Spatial patterning modulates tissue revascularization and regeneration

项目摘要 8.5在美国有100万人患有外周动脉疾病（PAD）。随着疾病 进展，它可以导致动脉血流到四肢的严重阻塞， 缺血，并与高达20%的死亡率非常高，从最初的6个月 诊断.这种情况需要立即进行血管内治疗以重建血流，通常 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 多次干预，并不能最终降低截肢率。治疗干预措施， 长期的功能恢复必须增强组织血管生成，同时恢复生理功能， 组织结构这个K99/R 00独立之路奖建立在以前的工作，证明 来自纳米细胞外基质的空间模式提示调节内皮细胞（EC）形态 和血管生成功能。本研究的目的是使用来自对齐3D的纳米级细胞引导， 支架，以增强血管内皮细胞的血管生成潜力，其再生目标是恢复血液 血液流向缺血区域，使严重受损的组织能够进行功能性修复，这是一项重要的公共卫生 这是一个很难实现的目标。 首先，该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血管生成功能以及调节 肌成肌细胞表型和机械性能。与此同时，研究了 与非图案化支架相比，将评估EC接种的对齐支架的组织 在体积性肌肉和血管损伤的小鼠模型中进行血管再生和肌肉再生。 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化在多个方面解决问题，以诱导伴随着 血管生成，这将反过来增强肌肉肌纤维分化和成熟。最后，为了获得更深层次的 了解基因网络和途径协同工作的机制，以促进 血管生成通过空间模式，在基因沉默和功能基因组学的方法将是 用于揭示新的细胞模式化途径。拟议的培训将包括通过以下途径提供的课程： 斯坦福大学医学院和心血管医学领域的顶尖专家， 数据科学和肌肉再生拟议的一系列研究将加深对 空间细胞模式化赋予EC血管生成增强的生物学机制， 肌肉成肌细胞功能。这些研究的结果将提供见解，为未来的再生 用于严重受损和缺血组织的血运重建的策略和工程治疗，以及 将成为治疗多种血管疾病的创新平台和重要步骤。

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.