Epigenetic Control of Kidney Fibrosis

肾脏纤维化的表观遗传控制

基本信息

批准号：
9067142
负责人：
WENZHENG ZHANG
金额：
$ 23.7万
依托单位：
ALBANY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-03-01 至 2020-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9067142
关键词：
Ablation Adverse effects Age Albuminuria Aldosterone Atrasentan Biological Models Biopsy Blood Cardiac Cells Cessation of life Chronic Kidney Failure Cicatrix Clinical Research DNA Data Development Diabetes Mellitus Diabetic Nephropathy Disease Disease Progression Duct (organ) structure Edema End stage renal failure Endothelin-1 Epigenetic Process Fibrosis Foundations Gene Targeting Genes Genetic Genetic Markers Genetic Screening Genetic screening method Glomerular Filtration Rate Goals HK2 gene Health Histone H3 Human Hypertension Hypotension Infusion procedures Injury Kidney Kidney Failure Knock-out Knockout Mice Link Malignant Neoplasms Mediating Methyltransferase Modeling Molecular Mus Mutation Patients Pharmaceutical Preparations Physiological Play Polycystic Kidney Diseases Pre-Clinical Model Public Health Publishing Renal function Renin-Angiotensin-Aldosterone System Repression Role Site Somatic Mutation Spironolactone Streptozocin Susceptibility Gene Technology Testing Tubular formation Up-Regulation Vasoconstrictor Agents Work autocrine cost diagnostic biomarker effective therapy functional loss genetic manipulation high risk hyperkalemia improved in vivo inhibitor/antagonist laser capture microdissection mouse model new therapeutic target next generation sequencing novel novel therapeutics paracrine preconditioning targeted treatment

项目摘要

DESCRIPTION (provided by applicant): Kidney fibrosis is the hallmark of chronic kidney disease (CKD). Despite aggressive management, CKD often progresses to end-stage renal disease, which costs the US >$40 billion dollars and >90,000 deaths annually. The current main therapy targeting the renin-angiotensin-aldosterone (aldo) system with drugs including Spironolactone often delays, but does not stop the progression. This is also true for all other drugs such as endothelin 1 (ET1) blocker Atrasentan. The ineffectiveness and side effects including hyperkalemia and edema necessitate identification of novel therapeutic targets for the development of more effective treatments. Factors modulating the aldo global effect from its primary action site connecting tubule/collecting duct (CNT/CD) may prove better targets. However, such genetic and epigenetic factors remain virtually unknown, partially because of the intrinsic limitations of the clinical studies. These limitations include lack of kidney biopsies to verify the status of the disease, impossibility of genetic manipulation in patients to establish th causative relationship, and impracticability through mutational analyses with blood DNA to identify somatic mutations, which occur at atypical high rate in human kidney. Our published and preliminary data suggest that 1) Patients with diabetic nephropathy (DN) and CKD may have mutations in histone H3 K79 methyltransferase hDOT1L and abolished H3 dimethylation (H3m2K79) in their kidney biopsies; 2) Dot1a (encoded by Dot1l) represses ET1 and other aldo target genes. Aldo relieves Dot1a-mediated repression by multiple mechanisms; 3) CNT/CD-specific ablation of Dot1l in Dot1lAC mice causes abolition of H3m2K79, upregulation of ET1, and development of severe kidney fibrosis throughout the whole kidney. Accordingly, in this proposal, we will develop genetic markers to overcome the above limitations. To this end, we will use kidney biopsies from patients with DN and CKD, our mouse models bearing intact or disrupted Dot1l and ET1 in the CNT/CD, and their CNT/CD primary cells in combination of cutting-edge technologies including laser capture microdissection, next generation sequencing, and in vivo lineage tracing. Our specific aims are to study if DN and CKD patients have genetic defects in hDOT1L (Aim 1), study if Dot1l deletion accelerates kidney fibrosis in part by upregulating ET1 in mice (Aim 2), and study if Dot1a and ET1 modulate the global effect of aldo profibrotic action (Aim 3). Our studies may identify DOT1L as a novel repressor of ET1 and thus a new renoprotective factor, confirm loss of DOT1L function and thus H3m2K79 as an epigenetic driver of CKD, define Spironolactone + Atrasentan as a new effective combinational therapy of CKD, and lay the foundation of new genetic tests. If Dot1l and ET1 are genetically linked to CKD in humans, they may be exploited to develop genetic screening tests to identify patients at high risk of CKD and to determine their responsiveness to various aldo and ET1 inhibitors. Like ET1, DOT1L can also be considered as a potential new therapeutic target of CKD.

