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

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.

项目概要 外周动脉疾病（PAD）是一种下肢动脉闭塞性疾病，会导致患者衰弱 由于适当的血管化和有效的血管缺陷导致的并发症（例如跛行、截肢） 重塑。最近尝试通过 VEGF 疗法促进 PAD 患者的治疗性血管生成 失败了，也许是因为这些策略没有针对平滑肌细胞（SMC），而平滑肌细胞是 对于毛细血管和终末小动脉的重塑（即动脉生成）以增加血液分布至关重要 到缺血区域。动脉生成依赖于 SMC 的可塑性和可逆表型的能力 切换他们短暂下调收缩装置的位置，参与毛细血管投资，以及 然后重新分化回收缩状态。然而，对控制机制的理解 SMC 在体内重新分化和保留谱系记忆的能力是有限的。我们之前确定的 SMC 谱系特有的表观遗传特征，由组蛋白 3 的赖氨酸 4 的二甲基化组成 （H3K4me2）位于 SMC 标记基因（称为 SMC-H3K4me2）的启动子上，该基因在 SMC去分化。这些观察结果表明 SMC-H3K4me2 可以在维护中发挥关键作用 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。该提案中的研究将检验 H3K4me2 控制 SMC 的中心假设 分化状态并使 SMC 参与侧支毛细血管投资和肌肉化 通过促进 SMC 标记基因上 TET2 的募集来促进动脉生成。与这个假设相一致，我们的 初步研究，利用新型位点特异性表观基因组编辑系统特异性去除 H3K4me2 SMC标记基因，提供SMC-H3K4me2严格调节SMC分化状态的证据 表明 H3K4me2 与 10-11 易位 (TET2) 相互作用，TET2 是介导 DNA 去甲基化的关键酶 以及 SMC 分化的主要调节因子。我们的中心假设将通过解决以下问题得到进一步检验： 遵循特定目标。目标 1 将确定 SMC-H3K4me2 丢失对 SMC 参与的影响 动脉生成。 SMC-H3K4me2 去除对 SMC 表型的功能影响及其能力 将评估毛细血管投资以及组织再灌注。目标 2 将检验 H3K4me2 的假设 作为 SMC 标记上 TET2 的对接位点，在促进 SMC 分化中发挥关键作用 基因。我们预计这些研究的完成将导致识别新的控制机制 平滑肌细胞特性、可塑性和谱系记忆及其对有益微血管的参与 动脉化和成熟。这些结果可以用于制定新的战略来增强 PAD 患者的治疗性血管化。

项目成果

Nobody Is Perfect: Cre Drivers Deserve Careful Consideration.

没有人是完美的：Cre 驱动程序值得仔细考虑。

• DOI: 10.1161/atvbaha.123.319683
• 发表时间: 2023
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Gomez,Delphine

Translational opportunities of single-cell biology in atherosclerosis.

• DOI: 10.1093/eurheartj/ehac686
• 发表时间: 2023-04-07
• 期刊: EUROPEAN HEART JOURNAL
• 影响因子: 39.3
• 作者: de Winther, Menno P. J.;Back, Magnus;Evans, Paul;Gomez, Delphine;Goncalves, Isabel;Jorgensen, Helle F.;Koenen, Rory R.;Lutgens, Esther;Norata, Giuseppe Danilo;Osto, Elena;Dib, Lea;Simons, Michael;Stellos, Konstantinos;Yla-Herttuala, Seppo;Winkels, Holger;Bochaton-Piallat, Marie-Luce;Monaco, Claudia
• 通讯作者: Monaco, Claudia