Spatial patterning modulates tissue revascularization and regeneration

空间模式调节组织血运重建和再生

• 批准号: 10053944
• 负责人: Karina Nakayama
• 金额: $ 24.9万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-20 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10053944
• 关键词: 3-Dimensional; Amputation; Anastomosis - action; Angioplasty; Architecture; Arteriographies; Autologous; Award; Balloon Angioplasty; Biological; Biological Assay; Blood Vessels; Blood capillaries; Blood flow; Cardiovascular system; Cell Culture Techniques; Cells; Cellular Morphology; Computer Interface; Coupled; Cues; Data Science; Deterioration; Diagnosis; Differentiation Antigens; Disease; Endothelial Cells; Endothelium; Engineering; Ensure; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix; FGF2 gene; First Independent Research Support and Transition Awards; Focal Adhesion Kinase 1; Foundations; Future; Gene Silencing; Generations; Genes; Genetic Transcription; Goals; Grant; Growth Factor; Histologic; Image; In Vitro; Individual; Injury; Insulin-Like Growth Factor I; Integrins; Intervention; Ischemia; Isolated limb perfusion; Isolectin; Lasers; Lead; Limb structure; Link; MAPK7 gene; Mediating; Medicine; Methodology; Mitogen-Activated Protein Kinases; Molecular; Monitor; Morbidity - disease rate; Motor Activity; Mus; Muscle; Muscle Fibers; Muscle function; Myoblasts; Natural regeneration; Operative Surgical Procedures; PECAM1 gene; Pathway interactions; Pattern; Peripheral arterial disease; Phenotype; Physiological; Platelet-Derived Growth Factor; Play; Process; Property; Public Health; Reconstructive Surgical Procedures; Recovery of Function; Reperfusion Therapy; Research; Role; Running; Sampling; Series; Signal Transduction; Site; Skeletal Muscle; Skeletal Muscle Myosins; Stains; Stents; Structure; Therapeutic; Therapeutic Intervention; Tissue Engineering; Tissues; Training; Transplantation; Treatment Efficacy; Tube; United States; United States National Institutes of Health; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; Vascularization; Vein graft; Work; angiogenesis; artery occlusion; base; blood perfusion; cadherin 5; career; critical limb Ischemia; cytokine; density; disability; efficacy evaluation; externship; functional genomics; health goals; implantation; improved; injured; innovation; insight; loss of function; mechanical properties; medical schools; mortality; mouse model; muscle physiology; muscle regeneration; myogenesis; nanofibrillar; nanoscale; neutralizing antibody; novel; paracrine; regenerative; repaired; restoration; scaffold; therapeutic evaluation; tissue injury; tissue regeneration; transcriptome sequencing; vascular injury; von Willebrand Factor

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.

项目概要 美国有 850 万人患有外周动脉疾病 (PAD)。由于疾病 进展时，可能会导致四肢动脉血流严重受阻，从而导致危重肢体 缺血，并且与从最初开始仅 6 个月内高达 20% 的极高死亡率相关 诊断。这种情况需要立即进行血管内治疗以重新建立血流，通常 通过使用支架、球囊血管成形术或自体静脉移植物；然而，这些治疗需要 多种干预措施并不能最终降低截肢率。治疗干预措施旨在 长期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。该 K99/R00 独立之路奖建立在之前的工作基础之上，展示了 来自纳米级细胞外基质的空间图案线索调节内皮细胞（EC）形态 和血管生成功能。当前研究的目标是使用对齐的 3D 纳米级细胞引导 增强血管内皮细胞的血管生成潜力的支架，其再生目标是恢复血液 流向缺血区域并实现严重受损组织的功能修复，这是一项重要的公共卫生 一直难以实现的目标。 首先，该奖项将提供机会来检验使用对齐的空间图案的作用 与非图案支架相比，在增强 EC 血管生成功能以及调节 肌肉成肌细胞表型和机械特性。与此目标同时，治疗效果 与非图案化支架相比，EC 接种对齐支架将进行组织评估 体积肌肉和血管损伤小鼠模型中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 EC 表型并同时解决 血管生成反过来会增强肌肉肌纤维的分化和成熟。最后，为了更深入地了解 了解基因网络和途径协同作用以促进的机制 通过空间模式的血管生成、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途径。拟议的培训将包括通过以下方式提供的课程： 斯坦福大学医学院和心血管医学领域顶尖专家的实习机会， 数据科学和肌肉再生。拟议的系列研究将加深对 空间细胞模式增强 EC 血管生成的生物学机制 肌肉成肌细胞功能。这些研究的结果将为未来的再生提供见解 严重受损和缺血组织血运重建的策略和工程治疗方法，以及 将成为治疗广泛血管疾病的创新平台和重要一步。