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.

项目概要 适当大小的血管腔的形成或管状发生是血管形成过程中的关键步骤之一 血管发育。许多血管疾病，包括静脉畸形，都与畸形有关。 或扩大的管腔。然而，我们对调节血管的分子事件仍然了解有限。 流明大小。 Abelson (Abl) 激酶信号传导调节发育和疾病期间的多种过程，包括 细胞形态发生、细胞运动、粘附和极性所需的细胞骨架重组。 Abl 信号传导 不同的细胞类型可以诱导 Rho GTPases 的激活，这是已知的管腔形成的关键调节因子。 然而，Abl 信号在调节血管腔大小中的作用此前尚未被研究过。 含有E(She)蛋白的Src同源2结构域最初被鉴定为高度保守的 因子，与 Abl 激酶相互作用。我们之前已经证明 She 具体表达在 斑马鱼胚胎中的胚胎脉管系统。然而，其在任何生物体中的生物学功能仍然未知。 在这里，我们获得了初步数据，表明她充当血管腔的新型调节剂 尺寸。功能缺陷的斑马鱼突变胚胎显示背侧血管腔扩大 主动脉。同样，缺乏 SHE 功能的人类血管内皮细胞会形成扩大的管。我们的 初步数据进一步表明 She 作为 Abl 信号传导的新型调节器，并认为 ABL 人类静脉畸形（VM）中的信号传导和管腔形成被错误调节，这表明潜在的 ABL 和 SHE 在人类疾病中的作用。我们假设 SHE 是一种新型接头蛋白，其功能 在 ABL 激酶信号通路中，在血管生成过程中限制管腔大小。我们进一步 假设 SHE 过度表达可能会减少 VM 的管腔尺寸，从而产生一种新的治疗方法。 提出以下具体目标： 1）确定细胞和分子机制 她调节肾小管发生； 2) 确定 She 是否通过抑制 Abl 激酶信号传导来限制管腔大小 途径； 3) 确定 She 过度表达是否可以减少 VM 中的管腔大小。斑马鱼胚胎缺乏 将分析其功能以发现血管生成中的细胞和分子缺陷。保护她 功能将在人类血管内皮细胞中进行测试。 Abl 激酶信号在肾小管发生中的作用 及其与 She 的相互作用将使用化学抑制剂、基因突变体和生化测定进行分析 在斑马鱼胚胎和人类细胞中。 SHE 缩小血管腔尺寸的治疗潜力 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。 获得的结果将确定 SHE 和 ABL 信号在正常和病理性肾小管发生过程中的作用， 这可能会导致治疗血管畸形的新策略的发展。