Endothelial cell autonomous mechanisms of blood vessel diameter control

血管直径控制的内皮细胞自主机制

• 批准号: 10421065
• 负责人: Arndt Friedrich Siekmann
• 金额: $ 43.56万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-06-15 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10421065
• 关键词: 1-Phosphatidylinositol 3-Kinase; Actomyosin; Address; Affect; Apical; Basal Cell; Biological Assay; Biomechanics; Blood; Blood Vessels; Body part; Caliber; Cell Count; Cell Polarity; Cell Shape; Cell Size; Cells; Conceptions; Cytoskeleton; Data; Defect; Development; Dimensions; Disease; Dominant-Negative Mutation; Embryo; Embryonic Development; Endoglin; Endothelial Cells; Endothelium; Event; Failure; Feedback; Genes; Human; Immunohistochemistry; Knowledge; Lead; Measures; Membrane; Molecular; Mutation; Myosin Light Chain Kinase; Nutrient; Oxygen; PARD6A gene; PIK3CA gene; Pathway interactions; Patients; Pattern; Pharmacology; Phenotype; Property; Proteins; Protocols documentation; Reporter; Reporting; Shapes; Shunt Device; Signal Transduction; Smooth Muscle Myocytes; Testing; Time; Transforming Growth Factor beta; Transgenic Organisms; Tumor Suppressor Proteins; Zebrafish; apical membrane; basolateral membrane; cell dimension; constriction; curative treatments; early onset; imaging approach; insight; malformation; membrane polarity; mutant; overexpression; prevent; targeted treatment

Abstract Blood vessels are important for the distribution of nutrients and oxygen to all parts of our bodies. A hierarchical organization of differently sized blood vessels is key for their functionality. Several diseases can affect blood vessel topologies and diameters, leading to vascular malformations in humans. Previous reports suggested that increases in blood vessel diameters can be caused by increases in endothelial cell numbers, the cells of the blood vessels’ inner lining. We established an imaging approach in zebrafish embryos that allows us to precisely analyze endothelial cell numbers in addition to their shapes and sizes during embryonic development. Surprisingly, we found that initially the shapes and sizes of endothelial cells were more critical for blood vessel diameter control than their numbers and that endothelial cell numbers increased only at later stages of vascular malformation establishment. Importantly, we also found that endothelial shapes and sizes were affected in mutants of several different genes causing vascular malformations. However, to date it is not known how these genes affect cell shapes and sizes and how this would impact blood vessel diameters. The aims in this proposal address this question by analyzing the cellular and molecular components influencing cell shapes and sizes. In aim 1 we will investigate how the cytoskeleton and its contractile properties affect cellular dimensions and how this might feedback on blood vessel diameters. We will also investigate whether changing the cytoskeleton and hence endothelial cell contractility can rescue vascular malformations. In aim 2 we plan to interrogate the influence of endothelial cell polarity on blood vessel diameters. Endothelial cells have an apical, facing the blood vessel lumen, and a basolateral membrane domain. At present, we do not know how mutations causing vascular malformations change apical-basal polarity and how these changes would affect blood vessel diameters. We will test for these possibilities by examining apical and basolateral polarity in different zebrafish mutants that develop vascular malformations. We will also investigate how changing apical-basolateral polarity will affect endothelial cell shapes and sizes in these mutants. Ultimately, we aim to reverse vascular malformations through normalizing apical-basolateral polarity and endothelial cell contractility and thereby endothelial cell shapes prior to increases in endothelial cell numbers.

抽象的 血管对于将营养和氧气分配到我们身体的各个部位很重要。分层的 不同大小血管的组织对其功能至关重要。多种疾病都会影响血液 血管拓扑和直径，导致人类血管畸形。此前的报道表明 血管直径的增加可能是由于内皮细胞数量的增加引起的。 血管的内壁。我们在斑马鱼胚胎中建立了一种成像方法，使我们能够精确地 分析胚胎发育过程中内皮细胞的数量以及形状和大小。 令人惊讶的是，我们发现最初内皮细胞的形状和大小对于血管的形成更为关键 直径控制大于其数量，并且内皮细胞数量仅在血管形成的后期阶段增加 畸形建立。重要的是，我们还发现内皮形状和大小受到影响 导致血管畸形的几种不同基因的突变体。然而，迄今为止，尚不清楚这些 基因影响细胞的形状和大小，以及这将如何影响血管直径。本提案的目标 通过分析影响细胞形状和大小的细胞和分子成分来解决这个问题。在 目标 1 我们将研究细胞骨架及其收缩特性如何影响细胞尺寸以及如何 这可能会反馈血管直径。我们还将研究是否改变细胞骨架和 因此，内皮细胞的收缩力可以挽救血管畸形。在目标 2 中，我们计划询问 内皮细胞极性对血管直径的影响。内皮细胞有一个顶端，面向血液 血管腔和基底外侧膜域。目前，我们还不知道突变是如何引起血管的 畸形会改变顶端-基底极性以及这些变化如何影响血管直径。我们将 通过检查不同斑马鱼突变体的顶端和基底外侧极性来测试这些可能性 血管畸形。我们还将研究改变顶端-基底外侧极性将如何影响内皮细胞 这些突变体的细胞形状和大小。最终，我们的目标是通过正常化来逆转血管畸形 顶端-基底外侧极性和内皮细胞收缩性，从而增加之前的内皮细胞形状 在内皮细胞数量中。