描述（由申请人提供）：肾纤维化是慢性肾脏疾病（CKD）的标志。尽管有积极的管理，但CKD经常发展为终末期肾脏疾病，每年损失400亿美元和> 90,000人死亡。目前针对肾素 - 血管紧张素 - 醛固酮（ALDO）系统的主要疗法，包括螺内酯在内的药物通常会延迟，但不会停止进展。对于所有其他药物，例如内皮素1（ET1）阻滞剂Atrasentan也是如此。包括高钾血症和水肿在内的无效性和副作用需要鉴定出新的治疗靶标，以开发更有效的治疗方法。从其主要动作部位连接小管/收集管（CNT/CD）调节ALDO全球效应的因素可能是更好的目标。然而，这种遗传和表观遗传因素几乎是未知的，部分原因是临床研究的内在局限性。这些限制包括缺乏肾脏活检验证疾病的状态，患者的遗传操作不可能建立病因关系，以及通过使用血液DNA进行突变分析以鉴定体细胞突变，这在人肾脏中以非典型高率发生。我们已发表的初步数据表明，1）患有糖尿病性肾病（DN）和CKD的患者在组蛋白H3 K79甲基转移酶HDOT1L中可能具有突变，并废除了H3二甲基化（H3M2K79）的肾脏活检中的H3二甲基化（H3M2K79）； 2）dot1a（由dot1l编码）抑制ET1和其他aldo靶基因。 Aldo通过多种机制缓解了DOT1A介导的抑制作用； 3）DOT1LAC小鼠中DOT1L的CNT/CNT/CD特异性消融会导致H3M2K79的废除，ET1的上调以及整个肾脏中严重的肾纤维化的发展。因此，在此提案中，我们将开发遗传标记以克服上述局限性。为此，我们将使用来自DN和CKD患者的肾脏活检，在CNT/CD中带有完整或破坏DOT1L和ET1的小鼠模型，以及其CNT/CD主细胞与尖端技术的结合，包括激光捕获微分辨率，下一代测序以及InvivoEneage traceage Tracing。我们的具体目的是研究DN和CKD患者在HDOT1L中是否患有遗传缺陷（AIM 1），研究DOT1L缺失是否会加速肾脏纤维化，部分原因是在小鼠中上调ET1（AIM 2），并研究DOT1A和ET1是否调节Aldo纤维化动作的全球效应（AIM AIM 3）。我们的研究可能将DOT1L识别为ET1的新型阻遏物，因此是一种新的肾脏保护因子，确认DOT1L功能的丧失，因此H3M2K79是CKD的表观遗传驱动器，将螺旋罗替酮 + Atrasentan定义为CKD的新有效组合治疗，并为新的遗传试验提供了基础。如果DOT1L和ET1与人类中的CKD遗传有关，则可以利用它们来开发遗传筛查测试，以鉴定患有CKD高风险的患者，并确定他们对各种Aldo和ET1抑制剂的反应性。像ET1一样，DOT1L也可以被视为CKD的潜在新治疗靶标。

项目成果

期刊论文数量（23）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Dot1l deficiency leads to increased intercalated cells and upregulation of V-ATPase B1 in mice.

DOI：
10.1016/j.yexcr.2015.09.014
发表时间：
2016-06-10
期刊：
Experimental cell research
影响因子：
3.7
作者：
Xiao Z;Chen L;Zhou Q;Zhang W
通讯作者：
Zhang W

AF17 facilitates Dot1a nuclear export and upregulates ENaC-mediated Na+ transport in renal collecting duct cells.

DOI：
10.1371/journal.pone.0027429
发表时间：
2011
期刊：
PloS one
影响因子：
3.7
作者：
Wu H;Chen L;Zhou Q;Zhang W
通讯作者：
Zhang W

Widely expressed Af17 is likely not required for embryogenesis, hematopoiesis, and animal survival.

DOI：
10.1002/dvg.20679
发表时间：
2010-12
期刊：
GENESIS
影响因子：
1.5
作者：
Zhang, Zhijing;Huang, Le;Reisenauer, Mary Rose;Wu, Hongyu;Chen, Lihe;Zhang, Yujin;Xia, Yang;Zhang, Wenzheng
通讯作者：
Zhang, Wenzheng

Insights into cellular and molecular basis for urinary tract infection in autosomal-dominant polycystic kidney disease.

深入了解常染色体显性多囊肾病尿路感染的细胞和分子基础。

DOI：
10.1152/ajprenal.00279.2017
发表时间：
2017
期刊：
American journal of physiology. Renal physiology
影响因子：
0
作者：
Gao,Chao;Zhang,Long;Zhang,Ye;Wallace,DarrenP;Lopez-Soler,ReynoldI;Higgins,PaulJ;Zhang,Wenzheng
通讯作者：
Zhang,Wenzheng

Epigenetics of epithelial Na(+) channel-dependent sodium uptake and blood pressure regulation.

上皮Na(+)通道依赖性钠摄取和血压调节的表观遗传学。