Investigation of the molecular mechanisms of vascular endothelial dysfunction in Hutchinson-Gilford Progeria Syndrome through in vitro 2D and 3D models

通过体外2D和3D模型研究Hutchinson-Gilford早衰综合症血管内皮功能障碍的分子机制

• 批准号: 10321677
• 负责人: Yantenew G Gete
• 金额: $ 7.16万
• 依托单位: UNIV OF MARYLAND, COLLEGE PARK
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321677
• 关键词: 3-Dimensional; Adenine; Affect; Age; Aging; Arterial Fatty Streak; Atherosclerosis; Biological Assay; Biological Availability; Blood Vessels; Blood flow; Cardiovascular Diseases; Cardiovascular Pathology; Cardiovascular system; Cause of Death; Cell Culture Techniques; Cell physiology; Cells; Cessation of life; Child; Coronary Arteriosclerosis; Defect; Disease; Dominant-Negative Mutation; Endothelial Cells; Endothelium; Exhibits; Genes; Genomic DNA; Homeostasis; Human; In Vitro; Individual; Investigation; Knowledge; Lamin Type A; Lead; Lentivirus; Liquid substance; Mediating; Molecular; Mutation; NOS3 gene; Nitric Oxide; Nuclear; Olives - dietary; Organ; Pathogenesis; Patients; Pharmaceutical Preparations; Phenotype; Physiologic pulse; Physiological; Point Mutation; Population; Premature aging syndrome; Production; Progeria; Property; Proteins; Role; Smooth Muscle Myocytes; Source; Stroke; Syndrome; System; Testing; Therapeutic; Tissue Model; Toxic effect; Transgenic Mice; Tube; Umbilical vein; Variant; Vascular Endothelial Cell; base editor; cardiovascular health; endothelial dysfunction; endothelial stem cell; in vitro Model; induced pluripotent stem cell; insight; mouse model; mutant; normal aging; novel strategies; novel therapeutics; overexpression; premature; rare genetic disorder; shear stress; three-dimensional modeling; vascular endothelial dysfunction

Abstract Hutchinson-Gilford progeria syndrome (HGPS) is a rare, genetic disorder with features of accelerated aging. The majority of HGPS cases caused by a de novo point mutation in the LMNA gene (c.1824C > T; p.G608G) that results in progerin, a toxic lamin A protein variant. Children with the disease mostly die from coronary artery diseases or strokes at an average age of 14.6 years. Endothelial dysfunction is a key contributor to the cardiovascular pathobiology as the endothelium maintains vascular homeostasis and vascular tone by activating eNOS responsible for nitric oxide (NO) production. Perturbation of eNOS activity causes many diseases including atherosclerosis. Despite the vast knowledge of endothelial dysfunction in the pathogenesis of cardiovascular disease, very little known about the role of progerin in the disruption of endothelial cell function in HGPS. Furthermore, there is accumulated evidence about the role of progerin in many aspects of generalized aging and cardiovascular health. Particularly, the atherosclerotic plaques in HGPS are similar to those found in aging individuals. Moreover, vascular stiffening in HGPS is much like that seen on normal aging that manifested in both populations by increased pulse wave velocity. Using human induced pluripotent stem cell-derived endothelial cells (iPSC-ECs), I demonstrated the reduction of endothelial nitric oxide synthase (eNOS) expression level and activity in HGPS ECs compared to their normal controls. Consequently, the depletion of nitric oxide bioavailability in HGPS ECs both in static and fluidic culture conditions. Remarkably, iPSC-derived HGPS ECs exhibited eNOS dependent functional defects in forming microvascular networks that validated through over-expression of progerin in healthy human umbilical vein endothelial cells (HUVECs). I also found that Adenine Base Editor (ABEmax) that mediates the conversion of A×T to G×C in genomic DNA efficiently corrected the HGPS mutation, and the progerin expression was significantly reduced to the basal level. In addition, ABEmax rescued the nuclear blebbing phenotype of the HGPS iPSC-ECs. Thus, the study provides valuable insights into HGPS cardiovascular pathology and cardiovascular diseases associated with normal aging, and may lead to novel strategies to treat cardiovascular disease in HGPS.

抽象的 哈钦森-吉尔福德早衰综合症 (HGPS) 是一种罕见的遗传性疾病，其特征为 大多数 HGPS 病例是由 LMNA 中的从头突变引起的。 产生早老蛋白（一种有毒的儿童核纤层蛋白 A 蛋白变体）的基因（c.1824C > T；p.G608G）。 患有这种疾病的人大多死于冠状动脉疾病或中风，平均年龄为14.6岁 内皮功能障碍是心血管病理学的一个关键因素。 内皮通过激活负责的 eNOS 维持血管稳态和血管张力 eNOS 活性的干扰会导致许多疾病，包括 尽管人们对动脉粥样硬化的发病机制中的内皮功能障碍有大量了解。 心血管疾病，人们对早老素在破坏内皮细胞中的作用知之甚少 此外，关于早老素在 HGPS 中的作用的证据也越来越多。 广义衰老和心血管健康的许多方面，特别是动脉粥样硬化。 HGPS 中的斑块与老年人中发现的斑块相似。此外，血管硬化。 HGPS 与正常衰老过程中看到的情况非常相似，在两个人群中都表现为增加 脉搏波速度。 使用人类诱导多能干细胞衍生的内皮细胞 (iPSC-EC)，我证明了 HGPS 中内皮一氧化氮合酶 (eNOS) 表达水平和活性的降低 ECs 与正常对照相比，一氧化氮生物利用度的消耗。 值得注意的是，iPSC 衍生的 HGPS EC 在静态和流体培养条件下均存在。 在形成微血管网络时表现出依赖于 eNOS 的功能缺陷，验证了这一点 通过在健康人脐静脉内皮细胞（HUVEC）中过度表达早老蛋白。 我还发现腺嘌呤碱基编辑器 (ABEmax) 可以介导 A×T 到 G×C 的转换 基因组DNA有效纠正了HGPS突变，早老素表达量 显着降低至基础水平此外，ABEmax 挽救了核起泡。 因此，该研究为 HGPS 提供了有价值的见解。 心血管病理学和与正常衰老相关的心血管疾病，并且可能 导致 HGPS 治疗心血管疾病的新策略。