Role of SHE and ABL signaling in vascular tubulogenesis

SHE 和 ABL 信号在血管生成中的作用

• 批准号: 10587279
• 负责人: Saulius Sumanas
• 金额: $ 63.94万
• 依托单位: UNIVERSITY OF SOUTH FLORIDA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-03-01 至 2026-11-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10587279
• 关键词: Adaptor Signaling Protein; Adhesions; Aorta; Biochemical; Biological Assay; Biological Process; Biomedical Engineering; Blood Vessels; Cell Culture Techniques; Cell Separation; Cells; Chemicals; Clinical; Cytoskeletal Modeling; Cytoskeleton; Data; Defect; Development; Diameter; Disease; Dorsal; E protein; Embryo; Endothelial Cells; Event; Family; Genetic; Growth; Human; Knowledge acquisition; Lesion; Maintenance; Modeling; Molecular; Morphogenesis; Mus; Natural regeneration; Organism; Pathologic; Patients; Phosphotransferases; Play; Process; Protein Tyrosine Kinase; Proteins; Role; Signal Pathway; Signal Transduction; Testing; Therapeutic; Tube; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelium; Venous Malformation; Xenograft Model; Xenograft procedure; Zebrafish; cell motility; cell type; experimental study; fluid flow; human disease; improved; inhibitor; malformation; mutant; novel; novel strategies; novel therapeutic intervention; overexpression; regenerative therapy; repaired; rho GTP-Binding Proteins; three dimensional cell culture; tool; vascular bed

Project Summary Formation of vascular lumen of appropriate size, or tubulogenesis, is one of critical steps during vascular development. Many vascular diseases including venous malformations are associated with malformed or enlarged lumens. However, we still have a limited understanding of molecular events that regulate vascular lumen size. Abelson (Abl) kinase signaling regulates diverse processes during development and disease, including cytoskeletal reorganization required for cell morphogenesis, cell motility, adhesion and polarity. Abl signaling in different cell types can induce activation of Rho GTPases, which are known key regulators of lumen formation. However, the role of Abl signaling in regulating vascular lumen size has not been previously investigated. Src homology 2 domain containing E (She) protein was originally identified as a highly conserved factor, which interacted with Abl kinase. We have previously demonstrated that She is specifically expressed in embryonic vasculature in zebrafish embryos. However, its biological function is still unknown in any organism. Here we obtained preliminary data which argues that She acts as a novel regulator of vascular lumen size. Zebrafish mutant embryos deficient in she function display enlarged vascular lumen within the dorsal aorta. Similarly, human vascular endothelial cells, deficient in SHE function, form enlarged tubes. Our preliminary data further suggest that She functions as a novel regulator of Abl signaling, and argue that ABL signaling and lumen formation are misregulated in human venous malformation (VM), suggesting a potential role for ABL and SHE in human disease. We hypothesize that SHE is a novel adaptor protein which functions in the ABL kinase signaling pathway to restrict lumen size during vascular tubulogenesis. We further hypothesize that SHE overexpression may reduce lumen size in VM leading to a novel therapeutic approach. The following specific aims are proposed: 1) Determine the cellular and molecular mechanisms by which She regulates tubulogenesis; 2) Determine if She restricts lumen size by inhibiting Abl kinase signaling pathway; 3) Determine if She overexpression can reduce lumen size in VM. Zebrafish embryos deficient in she function will be analyzed for cellular and molecular defects in vascular tubulogenesis. Conservation of She function will be tested in human vascular endothelial cells. The role of Abl kinase signaling in tubulogenesis and its interaction with She will be analyzed using chemical inhibitors, genetic mutants and biochemical assays in zebrafish embryos and human cells. The therapeutic potential of SHE to reduce the size of vascular lumen will be analyzed in the cell culture and mouse VM model as well as in primary cells isolated from VM patients. Obtained results will identify the role for SHE and ABL signaling during normal and pathological tubulogenesis, which may lead to the development of novel strategies to treat vascular malformations.

项目摘要 适当大小的血管腔的形成，或管发生，是在血管形成过程中的关键步骤之一。 血管发育包括静脉畸形在内的许多血管疾病都与畸形 或扩大的管腔。然而，我们对调节血管生成的分子事件的了解仍然有限。 管腔尺寸。 Abelson（Abl）激酶信号传导调节发育和疾病期间的多种过程，包括 细胞骨架重组是细胞形态发生、细胞运动、粘附和极性所需的。Abl信号转导 不同的细胞类型可以诱导RhoGTP酶的活化，RhoGTP酶是已知的管腔形成的关键调节剂。 然而，Abl信号传导在调节血管腔大小中的作用先前尚未被研究。 含Src同源2结构域的E（She）蛋白最初被鉴定为高度保守的蛋白质， 因子，其与Abl激酶相互作用。我们以前已经证明，她是专门表达在 斑马鱼胚胎中的胚胎脉管系统。然而，它的生物学功能在任何生物体中仍然是未知的。 在这里，我们获得了初步的数据，认为她作为一个新的调节血管腔 尺寸she功能缺陷的斑马鱼突变胚胎显示背部血管腔扩大 主动脉类似地，缺乏SHE功能的人血管内皮细胞形成扩大的管。我们 初步数据进一步表明She作为Abl信号传导的一种新的调节剂发挥作用，并认为ABL 在人类静脉畸形（VM）中，信号传导和管腔形成失调，这表明潜在的 ABL和SHE在人类疾病中的作用。我们推测SHE是一种新的衔接蛋白， 在ABL激酶信号通路中，以限制血管小管形成过程中的管腔大小。我们进一步 假设SHE过表达可以减小VM中管腔尺寸，从而导致新的治疗方法。 提出了以下具体目标：1）确定细胞和分子机制， 2）确定She是否通过抑制Abl激酶信号传导来限制管腔大小 3）确定She过表达是否可以减小VM中的管腔尺寸。斑马鱼胚胎缺乏 她的功能将被分析为在血管小管发生中的细胞和分子缺陷。保护她 将在人血管内皮细胞中测试功能。Abl激酶信号在肾小管形成中的作用 将使用化学抑制剂、遗传突变体和生化测定来分析它与She的相互作用 在斑马鱼胚胎和人类细胞中。SHE缩小血管腔的治疗潜力 将在细胞培养物和小鼠VM模型以及从VM患者分离的原代细胞中进行分析。 所获得的结果将确定SHE和ABL信号传导在正常和病理性小管发生中的作用， 这可能导致治疗血管畸形的新策略的发展。