弱的依从性。在纤维化肌腱疤痕中，细胞细胞和旁分泌信号传导在炎性细胞（例如巨噬细胞）和肌腱成纤维细胞之间，将后者激活纤维增生性肌纤维细胞，最终区分为感觉表型。我们以前的研究使用下一代测序和基因集富集分析机械上连接的纤维化和带有TGF-β激活的MTOR信号传导的受伤小鼠肌腱的感应。进一步阐明这一点病理学，该提案的目的是设计微流体的人肌腱（HTOC）系统并使用它可以更准确地模拟受伤肌腱注射和纤维化的生物学方面。在该提案的UG3阶段，芯片将由多个剖面设计和微流体制造。分别结合成纤维细胞种子胶原凝胶并分别模拟血管血流的通道。 Ultrathin，高度渗透性和光学透明的多孔硅膜（SIM）将分开从循环中的水凝胶，并为两者之间的内皮屏障提供底物。之间的信号将通过纳米孔SIM（〜60 nm），而微孔SIM（〜8 µm）将允许循环渗出在TGF-β刺激下，巨噬细胞和水凝胶的浸润。为了允许以患者为中心的芯片，肌腱成纤维细胞将用于创建肌腱水凝胶并重新编程供体匹配的IPSC以得出内皮细胞和巨噬细胞。额外的创新将是标签的整合 - 将光子传感器带入微流体设备以允许片上传感器，长期以来一直认为对芯片机构设备的关键，未满足的需求。 UG3研究将使用芯片来验证疾病模型中的MTOR并确定与生物学相关的生物标志物。在UH3阶段，我们将使用芯片作为测试FDA批准MTOR抑制剂的效率和安全性的临床前试验平台修改药物的疾病，作为识别和优先级更安全，更有效的药物筛查平台 MTOR的抑制剂和肌腱损伤的临床试验抑制剂。成功完成这一创新项目，我们组建了一个由肌腱组织工程和手术专家组成的团队，免疫学，IPSC技术，GMP细胞制造，纳米和微型制造，传感器技术以及高吞吐量筛选。拟议的研究将开发人类的微生物生理系统来催化急性和慢性肌腱损伤的临床试验和加速药物发现。