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 形成适当大小的管腔或小管形成是治疗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与畸形有关。 或管腔增大。然而，我们对调节血管的分子事件的了解仍然有限。 管腔大小。 Abelson(Abl)激酶信号在发育和疾病过程中调节不同的过程，包括 细胞形态发生、细胞运动、黏附和极性所需的细胞骨架重组。ABL信令传入 不同的细胞类型可以诱导Rho GTP酶的激活，Rho GTP酶是已知的管腔形成的关键调节因子。 然而，Abl信号在调节血管管腔大小中的作用还没有被研究过。 含有E(Se)蛋白的SRC同源2结构域最初被鉴定为高度保守的 与Abl激酶相互作用的因子。我们之前已经证明，她在 斑马鱼胚胎中的胚胎血管系统。然而，它的生物学功能在任何生物体中都是未知的。 在这里，我们获得了初步的数据，证明她是一种新的血管管腔调节器 尺码。SHE功能缺陷的斑马鱼突变胚胎在背部显示血管腔扩大 大动脉。类似地，缺乏SHE功能的人血管内皮细胞形成扩大的管子。我们的 初步数据进一步表明，她是ABL信号的一种新的调节者，并认为ABL 在人类静脉畸形(VM)中，信号和管腔的形成是错误的，提示存在潜在的 ABL和SHE在人类疾病中的作用。我们假设她是一种新型的接头蛋白，其功能 在ABL激酶信号通路中，在血管小管形成过程中限制管腔大小。我们进一步 假设SHE的过度表达可能会减少VM的管腔大小，从而导致一种新的治疗方法。 提出了以下具体目标：1)通过以下途径确定细胞和分子机制 2)确定她是否通过抑制abl激酶信号来限制管腔大小。 3)确定过度表达SHE是否可以缩小VM的管腔大小。斑马鱼胚胎缺乏 SHE的功能将被分析血管小管形成过程中的细胞和分子缺陷。保护SHE 功能将在人类血管内皮细胞中进行测试。Abl激酶信号转导通路在肾小管发生中的作用 并将使用化学抑制剂、基因突变和生化分析来分析其与SHE的相互作用。 在斑马鱼胚胎和人类细胞中。SHE缩小血管腔大小的治疗潜力 将在细胞培养和小鼠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信号在正常和病理性肾小管形成中的作用， 这可能会导致开发治疗血管畸形的新策略。