Novel quantitative proteomic approaches to define the altered interplay between OGlcNAcylation and Phosphorylation in myofilament dysfunction of diabetic hearts

新的定量蛋白质组学方法来定义糖尿病心脏肌丝功能障碍中 OGlcNAc 酰化和磷酸化之间相互作用的改变

• 批准号: 10004702
• 负责人: Genaro Antonio Ramirez-Correa
• 金额: $ 12.46万
• 依托单位: UNIVERSITY OF TEXAS RIO GRANDE VALLEY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10004702
• 关键词: Adrenergic Agents; Adult; Alanine; Anabolism; Aspartic Acid; Basic Science; Biomedical Research; Calcium; Cardiac; Cardiac health; Cardiology; Cardiovascular system; Child; Complication; Core Facility; Coronary Arteriosclerosis; Data; Depressed mood; Diabetes Mellitus; Diagnostic; Disease; Doctor of Philosophy; Echocardiography; Environment; Equilibrium; Etiology; Faculty; Fellowship; Foundations; Frequencies; Functional disorder; Funding; Future; Gene Transfer; Gene Transfer Techniques; Glucose; Goals; Heart; Heart Diseases; Heart failure; Hexosamines; Human; Impairment; In Vitro; Individual; Ions; K-Series Research Career Programs; Knowledge; Label; Lead; Link; Maps; Mass Spectrum Analysis; Measurement; Medical; Medical Students; Mentors; Mexican Americans; Microfilaments; Molecular; Molecular Motors; Muscle; Myocardial; Myocardial dysfunction; Myocardium; National Heart, Lung, and Blood Institute; North America; Pathway interactions; Patients; Pediatric cardiology; Phosphorylation; Phosphorylation Site; Physiological; Play; Post-Translational Protein Processing; Postdoctoral Fellow; Prevalence; Property; Proteins; Proteomics; Reporting; Research; Research Personnel; Research Proposals; Risk; Role; Sarcomeres; Scheme; Scientist; Serine; Site; Techniques; Testing; Threonine; Time; Training; Training Support; Translational Research; Translations; Troponin I; Type 2 diabetic; United States National Institutes of Health; Universities; Variant; Work; Workload; adeno-associated viral vector; career; diabetic; diabetic cardiomyopathy; diabetic patient; experience; gene transfer vector; glucose metabolism; heart function; improved; in vivo; inorganic phosphate; interest; medical schools; mitochondrial dysfunction; multiple reaction monitoring; muscle physiology; mutant; novel; novel therapeutics; pandemic disease; pediatric department; professor; response; skills; stoichiometry; success; titanium dioxide; tool

