Research Project 2

项目总结 跟腱病是一个非常普遍和昂贵的临床问题。然而，目前的外科和药物 肌腱修复的策略是有限的，而非手术治疗疾病的策略侧重于刺激肌腱。 通过理疗进行修复。因此，改善肌腱的治疗策略是临床上尚未得到满足的需要。 受伤。肌腱退变改变了化学物理环境，改变了生物物理输入 细胞(称为腱细胞)。肌腱细胞的正常表型和异常表型都是由动态的空间分布决定的。 它们基因组的时间组织，因此了解3D基因组结构如何在 跟腱病变时腱细胞的变化以及化学机械信号如何调节转录和 变性细胞中的染色质图谱，以开发更好的肌腱病治疗策略。此外， 退行性环境中肌腱表型改变的表观遗传机制是 开发不足。表观遗传药物是可用的，并已用于治疗目的，很可能还 构成了通过操纵表观遗传景观来治疗肌腱病的一条有希望的途径 肌腱细胞的3D染色质结构，以锁定适当的细胞表型。为了解决这些未解决的问题， 研究项目的总体目标是测试我们的假设，即跟腱病变改变表观遗传学 肌腱细胞的景观、3D染色质结构和转录特征影响其表型，以及 这些变化可以通过生物物理线索和表观遗传学的组合来操纵和修复 修饰符。这项拟议的工作意义重大，因为它将产生关于机械结构如何变化的新知识 跨疾病谱的负载和机械信号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： 目标1：确定跟腱病变如何改变纳米级染色质的组织和可获得性 肌腱细胞的景观，影响其表型。目标2：确定生物物理线索和表观遗传学 修饰剂可以恢复退行性肌腱细胞中的健康肌腱细胞基因组组织，以提高治疗效果 战略。拟议的研究是创新的，因为我们将使用先进的全基因组和单细胞 首次分析研究跟腱病如何调节纳米级染色质状态和 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 跟腱病理和疾病发生的新表观遗传学机制，新的机械负荷 范例和小表观基因组修饰分子，提供关键和新的信息来支持新的 机械-表观遗传策略，以提高靶向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.