Pharmacologic PTEN upregulation: A novel therapeutic approach to prevent pathological vascular remodeling and fibrosis

药理学 PTEN 上调：预防病理性血管重塑和纤维化的新治疗方法

• 批准号: 9756685
• 负责人: Keith Strand
• 金额: $ 3.21万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-10 至 2021-05-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9756685
• 关键词: Acetylation; Affect; Angiotensin II; Animal Model; Arterial Fatty Streak; Biological Assay; Blood Vessels; Bromodomain; Cardiovascular Diseases; Cardiovascular Models; Cause of Death; Cell Differentiation process; Cell Nucleus; Cells; Chromatin; Complex; Connective Tissue; Coronary artery; Country; Cytometry; Data; Development; Disease Progression; Dose; EP300 gene; Enzyme-Linked Immunosorbent Assay; Epigenetic Process; Family; Fibrosis; Fluorescence; Gene Expression; Generations; Genetic; Genetic Transcription; Growth Factor; Heterogeneity; Human; Hyperplasia; Immune; Impairment; In Vitro; Inflammation; Inflammatory; Injury; Knockout Mice; Knowledge; Label; Mediating; Microscopy; Modeling; Mus; PTEN gene; Pathogenicity; Pathologic; Perivascular Fibrosis; Pharmacology; Phenotype; Phosphoric Monoester Hydrolases; Play; Process; Proteins; Reader; Regulation; Reporter; Research; Role; Serum Response Factor; Severities; Signal Transduction; Smooth Muscle Myocytes; Stimulus; System; T-Lymphocyte; Testing; Therapeutic; Therapeutic Effect; Tissue Sample; Transcriptional Regulation; Tumor Suppressor Genes; Tumor Suppressor Proteins; United States; Up-Regulation; Vascular Smooth Muscle; Vascular remodeling; Work; base; cardiovascular disorder therapy; cell type; chemokine; clinically relevant; cytokine; gene repression; histological stains; in vitro activity; in vivo; inflammatory marker; inhibitor/antagonist; knock-down; macrophage; member; mouse model; myocardin; new therapeutic target; novel; novel therapeutic intervention; overexpression; p65; pre-clinical; prevent; promoter; protective effect; recruit; response; response to injury; single-cell RNA sequencing; small molecule; therapeutic evaluation; transcription factor

Project Summary/Abstract Cardiovascular disease is the leading cause of death in the United States and is marked by the development of pathological vascular remodeling which contributes to disease progression. In response to injury or pathological stimuli, resident vascular smooth muscle cells (SMCs) contribute to these remodeling processes by de- differentiating from a quiescent, contractile phenotype towards a proliferative, pro-inflammatory, pro-fibrotic phenotype capable of recruiting immune cells like macrophages and T-cells. The preliminary data supporting this proposal indicates genetic, systemic upregulation of the potent tumor suppressor PTEN maintains SMC differentiation, reduces perivascular immune cell recruitment and blunts remodeling in multiple pre-clinical animal models of cardiovascular disease. This suggests that pharmacologic PTEN upregulation could hold significant therapeutic potential in treating cardiovascular disease by inhibiting vascular remodeling. Currently, the therapeutic potential of PTEN is not fully exploited as there are few compounds known to increase PTEN expression. To fill this knowledge gap, a recently completed high-throughput compound screen (HTS) of over 2,500 compounds using a fluorescence-based PTEN promoter-reporter system was used to identify 11 hit compounds with dose-responsive ability to induce PTEN promoter activity. Recently, JQ1, an inhibitor of the epigenetic reader protein Brd4 that is known to have anti-remodeling effects, was shown to upregulate PTEN. However, preliminary data presented in this proposal suggest that the anti-remodeling effects of JQ1 are blunted in PTEN depleted SMCs both in vitro and in vivo, suggesting that the protective effects of JQ1 are mediated through PTEN upregulation. Little is known about the role of Brd4, the target of JQ1, in regulating PTEN levels in SMCs. However, Brd4 has been shown to interact directly with the histone acetyltransferase p300 and the p65 subunit of the pro-inflammatory transcription factor NFkB, both of which are known to affect PTEN expression. This application presents a novel model of PTEN regulation where competition for limited amounts of p300 drives either PTEN expression through promoter acetylation, or inflammatory gene expression through p300-NFkB interaction mediated by Brd4. Specific Aim 1 will expand upon the preliminary data and will use in vitro and in vivo PTEN depletion to determine whether the vasculoprotective effects of JQ1 are due to increased PTEN expression mediated by increased p300 activity at the PTEN promoter and inhibition of Brd4-p300-NFkB complex formation. Specific Aim 2 will extend the results of the completed HTS to test hit compounds in vitro and in vivo for their ability to induce PTEN expression and reduce vascular remodeling, focusing specifically on how PTEN upregulation affects crosstalk between SMCs and recruited macrophages/T-cells. To summarize, these studies will explore a potentially novel mechanism regulating PTEN expression which may play an essential role in mediating the known anti-remodeling effects of JQ1 and will identify novel compounds targeting PTEN that could be developed as new translational therapies for cardiovascular disease.