Project Abstract/Summary The candidate I am a Mexican-American MD/PhD who works as a basic scientist. I am also an Assistant Professor in the Johns Hopkins School of Medicine, Division of Pediatric Cardiology where I also completed my post- doctoral fellowship training. My interest in and commitment to a translational and basic research career started as a 4th year medical student. I am convinced that funding through the NIH/NHLBI Mentored Career Development Award to Promote Faculty Diversity in Biomedical Research will be instrumental to achieve my goal, which is to become an independent investigator and a future leader in the field of molecular cardiology and its translation into the advancement of therapies of diabetic cardiomyopathy and heart failure.  Research Proposal: Novel quantitative proteomic approaches to define the altered interplay between O-GlcNAcylation and Phosphorylation in myofilament dysfunction of diabetic hearts In North America, the 2010 prevalence of diabetes was 37.4 million (10.2%) and is on a steady rise16. Diabetic patients are 2 to 4 times more at risk of dying from heart disease than the general population17. Among cardiovascular complications, diabetic cardiomyopathy refers to a progressive diastolic and systolic dysfunction due to a contractile deficit of the cardiac muscle that develops independently from coronary artery disease. While it is present in 60% of diabetic patients, no therapy is currently available to halt or significantly alter the course of diabetic cardiomyopathy18. Post-translational modifications of the sarcomere regulate cardiac function and when dysregulated contribute to cardiac dysfunction. Recent work in our group has focused on the identification, quantification and functional characterization of myofilament O-GlcNAcylation and Phosphorylation1-8. The goal of this proposal is to use state of the art quantitative proteomic approaches to extensively map and perform site-specific quantification of all potentially O-GlcNAcylated and Phosphorylated myofilament proteins of normal and diabetic hearts during baseline cardiac function and during β-adrenergic and force-frequency stimulation. By comparing O-GlcNAc/Phosphate stoichiometry changes between baseline and enhanced workload we will identify key sites for abnormal myofilament function in diabetic cardiomyopathy. By using gene transfer techniques, the present proposal also will perform in vivo and in vitro functional work to define the role of the interplay between O-GlcNAcylation and Phosphorylation and the mechanisms that lead to impaired cardiac contractile reserve in diabetes. Advances in this field can potentially generate early diagnostic tools for diabetic cardiomyopathy and open new therapeutic venues to fix the molecular motors of a failing diabetic heart. The specific aims of this proposal are Aim 1: To perform global myofilament site-specific O- GlcNAcylation and Phosphorylation mapping and quantification in normal and type 2 diabetic hearts. Aim 2: To identify O-GlcNAcylated and Phosphorylated sites with the greatest stoichiometric variation during β-adrenergic and force-frequency stimulation in normal and type 2 diabetic heart myofilaments. Aim 3: To validate the functional impact of altered balance of O-GlcNAcylation and Phosphorylation competing sites on cardiac contractility by manipulating myofilament proteins with gene transfer. The environment The Johns Hopkins School of Medicine possesses an excellent environment to perform basic and translational research. Johns Hopkins University has a strong foundation and facilities in research focused on diseases of adults and children. For example, the Department of Pediatrics presently has 27 million dollars in NIH research dollars. The medical campus will enable the candidate to access numerous state-of-the-art core facilities. The mentors, advisors and collaborators outlined in this application will assist in a successful completion of the candidate career and research goals. We have assembled a superb team of fine scientist and established faculty with many years of experience and great success mentoring young scientists. My main mentor is Dr. Anne M Murphy, co-mentor is Dr. Jennifer Van Eyk. Dr. Gerald W. Hart and Dr. Brian O'Rourke form the advisor committee.

项目摘要/摘要 候选 我是一名墨西哥裔美国人，是一名基础科学家。我也是一名助理教授， 约翰霍普金斯医学院儿科心脏病学分部，我也在那里完成了我的博士后工作， 博士奖学金培训。我对转化和基础研究事业的兴趣和承诺始于 作为一个四年级的医学生。我相信，通过NIH/NHLBI指导职业的资金， 发展奖，以促进生物医学研究的教师多样性将有助于实现我的 我的目标是成为分子心脏病学领域的独立研究者和未来的领导者 并将其转化为糖尿病心肌病和心力衰竭治疗的进步。 研究提案：新型定量蛋白质组学方法来定义改变的相互作用 糖尿病心肌肌丝功能障碍中O-GlcNAc酰化和磷酸化的研究 在北美，2010年糖尿病患病率为3740万（10.2%），并呈稳步上升趋势16。 糖尿病患者死于心脏病的风险是普通人群的2至4倍17。 在心血管并发症中，糖尿病性心肌病是指进行性舒张和收缩功能障碍， 由于独立于冠状动脉的心肌收缩缺陷而引起的功能障碍 疾病虽然它存在于60%的糖尿病患者中，但目前没有治疗方法可以阻止或显著减少糖尿病的发生。 改变糖尿病心肌病的病程18. 肌节的翻译后修饰调节心脏功能， 会导致心脏功能障碍我们小组最近的工作集中在识别、量化和 肌丝O-GlcNAc酰化和磷酸化的功能表征1 -8.这项提案的目的是 使用最先进的定量蛋白质组学方法来广泛地绘制和执行位点特异性 对正常和正常人的所有潜在的O-GlcNAc酰化和磷酸化肌丝蛋白进行定量， 糖尿病心脏在基线心脏功能和β-肾上腺素能和力-频率刺激期间。通过 比较基线和增强工作负荷之间的O-GlcNAc/磷酸盐化学计量变化，我们将 确定糖尿病心肌病中肌丝功能异常关键部位。通过基因转移 技术，本提案还将进行体内和体外功能性工作，以确定的作用， O-GlcNAc酰化和磷酸化之间的相互作用以及导致心脏受损的机制 糖尿病的收缩储备这一领域的进展可能会产生糖尿病的早期诊断工具。 心肌病和开辟新的治疗场所，以修复 糖尿病心脏衰竭的分子马达的具体目标 这些建议是 目的1：进行全局肌丝位点特异性O- GlcNAc化和磷酸化作图和定量 正常和2型糖尿病心脏。 目的2：鉴定O-GlcNAc酰化和磷酸化位点， 在β-肾上腺素能和 力-频刺激对正常和2型糖尿病心脏的影响 肌丝 目的3：验证O-GlcNAc酰化和磷酸化竞争平衡改变的功能影响 通过基因转移操纵肌丝蛋白质对心肌收缩力的影响。 环境 约翰霍普金斯医学院拥有一个良好的环境，执行基本和 翻译研究约翰霍普金斯大学拥有强大的基础和设施，研究重点是 成人和儿童的疾病。例如，儿科部门目前有2700万美元， 美国国立卫生研究院研究经费。医学校园将使候选人能够访问许多国家的最先进的核心 设施在此应用程序中概述的导师，顾问和合作者将协助成功 完成候选人的职业生涯和研究目标。我们已经组建了一个由优秀科学家组成的优秀团队 并建立了具有多年经验的教师队伍，在指导年轻科学家方面取得了巨大成功。我的主要 导师是Anne M Murphy博士，共同导师是Jennifer货车Eyk博士。Gerald W.哈特和布莱恩·奥罗克博士 成立顾问委员会。

