A microphysiological system of tendon inflammation and fibrosis for drug screening and efficacy testing: MPS Database Engagement

用于药物筛选和功效测试的肌腱炎症和纤维化的微生理系统：MPS 数据库参与

• 批准号: 10430792
• 负责人: Hani A Awad
• 金额: $ 7.54万
• 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 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10430792
• 关键词: Address; Animal Model; Award; Biological Models; Blood Vessels; Blood coagulation; Cell Culture Techniques; Cells; Characteristics; Cicatrix; Clinical; Clinical Trials; Collagen; Communities; Companions; Connective Tissue; Consensus; Database Management Systems; Databases; Development; Devices; Disease; Drug Screening; Enzyme-Linked Immunosorbent Assay; Evaluation; FDA approved; FRAP1 gene; Feedback; Fibrosis; Funding; Gel; Gene Expression; Generations; Genetic; Human; Immune; In Situ; Infiltration; Inflammation; Inflammatory; Intervention; Measurable; Measurement; Modeling; Morphology; Mus; Myofibroblast; Operative Surgical Procedures; Parents; Pathology; Patients; Phenotype; Play; Procedures; Process; Protocols documentation; Reproducibility; Research Design; Role; Safety; System; Tendon Injuries; Tendon structure; Time; Tissues; Universities; Vascular Endothelial Cell; base; cytokine; design; drug efficacy; efficacy evaluation; efficacy testing; follow-up; healing; human disease; human model; induced pluripotent stem cell; injury and repair; mTOR Inhibitor; microphysiology system; monocyte; neovascularization; novel; photonics; pre-clinical; pre-clinical research; repaired; response; senescence; sensor; simulation; success; temporal measurement; tissue injury; tool; virtual clinical trial

Abstract The continued advancement of microphysiological systems (MPS) as pre-clinical research tools is vital to overcome the low throughput and inaccuracies inherent in animal models of human disease. The limitations of pre-clinical animal models, most commonly mice, are particularly apparent in inflammatory diseases which are known to have distinct genetic and cytokine responses to inflammation. The establishment of MPS alternatives, however, will require scientific consensus on the protocols and systems best suited to address particular diseases. As the current MPS era is characterized by a proliferation of approaches, the Microphysiological System Data Base (MPS-db) created by the University of Pittsburgh is a valuable tool to hasten the development of MPS standards. Because the success of the MPS-db requires the active participation by MPS developers and users, we seek supplemental funding to contribute the designs, protocols and results for an MPS system that models the interplay between inflammation and fibrosis in tendon healing (UG3TR00287). Importantly, the injury and repair of connective tissue injury is not represented in the current MPS-db but accounts for more than 8.5 million clinical procedures annually, including 2 million major surgeries. Our human tendon-on-a-chip (hToC) model focuses on the early inflammatory stages of tendon repair, where timely interventions may promote scarless healing. The hToC features vascular and collagen compartments which exchange soluble and cellular factors in a simulation of the neovascularized microenvironment established shortly after blood clotting. Monocyte infiltration is hypothesized to play an essential role in the generation of contractile myofibroblasts which progress to senescence and release monocyte activating factors in a positive feedback loop that causes scar tissue. The model uses iPSCs derived from primary human tenocytes to create vascular endothelial cells and monocytes in an isogenic, patient-centric triculture. With supplemental funding we will share: 1) descriptions of the mechanisms of the tendon injury and fibroinflammatory repair process; 2) design details for the hToC including device components and modules for both flow and integrated photonic- based sensing; 3) cell culture and device protocols including phenotypic characteristics and operational parameters such as flow rates for priming of ECs and the introduction of immune cells; 4) The design and rationale for studies under baseline and inflammation/repair conditions; and 5) Results including an analysis of intra-study reproducibility.

