Gene Regulatory Mechanisms that Repress BMP2 in Pathological Calcification

病理钙化中抑制 BMP2 的基因调控机制

• 批准号: 9216823
• 负责人: MELISSA B ROGERS
• 金额: $ 39.73万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-15 至 2020-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9216823
• 关键词: 3' Untranslated Regions; Adult; Age; Aging; Alleles; Amputation; Angioplasty; Aorta; Arteries; Atherosclerosis; Calcified; Calcinosis; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cells; Cessation of life; Chronic Kidney Failure; Collection; Complement; Conserved Sequence; Coronary Arteriosclerosis; Dangerousness; Data; Diabetes Mellitus; Disease; Elderly; Elements; Event; Functional disorder; Gene Targeting; Genetic Transcription; Genetically Engineered Mouse; Goals; Heart Diseases; Heart Valves; Human; Impairment; Kidney; Kidney Failure; Knockout Mice; LacZ Genes; Lead; Mediating; Mesenchymal; Messenger RNA; Metabolism; MicroRNAs; Minerals; Modeling; Molecular; Mus; Muscle Cells; Mutant Strains Mice; Normal tissue morphology; Outcome; Pathologic; Pathology; Physiology; Premature aging syndrome; Protein Biosynthesis; Publishing; Reagent; Regulator Genes; Renal function; Reporter; Repression; Research; Research Design; Signal Transduction; Stroke; Testing; Therapeutic; Time; Tissues; Transcription Repressor/Corepressor; Transgenes; United States; Update; Vascular Smooth Muscle; Vascular calcification; age related; aged; aortic valve; aortic valve disorder; bone cell; bone morphogenetic protein 2; calcification; calcium phosphate; gene repression; microRNA biomarkers; novel; novel therapeutics; osteogenic; pre-clinical; prevent; research clinical testing; restenosis; soft tissue; statistics; therapy design; 3' Untranslated Regions; Adult; Age; Aging; Alleles; Amputation; Angioplasty; Aorta; Arteries; Atherosclerosis; Calcified; Calcinosis; Cardiovascular Diseases; Cardiovascular system; Cause of Death; Cells; Cessation of life; Chronic Kidney Failure; Collection; Complement; Conserved Sequence; Coronary Arteriosclerosis; Dangerousness; Data; Diabetes Mellitus; Disease; Elderly; Elements; Event; Functional disorder; Gene Targeting; Genetic Transcription; Genetically Engineered Mouse; Goals; Heart Diseases; Heart Valves; Human; Impairment; Kidney; Kidney Failure; Knockout Mice; LacZ Genes; Lead; Mediating; Mesenchymal; Messenger RNA; Metabolism; MicroRNAs; Minerals; Modeling; Molecular; Mus; Muscle Cells; Mutant Strains Mice; Normal tissue morphology; Outcome; Pathologic; Pathology; Physiology; Premature aging syndrome; Protein Biosynthesis; Publishing; Reagent; Regulator Genes; Renal function; Reporter; Repression; Research; Research Design; Signal Transduction; Stroke; Testing; Therapeutic; Time; Tissues; Transcription Repressor/Corepressor; Transgenes; United States; Update; Vascular Smooth Muscle; Vascular calcification; age related; aged; aortic valve; aortic valve disorder; bone cell; bone morphogenetic protein 2; calcification; calcium phosphate; gene repression; microRNA biomarkers; novel; novel therapeutics; osteogenic; pre-clinical; prevent; research clinical testing; restenosis; soft tissue; statistics; therapy design