项目成果

Epidemiological Algorithm for Early Detection of COVID-19 Cases in a Mexican Oncologic Center.

用于早期检测墨西哥肿瘤中心的COVID-19病例的流行病学算法。

• DOI: 10.3390/healthcare10030462
• 发表时间: 2022-03-01
• 期刊: Healthcare (Basel, Switzerland)
• 作者: González-Escamilla M;Pérez-Ibave DC;Burciaga-Flores CH;Ortiz-Murillo VN;Ramírez-Correa GA;Rodríguez-Niño P;Piñeiro-Retif R;Rodríguez-Gutiérrez HF;Alcorta-Nuñez F;González-Guerrero JF;Vidal-Gutiérrez O;Garza-Rodríguez ML
• 通讯作者: Garza-Rodríguez ML

Identification of a Novel Pathogenic Rearrangement Variant of the APC Gene Associated with a Variable Spectrum of Familial Cancer.

• DOI: 10.3390/diagnostics11030411
• 发表时间: 2021-02-28
• 期刊: Diagnostics (Basel, Switzerland)
• 作者: Garza-Rodríguez ML;Treviño V;Pérez-Maya AA;Rodríguez-Gutiérrez HF;González-Escamilla M;Elizondo-Riojas MÁ;Ramírez-Correa GA;Vidal-Gutiérrez O;Burciaga-Flores CH;Pérez-Ibave DC
• 通讯作者: Pérez-Ibave DC

Risk Association of TOX3 and MMP7 Gene Polymorphisms with Sporadic Breast Cancer in Mexican Women.

• DOI: 10.3390/curroncol29020086
• 发表时间: 2022-02-11
• 期刊: Current oncology (Toronto, Ont.)
• 作者: Solis-Coronado O;Villarreal-Vela MP;Rodríguez-Gutiérrez HF;González-Guerrero JF;Cerda-Flores RM;Alcorta-Núñez F;Camarillo-Cárdenas KP;Pérez-Ibave DC;Vidal-Gutiérrez O;Ramírez-Correa GA;Garza-Rodríguez ML
• 通讯作者: Garza-Rodríguez ML

Prolactin Expression in the Baboon (Papio hamadryas) Eye.

• DOI: 10.3390/ani12172288
• 发表时间: 2022-09-03
• 期刊: Animals : an open access journal from MDPI

Heart Failure-Related Hyperphosphorylation in the Cardiac Troponin I C Terminus Has Divergent Effects on Cardiac Function In Vivo.

• DOI: 10.1161/circheartfailure.117.003850
• 发表时间: 2017-09
• 期刊: Circulation. Heart failure
• 作者: Li Y;Zhu G;Paolocci N;Zhang P;Takahashi C;Okumus N;Heravi A;Keceli G;Ramirez-Correa G;Kass DA;Murphy AM
• 通讯作者: Murphy AM