Epigenetic control of smooth muscle cell phenotype during microvascular remodeling

微血管重塑过程中平滑肌细胞表型的表观遗传控制

• 批准号: 10589813
• 负责人: Delphine Gomez
• 金额: $ 36.42万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10589813
• 关键词: Address; Affect; Age Years; Aging; Amputation; Area; Arteries; Back; Binding; Blood; Blood Vessels; Blood capillaries; Blood flow; Cell Communication; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cell physiology; Cells; Chimeric Proteins; DNA; DNA Methylation; Data; Defect; Development; Diameter; Disease; Docking; Embryonic Development; Endothelial Cells; Engineering; Enzymes; Epigenetic Process; Event; Excision; Gangrene; Gene Activation; Gene Expression; Genes; Genetic Transcription; Goals; Histones; Hypoxia; In Vitro; Individual; Investigation; Investments; Ischemia; KDM1A gene; Ligation; Lower Extremity; Lysine; MYH11 gene; Maintenance; Mediating; Memory; Modeling; Muscle; Pathogenesis; Patients; Perfusion; Peripheral arterial disease; Phenotype; Play; Process; Reperfusion Therapy; Repression; Risk Factors; Role; Site; Smooth Muscle Myocytes; System; Testing; Therapeutic; Tissues; Ulcer; Vascular Endothelial Growth Factors; Vascular remodeling; Vascularization; angiogenesis; arteriole; ascending aorta; cell dedifferentiation; claudication; demethylation; density; epigenome editing; improved; in vivo; migration; myocardin; novel; novel therapeutic intervention; novel therapeutics; prevent; promoter; recruit; therapeutic angiogenesis; transcription factor; Address; Affect; Age Years; Aging; Amputation; Area; Arteries; Back; Binding; Blood; Blood Vessels; Blood capillaries; Blood flow; Cell Communication; Cell Differentiation process; Cell Lineage; Cell Proliferation; Cell physiology; Cells; Chimeric Proteins; DNA; DNA Methylation; Data; Defect; Development; Diameter; Disease; Docking; Embryonic Development; Endothelial Cells; Engineering; Enzymes; Epigenetic Process; Event; Excision; Gangrene; Gene Activation; Gene Expression; Genes; Genetic Transcription; Goals; Histones; Hypoxia; In Vitro; Individual; Investigation; Investments; Ischemia; KDM1A gene; Ligation; Lower Extremity; Lysine; MYH11 gene; Maintenance; Mediating; Memory; Modeling; Muscle; Pathogenesis; Patients; Perfusion; Peripheral arterial disease; Phenotype; Play; Process; Reperfusion Therapy; Repression; Risk Factors; Role; Site; Smooth Muscle Myocytes; System; Testing; Therapeutic; Tissues; Ulcer; Vascular Endothelial Growth Factors; Vascular remodeling; Vascularization; angiogenesis; arteriole; ascending aorta; cell dedifferentiation; claudication; demethylation; density; epigenome editing; improved; in vivo; migration; myocardin; novel; novel therapeutic intervention; novel therapeutics; prevent; promoter; recruit; therapeutic angiogenesis; transcription factor