项目总结/摘要 在美国，心血管疾病是导致死亡的主要原因， 病理性血管重塑，其有助于疾病进展。针对损伤或病理性 刺激，常驻血管平滑肌细胞（SMC）有助于这些重塑过程，通过去- 从静止的收缩表型向增殖的促炎的促纤维化的表型分化， 表型能够招募免疫细胞，如巨噬细胞和T细胞。初步数据支持 这一建议表明，基因，系统性上调的有效的肿瘤抑制因子PTEN维持SMC 在多种临床前动物中， 心血管疾病的模型。这表明，药理学PTEN上调可能具有显著的 在通过抑制血管重塑治疗心血管疾病中的治疗潜力。目前 PTEN的治疗潜力尚未完全开发，因为已知增加PTEN的化合物很少 表情为了填补这一知识空白，最近完成了一项高通量化合物筛选（HTS）， 使用基于荧光的PTEN启动子-报告子系统的2，500种化合物用于鉴定11种命中 具有诱导PTEN启动子活性的剂量响应能力的化合物。最近，JQ 1，一种抑制剂， 已知具有抗重塑作用的表观遗传阅读器蛋白Brd 4显示上调PTEN。 然而，该提案中提出的初步数据表明，JQ 1的抗重塑作用减弱 在体外和体内的PTEN缺失的SMC中，表明JQ 1的保护作用是介导的， 通过PTEN的上调关于JQ 1的靶点Brd 4在调节PTEN水平中的作用知之甚少 在SMC中。然而，Brd 4已被证明直接与组蛋白乙酰转移酶p300相互作用， 促炎性转录因子NF κ B的p65亚基，已知两者都影响PTEN 表情本申请提出了一种新的PTEN调控模型，其中竞争有限的量 p300的表达通过启动子乙酰化驱动PTEN表达，或通过启动子乙酰化驱动炎性基因表达。 Brd 4介导的p300-NFkB相互作用。具体目标1将在初步数据的基础上进行扩展，并将在 体外和体内PTEN耗竭，以确定JQ 1的血管保护作用是否是由于增加的 通过增加PTEN启动子处的p300活性和抑制Brd 4-p300-NFkB介导的PTEN表达 复杂的形成。具体目标2将扩展已完成的HTS的结果，以测试体外命中化合物 以及在体内诱导PTEN表达和减少血管重塑的能力，特别关注 PTEN上调如何影响SMC和募集的巨噬细胞/T细胞之间的串扰。总而言之， 这些研究将探索一种潜在的调节PTEN表达的新机制， 在介导JQ 1已知的抗重塑作用中起重要作用，并将鉴定靶向 PTEN可能被开发为心血管疾病的新的转化疗法。