Spatial patterning modulates tissue revascularization and regeneration

项目总结 美国有850万人患有外周动脉疾病(PAD)。作为一种疾病 进展时，可导致流向四肢的动脉血流严重阻塞，造成危重肢体。 脑缺血，与最初6个月高达20%的死亡率有关 诊断。这种情况通常需要立即进行血管内治疗以重建血液流动。 通过使用支架、球囊血管成形术或自体静脉移植；然而，这些治疗需要 多项干预措施并不能最终降低截肢率。针对以下目标的治疗干预 长期功能恢复必须在恢复生理功能的同时促进组织血管生成 组织架构。这个K99/R00独立之路奖建立在之前的工作基础上，证明了 来自纳米级细胞外基质的空间构图信号调节内皮细胞(EC)的形态 和血管生成功能。目前这项研究的目标是使用来自排列的3D的纳米级细胞引导 增强血管内皮细胞血管生成潜能的支架，其再生目标是恢复血液 流向缺血区并使严重受损组织能够进行功能性修复，这是一项重要的公共健康 这是一个具有挑战性的目标。 首先，这一奖项将提供机会来研究使用对齐的空间图案的作用 与非图案化支架相比，在增强EC血管生成功能和调节血管生成方面的作用 肌成肌细胞表型和力学特性。在实现这一目标的同时， EC种子排列支架与非图案化支架相比，将对组织进行评估 容积性肌肉和血管损伤小鼠模型的血运重建和肌肉再生。 通过这些研究，恢复受损组织的血管和肌肉功能的挑战是 通过使用空间细胞图案来诱导伴随着 血管生成，进而促进肌肉肌纤维的分化和成熟。最后，为了获得更深层次的 对基因网络和途径协同工作以促进 通过空间模式、基因沉默和功能基因组学方法实现的血管生成 用来揭示新的细胞图案化途径。拟议的培训将包括通过 斯坦福医学院和心血管医学领域的顶尖专家的外部奖学金， 数据科学和肌肉再生。拟议的一系列研究将加深对 空间细胞构图增强EC血管生成和血管生成的生物学机制 肌肉成肌细胞的功能。这些研究的发现将提供洞察力，为未来的再生 用于严重受损和缺血组织血运重建的策略和工程疗法；以及 将成为治疗各种血管疾病的创新平台和重要一步。

PROJECT SUMMARY 8.5 million people in the United States suffer from peripheral arterial disease (PAD). As the disease progresses, it can lead to severe obstruction of arterial blood flow to the extremities causing critical limb ischemia, and is associated with devastatingly high mortality rates of up to 20% just 6 months from initial diagnosis. This condition requires immediate endovascular treatment to re-establish blood flow, commonly through the use of stents, balloon angioplasty, or autologous vein grafts; however, these treatments require multiple interventions and do not conclusively lower the amputation rates. Therapeutic interventions aimed at long-term functional recovery must augmenting tissue angiogenesis concomitant with restoring physiological tissue architecture. This K99/R00 Pathway to Independence Award builds on previous work that demonstrates that spatial patterning cues from nanoscale extracellular matrices modulate endothelial cell (EC) morphology and angiogenic function. The objective of the current study is to use nanoscale cell guidance from aligned 3D scaffolds to enhance the angiogenic potential of vascular ECs, with the regenerative goal of restoring blood flow to ischemic regions and enabling functional repair of severely damaged tissue, an important public health goal that has been challenging to attain. First, this award will provide the opportunity to examine the role of spatial patterning using aligned versus non-patterned scaffolds, in the enhancement of EC angiogenic function as well as the modulation of muscle myoblasts phenotype and mechanical properties. In parallel with this aim, the therapeutic efficacy of EC-seeded aligned scaffolds in comparison to non-patterned scaffolds, will be assessed for tissue revascularization and muscle regeneration in a mouse model of volumetric muscle and vascular injury. Through these studies, the challenge of restoring both vascular and muscular function to injured tissues is tackled on multiple fronts by using spatial cell patterning to induce an EC phenotype concomitant with angiogenesis that will in turn enhance muscle myofiber differentiation and maturation. Finally, to gain a deeper understanding of the mechanisms by which gene networks and pathways work in concert to promote angiogenesis through spatial patterning, methodologies in gene silencing and functional genomics will be employed to reveal novel cell patterning pathways. The proposed training will include courses offered through the Stanford School of Medicine and externships with leading experts in the fields of cardiovascular medicine, data science, and muscle regeneration. The proposed series of studies will deepen the understanding of the biological mechanisms through which spatial cell patterning confers enhancement of EC angiogenesis and muscle myoblast function. Findings from these studies will provide insights that will inform future regenerative strategies and engineered therapeutics for revascularization of severely damaged and ischemic tissues, and will serve as an innovative platform and important step in the treatment of a broad range of vascular diseases.