抽象的 微生理系统（MPS）作为临床前研究工具的持续发展对于 克服人类疾病动物模型固有的低通量和不准确性。的局限性 临床前动物模型，最常见的是小鼠，在炎症性疾病中尤其明显，这些疾病 已知对炎症具有独特的遗传和细胞因子反应。建立 MPS 替代方案， 然而，需要就最适合解决特定问题的协议和系统达成科学共识 疾病。由于当前 MPS 时代的特点是方法的激增，微生理学 匹兹堡大学创建的系统数据库（MPS-db）是加速 制定 MPS 标准。因为MPS-db的成功需要MPS的积极参与 开发人员和用户，我们寻求补充资金来贡献设计、协议和结果 MPS 系统模拟肌腱愈合过程中炎症和纤维化之间的相互作用 (UG3TR00287)。 重要的是，结缔组织损伤的损伤和修复并未在当前的 MPS-db 中体现，但 每年临床手术量超过850万例，其中大型手术200万例。我们的人类 肌腱芯片（hToC）模型专注于肌腱修复的早期炎症阶段，及时 干预措施可能会促进无疤痕愈合。 hToC 具有血管和胶原隔室， 在建立的新生血管微环境模拟中交换可溶性和细胞因子 血液凝固后不久。假设单核细胞浸润在单核细胞的产生中发挥重要作用 收缩性肌成纤维细胞逐渐衰老并释放单核细胞激活因子 导致疤痕组织的反馈回路。该模型使用源自人类原代肌腱细胞的 iPSC 来创建 以患者为中心的同基因三培养中的血管内皮细胞和单核细胞。通过补充资金，我们 将分享：1）肌腱损伤机制和纤维炎症修复过程的描述； 2） hToC 的设计细节，包括用于流动和集成光子的设备组件和模块 基于传感； 3) 细胞培养和设备方案，包括表型特征和操作 参数，例如 EC 启动的流速和免疫细胞的引入； 4）设计和 在基线和炎症/修复条件下进行研究的基本原理； 5) 结果包括分析 研究内的再现性。

项目成果

Preclinical tendon and ligament models: Beyond the 3Rs (replacement, reduction, and refinement) to 5W1H (why, who, what, where, when, how).

临床前肌腱和韧带模型：超越 3R（替换、减少和细化）到 5W1H（为什么、谁、什么、何地、何时、如何）。

• DOI: 10.1002/jor.25678
• 发表时间: 2023
• 期刊: Journal of orthopaedic research : official publication of the Orthopaedic Research Society
• 作者: Little,Dianne;Amadio,PeterC;Awad,HaniA;Cone,StephanieG;Dyment,NathanielA;Fisher,MatthewB;Huang,AliceH;Koch,DrewW;Kuntz,AndrewF;Madi,Rashad;McGilvray,Kirk;Schnabel,LaurenV;Shetye,SnehalS;Thomopoulos,Stavros;Zhao,Chunf
• 通讯作者: Zhao,Chunf

The Modular µSiM: A Mass Produced, Rapidly Assembled, and Reconfigurable Platform for the Study of Barrier Tissue Models In Vitro.

• DOI: 10.1002/adhm.202200804
• 发表时间: 2022-09
• 期刊: ADVANCED HEALTHCARE MATERIALS
• 影响因子: 10
• 作者: McCloskey, Molly C.;Kasap, Pelin;Ahmad, S. Danial;Su, Shiuan-Haur;Chen, Kaihua;Mansouri, Mehran;Ramesh, Natalie;Nishihara, Hideaki;Belyaev, Yury;Abhyankar, Vinay V.;Begolo, Stefano;Singer, Benjamin H.;Webb, Kevin F.;Kurabayashi, Katsuo;Flax, Jonathan;Waugh, Richard E.;Engelhardt, Britta;McGrath, James L.
• 通讯作者: McGrath, James L.

A computer vision approach for analyzing label free leukocyte trafficking dynamics on a microvascular mimetic.

• DOI: 10.3389/fimmu.2023.1140395
• 发表时间: 2023
• 期刊: Frontiers in immunology
• 影响因子: 7.3
• 作者: Ahmad SD;Cetin M;Waugh RE;McGrath JL
• 通讯作者: McGrath JL

NF-κB activation persists into the remodeling phase of tendon healing and promotes myofibroblast survival.

• DOI: 10.1126/scisignal.abb7209
• 发表时间: 2020-11-17
• 期刊: Science signaling
• 影响因子: 7.3
• 作者: Best KT;Nichols AEC;Knapp E;Hammert WC;Ketonis C;Jonason JH;Awad HA;Loiselle AE
• 通讯作者: Loiselle AE

In vitro Studies of Transendothelial Migration for Biological and Drug Discovery.

• DOI: 10.3389/fmedt.2020.600616
• 发表时间: 2020
• 期刊: Frontiers in medical technology
• 作者: Salminen AT;Allahyari Z;Gholizadeh S;McCloskey MC;Ajalik R;Cottle RN;Gaborski TR;McGrath JL
• 通讯作者: McGrath JL