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 形成适当尺寸或微管发生的血管腔是在 血管发展。包括静脉畸形在内的许多血管疾病与畸形有关 或放大的流明。但是，我们对调节血管的分子事件的理解仍然有限 管腔尺寸。 Abelson（ABL）激​​酶信号传导调节发育和疾病期间的各种过程，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信号在中 不同的细胞类型可以诱导Rho GTPases的激活，Rho GTPase是腔形成的关键调节剂。 但是，ABL信号在调节血管管腔大小的调节中的作用尚未进行。 SRC同源性2含有E（SHE）蛋白的结构域最初被识别为高度保守的 因子，与ABL激酶相互作用。我们以前已经证明她在 斑马鱼胚胎中的胚胎脉管系统。但是，其生物学功能在任何生物体中仍然尚不清楚。 在这里，我们获得了初步数据，该数据认为她是血管腔的新型调节剂 尺寸。斑马鱼突变体胚胎缺乏SHE功能表现出的背侧血管腔的扩大 主动脉。同样，人血管内皮细胞缺乏SHE功能，形成了扩大的管。我们的 进一步的数据进一步表明她是ABL信号的新型调节剂，并认为ABL 信号传导和管腔形成在人静脉畸形（VM）中被误导，表明潜力 ABL和她在人类疾病中的作用。我们假设她是一种新型的衔接蛋白，功能 在ABL激酶信号传导途径中，在血管肾小管发生过程中限制管腔大小。我们进一步 假设她的过表达可能会减小VM中的管腔大小，从而导致一种新型的治疗方法。 提出了以下特定目的：1）确定细胞和分子机制 她调节了肾变生成； 2）确定她是否通过抑制ABL激酶信号来限制管腔尺寸 途径； 3）确定她的过表达是否可以减小VM中的管腔尺寸。斑马鱼胚胎缺乏 她的功能将分析血管肾小管发生中的细胞和分子缺损。她的保护 功能将在人血管内皮细胞中进行测试。 ABL激酶信号在微管发生中的作用 并将使用化学抑制剂，基因突变体和生化测定法对其进行分析 在斑马鱼胚胎和人类细胞中。她减少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。 获得的结果将确定在正常和病理管状发生过程中SHE和ABL信号传导的作用， 这可能导致发展血管畸形的新型策略。