Advanced Tissue Engineered Models of Human Cartilage for Studying Joint Disease

项目摘要/摘要 关节软骨的疾病和创伤极大地削弱了生活质量，因为受损的软骨具有 自我修复的能力非常有限。关节软骨缺损的处理仍然是一种普遍的和 治疗选择有限，具有挑战性的问题，部分原因是缺乏高保真模式 促进我们对疾病/损伤进展的理解，并使新疗法的测试成为可能。啮齿动物是 通常用于研究关节疾病和发育，但它们未能概括人类的关键方面 软骨解剖学和生物学，特别是人类软骨复杂的带状组织。在设计过程中 体外模型在生物学上是可靠的，它们通常缺乏体内模型提供的复杂性，包括 与其他组织的相互作用。为了解决这一差距，我提议设计出带状组织的人类软骨 在体外，通过施加适当的时空梯度的转化生长因子-β信号(目标1)，同时 包括用于增强骨软骨相互作用的活的软骨下骨(目标2)。我试图证明 利用单基因结缔组织疾病研究关节疾病的人软骨模型的应用， 马凡综合征，其中纤维蛋白缺陷影响转化生长因子-β的生物利用度和肌肉骨骼发育 (目标3)。我假设转化生长因子-β信号的精确调控和软骨下骨的包含 基质将生成具有带状组织和软骨-骨的类自然的人类关节软骨 相互作用，这可以用来模拟疾病，如MFS。光遗传学提出了一种策略，通过它我们可以 可以通过光以前所未有的精度控制转化生长因子-β信号。在骨软骨交界处，生化 生物力学的相互作用调节软骨的健康和疾病，然而大多数已建立的软骨模型都失败了 由于支持这两种组织类型的复杂性，因此需要结合骨骼。我们的实验室已经设计了灌流 生物反应器可以为每种组织类型提供单独的物理和化学线索，以克服这一点 挑战。使用MFS患者来源的HiPSCs，先进的工程化软骨模型将被用于 概述疾病的功能和结构特征，以临床数据为基准，并与数据进行比较 来自现有的软骨源性微团培养的体外模型。拟议的工作将建立一部小说 为软骨组织工程提供了先进的人体组织模型，为关节疾病的研究提供了先进的组织模型。

PROJECT SUMMARY / ABSTRACT Disease and trauma of articular cartilage are highly debilitating to the quality of life, as damaged cartilage has a very limited ability for self-repair. The management of articular cartilage defects continues to be a prevalent and challenging problem with limited treatment options, in part due to the lack of high-fidelity models that would advance our understanding of disease/injury progression and enable testing of novel therapeutics. Rodents are commonly used to study joint disease and development, yet they fail to recapitulate key aspects of human cartilage anatomy and biology, in particular the intricate zonal organization of human cartilage. While engineered in vitro models can be biologically faithful, they generally lack the complexity offered by in vivo models, including the interactions with other tissues. To address this gap, I propose to engineer zonally organized human cartilage in vitro by applying the appropriate spatiotemporal gradients of TGF-β signaling (Aim 1), while concurrently including living subchondral bone for enhanced osteochondral interactions (Aim 2). I seek to demonstrate the utility of this human cartilage model for studying joint disease using a monogenic connective tissue disorder, Marfan syndrome (MFS), in which fibrillin defects affect TGF-β bioavailability and musculoskeletal development (Aim 3). I hypothesize that the precise regulation of TGF-β signaling and the inclusion of a subchondral bone substrate will generate native-like human articular cartilage with zonal organization and cartilage-bone interactions, which can be used to model diseases such as MFS. Optogenetics presents a strategy by which we can control TGF-β signaling by light with unprecedented precision. At the osteochondral junction, biochemical and biomechanical interactions mediate cartilage health and disease, yet most established cartilage models fail to incorporate bone due to the complexity of supporting both tissue types. Our lab has designed perfusion bioreactors which can provide separate physical and chemical cues to each tissue type to overcome this challenge. Using MFS patient-derived hiPSCs, the advanced engineered cartilage model will be used to recapitulate functional and structural features of disease, benchmarked to clinical data and compared to data from existing in vitro models of chondrogenic micromass cultures. The proposed work will establish a novel method for cartilage tissue engineering and provide an advanced human tissue model for studying joint diseases.