Research Project 2

项目摘要 跟腱病是一个非常普遍和昂贵的临床问题。然而，目前的手术和药物 肌腱修复的策略是有限的，治疗疾病的非手术策略集中在刺激肌腱 通过物理治疗进行修复。因此，改善肌腱治疗策略的临床需求尚未得到满足 受伤肌腱退化改变了化学物理环境，改变了生物物理输入， 细胞（tenocytes）。肌腱细胞的正常和异常表型都是由动态空间分布来定义的。 他们的基因组的时间组织，因此，重要的是要了解三维基因组架构， 跟腱细胞的变化与跟腱病和化学机械线索如何调节转录和 在变性细胞中的染色质谱，以开发更好的治疗策略，肌腱病。此外，委员会认为， 在退化环境中负责肌腱表型变化的表观遗传机制是 探索不足表观遗传药物是可用的，并且已用于治疗目的，并且可能还 构成了通过操纵表观遗传景观治疗肌腱病的有前途的途径 和肌腱细胞的3D染色质结构以锁定适当的细胞表型。为了解决这些悬而未决的问题， 研究项目的总体目标是检验我们的假设，即跟腱病改变了表观遗传学， 景观，3D染色质结构，和肌腱细胞中影响其表型的转录特征， 这些改变可以通过生物物理学线索和表观遗传学的结合来操纵和恢复。 修饰语拟议的工作是重要的，因为它将产生新的知识，如何改变机械 疾病谱中的负荷和机械信号传导影响基因组组织和肌腱细胞 表型，以及这些变化如何定义疾病进展和治疗干预。我们的目标是： 目的1：确定跟腱病如何改变纳米染色质组织和可及性 肌腱细胞的景观，影响它们的表型。目的2：确定生物物理线索和表观遗传 修饰剂在“退化”肌腱细胞中恢复“健康”肌腱细胞基因组组织以改善治疗效果 战略布局拟议的研究是创新的，因为我们将使用尖端的全基因组和单细胞 分析研究，第一次，跟腱病如何调节纳米染色质状态， 转录活性，使用基于单细胞的成像和测序技术。这些研究将确定 跟腱病理和疾病发作的新表观遗传机制，新的机械负荷 范式和小表观基因组修饰分子，提供关键和新颖的信息，以支持新的 机械表观遗传策略，以提高有针对性的物理治疗方案的疗效。

PROJECT SUMMARY Achilles tendinopathy is a very prevalent and costly clinical problem. However, current surgical and drug strategies for tendon repair are limited, and non-surgical strategies to treat disease focus on stimulating tendon repair through physical therapy. Thus, there is an unmet clinical need to improve treatment strategies for tendon injuries. Tendon degeneration alters the chemo-physical environment and changes biophysical inputs to resident cells (called tenocytes). Both normal and aberrant phenotypes in tendon cells are defined by the dynamic spatio- temporal organization of their genome, and so it will be important to understand how 3D genome architecture in tendon cells changes with Achilles tendinopathy and how chemo-mechanical cues regulate transcriptional and chromatin profiles in degenerative cells to develop better therapeutic strategies for tendinopathies. Furthermore, the epigenetic mechanisms responsible for the tendon phenotype change in degenerative environments are underexplored. Epigenetic drugs are available and have been used for therapeutic purposes and likely also constitute a promising avenue for treatment of tendinopathies through manipulation of the epigenetic landscape and 3D chromatin architecture of tendon cells to lock in proper cell phenotype. To address these open questions, the overall goal of Research Project is to test our hypotheses that Achilles tendinopathy alters epigenetic landscape, 3D chromatin architecture, and transcriptional signatures in tenocytes impacting their phenotype, and that these alterations can be manipulated and restored via the combination of biophysical cues and epigenetic modifiers. The proposed work is significant as it will generate new knowledge about how changes in mechanical loading and mechano-signaling across the spectrum of disease impacts genome organization and tendon cell phenotype, and how these changes define disease progression and therapeutic interventions. Our Aims are: Aim 1: Determine how Achilles tendinopathy alters the nanoscale chromatin organization and accessibility landscape of tenocytes, impacting their phenotype. Aim 2: Identify whether biophysical cues and epigenetic modifiers restore ‘healthy’ tenocyte genome organization in ‘degenerative’ tenocytes to improve therapeutic strategies. The proposed research is innovative as we will use cutting-edge genome wide and single cell analyses to study, for the first time, how Achilles tendinopathy regulates nanoscale chromatin states and transcriptional activity, using single-cell based imaging and sequencing technologies. These studies will identify novel epigenetic mechanisms of Achilles tendon pathology and disease onset, new mechanical loading paradigms, and small epigenome-modifying molecules, providing critical and novel information to support new mechano-epigenetic strategies to improve the efficacy of targeted physical therapy protocols.