The goal of this project is to understand how aging and chronic kidney disease (CKD) promote bone morphogenetic protein 2 (BMP2) synthesis that furthers pathological calcification of the heart valves and vasculature. Post-transcriptional regulatory mechanisms repress BMP2 in aorta and aortic valve. We hypothesize that (1) this repression is essential for controlling BMP2 levels in the adult and that (2) conditions such as aging and CKD impair the function of factors that mediate this repression in healthy heart valves and aorta. We will test these hypotheses in aged normal mice (24 months) and in the Klotho null mouse which models age-related disorders, including CKD. Klotho null mice suffer premature aging and death occurs at 7 - 8 weeks of age. At this time, extensive calcification of the heart valves, vasculature, and other soft tissues has occurred. AIM 1 is to test the influence of conditionally deleting a strong repressive element in the 3'untranslated region (UTR) of Bmp2 on calcification in normal aged mice and in Klotho null mice with premature aging and aging associated renal dysfunction. We will use recently developed Bmp2 alleles to assess how the deletion of this potent post-transcriptional repressor influences the course of calcification associated with aging and renal dysfunction. Preliminary results indicate that the UCS inhibits calcification. AIM 2 is to identify and compare miRNA signatures unique to young, healthy aorta and aortic valve to the signatures of these tissues from normal aged mice and in Klotho null mice with premature aging and severe vascular calcification. Within these profiles, we will focus on post- transcriptional repressive factors (miRNAs) that target the Bmp2 UCS and contribute to 3'UTR mediated repression in healthy tissues. AIM 3 is to test how selected and validated miRNAs influence the expression Bmp2 and downstream osteogenic events that lead to calcification in Klotho null mice bearing our unique transgenes. Our novel Bmp2 reporter mouse will expedite pre-clinical testing of miRNA therapies that prevent pathological calcification. Our newly developed Bmp2 allele (Aim 1) will differentiate changes due these miRNAs targeting Bmp2 relative to off-target genes. The outcomes of the proposed research will be (1) increased understanding of how BMP2 influences pathological calcification, (2) the identification and analyses of potential miRNA biomarkers, and (3) new therapeutic leads for controlling pathological calcification.

该项目的目的是了解衰老和慢性肾脏疾病（CKD）如何促进 骨形态发生蛋白2（BMP2）合成，促进了病理钙化 心脏瓣膜和脉管系统。 转录后调节机制抑制主动脉和主动脉瓣中的BMP2。我们 假设（1）这种抑制对于控制成年人的BMP2水平至关重要 （2）衰老和CKD等条件损害了调解这一点的因素的功能 在健康的心脏瓣膜和主动脉中抑制。 我们将在老年正常小鼠（24个月）和klotho null小鼠中检验这些假设 哪些模型与年龄有关的疾病，包括CKD。 klotho null小鼠过早衰老 死亡发生在7-8周龄。目前，心脏瓣膜广泛钙化， 脉管系统和其他软组织发生了。 目的1是测试有条件删除强烈压制元素的影响 BMP2在正常老年小鼠和klotho null中的钙化中的3'ununtranslated区域（UTR） 过早衰老和衰老相关的肾功能障碍的小鼠。我们最近将使用 开发的BMP2等位基因，以评估该有效的转录后阻遏物的删除 影响与衰老和肾功能障碍相关的钙化过程。初步的 结果表明，UCS抑制钙化。 AIM 2是识别和比较年轻，健康主动脉和主动脉独特的miRNA签名 瓣膜到正常老年小鼠和klotho null小鼠的阀 过早衰老和严重的血管钙化。在这些配置文件中，我们将专注于职位 针对BMP2 UCS并有助于3'UTR的转录抑制因子（miRNA） 健康组织中介导的抑制作用。 AIM 3是测试选择和验证的miRNA如何影响BMP2的表达和 下游的成骨事件导致klotho null小鼠的钙化，带有我们独特的 转基因。我们的新型BMP2报告基因鼠标将加快miRNA的临床前测试 预防病理钙化的疗法。我们新开发的BMP2等位基因（AIM 1）将 由于这些靶向BMP2相对于靶向基因的miRNA而区分变化。 拟议研究的结果将是（1）对BMP2的了解增加 影响病理钙化，（2）潜在miRNA的识别和分析 生物标志物和（3）新的治疗铅用于控制病理钙化。