PROJECT SUMMARY Peripheral Artery Disease (PAD) is an occlusive disease of the lower extremity arteries leading to debilitating complications (e.g. claudication, amputation) due to defect in proper vascularization and efficient vascular remodeling. Recent attempts to promote therapeutic angiogenesis by VEGF therapies in patients with PAD have failed, perhaps because these strategies have only not targeted smooth muscle cells (SMC), cells that are essential for remodeling of capillaries and terminal arterioles (i.e. arteriogenesis) to increase blood distribution to ischemic regions. Arteriogenesis relies on the ability of SMC to be plastic and undergo a reversible phenotypic switching where they transiently downregulate their contractile apparatus, participate in capillary investment, and then re-differentiate back to the contractile state. However, the understanding of the mechanisms controlling SMC ability to re-differentiate and the retention of their lineage memory in vivo is limited. We previously identified an epigenetic signature specific to the SMC lineage consisting of the dimethylation of the lysine 4 of histone 3 (H3K4me2) on the promoters of the SMC marker genes (referred as SMC-H3K4me2) that is retained during SMC dedifferentiation. These observations suggest that SMC-H3K4me2 could play a pivotal role in maintenance of SMC identity and retention of lineage memory and could be a key mechanism controlling SMC participation in arteriogenesis. Studies in this proposal will test the central hypothesis that H3K4me2 controls the SMC differentiation state and enables SMC involvement in collateral capillary investment and muscularization during arteriogenesis by promoting the recruitment of TET2 on SMC marker genes. Consistent with this hypothesis, our preliminary studies, utilizing a novel locus-specific epigenome editing system to remove specifically H3K4me2 on the SMC marker genes, provide evidence that SMC-H3K4me2 tightly regulates SMC differentiation state and show that H3K4me2 interacts with Ten-eleven translocation (TET2), a key enzyme mediating DNA demethylation and a master regulator of SMC differentiation. Our central hypothesis will be further tested by addressing the following specific aims. Aim 1 will determine the impact of loss of SMC-H3K4me2 on SMC participation to arteriogenesis. The functional consequences of SMC-H3K4me2 removal on SMC phenotype, their ability to invest capillaries, as well as tissue reperfusion will be evaluated. Aim 2 will test the hypothesis that H3K4me2 plays a key role in promoting SMC differentiation by serving as a docking site for TET2 on the SMC marker genes. We expect the completion of these studies will lead to the identification of novel mechanisms controlling smooth muscle cell identity, plasticity, and lineage memory and their participation to beneficial microvascular arterialization and maturation. These results could serve from the development of new strategies for enhancing therapeutic vascularization in patients with PAD.

项目摘要 外围动脉疾病（PAD）是下肢动脉的闭塞性疾病 由于适当的血管化和有效的血管缺陷，并发症（例如，laudation，截肢） 重塑。最近在PAD患者中促进VEGF疗法促进治疗性血管生成的尝试 失败了，也许是因为这些策略仅针对平滑肌细胞（SMC），所以 重塑毛细血管和末端动脉（即小动脉生成）至关重要的要增加血液分布 到缺血区域。动脉生成依赖于SMC是塑性和经历可逆表型的能力 切换他们瞬时下调其收缩设备，参与毛细管投资和 然后重新分配回收缩州。但是，对控制机制的理解 SMC重新分化和在体内保持谱系记忆的保留的能力是有限的。我们以前已经确定 由组蛋白3的赖氨酸4的二甲基化组成的SMC谱系特有的表观遗传特征 （H3K4ME2）在SMC标记基因的启动子上（称为SMC-H3K4ME2），该基因在 SMC去分化。这些观察结果表明，SMC-H3K4ME2可以在维护中起关键作用 SMC身份和谱系记忆的保留，可能是控制SMC参与的关键机制 在动脉生成中。该提案中的研究将测试H3K4ME2控制SMC的中心假设 分化状态并使SMC参与附带毛细管投资和肌肉化期间 通过在SMC标记基因上促进TET2的募集来进行动脉生成。与这个假设一致 初步研究，利用新型基因座特异性表观基因组编辑系统去除特定的H3K4ME2 在SMC标记基因上，提供了SMC-H3K4ME2紧密调节SMC分化状态和 证明H3K4ME2与十个时期易位（TET2）相互作用，这是一种介导DNA脱甲基化的键酶 以及SMC差异化的主调节器。我们的中心假设将通过解决 遵循特定目标。 AIM 1将确定SMC-H3K4ME2损失对SMC参与的影响 动脉生成。 SMC-H3K4ME2在SMC表型上删除的功能后果，它们的能力 将评估投资毛细血管以及组织再灌注。 AIM 2将检验H3K4ME2的假设 通过在SMC标记上充当TET2的对接站点，在促进SMC差异化方面起关键作用 基因。我们预计这些研究的完成将导致确定控制的新机制 平滑肌细胞的身份，可塑性和谱系记忆及其参与有益的微血管 动脉化和成熟。这些结果可以从制定增强的新策略中起作用 PAD患者的治疗性血管化。