Protective factors in diabetic kidney disease in patients with type 1 diabetes

1型糖尿病患者糖尿病肾病的保护因素

• 批准号: 10753046
• 负责人: Maria Luiza A Caramori
• 金额: $ 60.67万
• 依托单位: CLEVELAND CLINIC LERNER COM-CWRU
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10753046
• 关键词: Advanced Glycosylation End Products; Area; Attenuated; Base Excision Repairs; Basic Science; Behavior; Biological Assay; Biological Markers; Biological Models; Candidate Disease Gene; Cell Cycle; Cells; Chronic; Collaborations; Complications of Diabetes Mellitus; DNA; DNA Damage; DNA Methylation; DNA Repair; DNA Repair Gene; DNA Repair Pathway; DNA biosynthesis; Data; Deoxyguanosine; Development; Diabetes Mellitus; Diabetic Nephropathy; Electrospray Ionization; End stage renal failure; Epigenetic Process; Epithelial Cells; Excision Repair; Exhibits; Exposure to; Fibroblasts; Functional disorder; Gene Expression; Gene Expression Regulation; Genes; Genetic; Glucose; Histones; Hour; Human; Hyperglycemia; In Vitro; Individual; Inductively Coupled Plasma Mass Spectrometry; Injury; Insulin-Dependent Diabetes Mellitus; Investigation; Knowledge; Laboratories; Lesion; Link; Liquid Chromatography; Measures; Methodology; Methylation; Monozygotic twins; Outcome; Oxidative Stress; Paper; Pathway interactions; Patients; Positioning Attribute; Post-Translational Protein Processing; Process; Proteins; Recovery; Regulation; Renal function; Research; Rodent Model; Role; Skin; Stress; Testing; Therapeutic; Therapeutic Agents; Tubular formation; Twin Multiple Birth; Up-Regulation; Urine; Variant; Work; adduct; cell behavior; cell injury; comparative; comparison control; differential expression; disorder prevention; disorder risk; healing; improved; improved outcome; in vivo; innovation; insight; kidney biopsy; non-diabetic; novel; overexpression; oxidative DNA damage; oxidative damage; prevent; promoter; protective factors; protein expression; pyrosequencing; repaired; response; riboside; tandem mass spectrometry; urinary; volunteer

Scientific Abstract: Diabetic kidney disease (DKD) is the leading cause of end-stage renal disease. While DKD injury factors are extensively studied, protective factors remain poorly defined and present a critical knowledge gap. Recently we and others described the existence of potential protective and healing mechanisms in DKD. These concepts warrant further investigation and have the potential to greatly improve outcomes. Papers from our and other laboratories confirm that after multiple passages cultured skin fibroblasts (SF) behaviors differ between patients with versus without DKD. Consistent with protective mechanisms, our earlier gene expression studies showed that individuals protected from DKD had some SF behaviors that differ both from patients with DKD and from normal controls, suggesting the protected group had unique cellular behaviors associated with very slow development of DKD lesions. Our recent gene expression studies (hi-seq) in SF grown in high glucose (HG) uncovered that pathways related to DNA replication and repair, protein repair, and cell cycle were markedly overexpressed in patients with type 1 diabetes (T1D) without DKD as compared to T1D patients with DKD and non-diabetic controls. These pathways were also differently expressed in SF of monozygotic twins discordant for T1D. Thus, upregulation of these pathways is posited as protective of DKD and epigenetically regulated. Hyperglycemia induces DNA damage, thus activating DNA repair pathways [primarily the base excision repair (BER) pathway] in order to restore DNA integrity and prevent cellular dysfunction. Indeed, the BER pathway was markedly up-regulated in T1D patients without DKD (p=5.14e-5) as compared to patients with DKD and controls. Our preliminary data indicate that BER pathway is also upregulated in proximal tubular epithelial cells (PTEC). We will test whether these pathway differences are associated with functional differences in SF and PTEC cultured from research skin and kidney biopsies in T1D volunteers. Our Specific Aims are to determine 1) changes in BER protein levels, DNA damage and repair capacity in SF in response to in vitro HG and their relationship to DKD risk, 2) changes in BER protein levels, DNA damage, repair capacity and response to a therapeutic agent in PTEC cultured in HG and their relationship to DKD risk, 3) urinary levels of DNA damage products in relation to DKD risk, and 4) epigenetic mechanisms associated with gene expression in key differentially expressed BER pathway genes. We hypothesize that, although both T1D patients with and without DKD may have increased BER pathway activity compared to controls, this activity will be much greater in the patients protected from DKD, and that our robust SF findings will be recapitulated in PTEC. Given the depth of our basic science strengths in the relevant areas, the unique research materials, the innovative technical capabilities of our superb investigative team, and our proven collaboration track, we are extremely well positioned for the successful completion of these studies. If our hypotheses are borne out, this work will open new research avenues aimed at DKD prevention.

科学摘要：糖尿病性肾脏疾病（DKD）是终末期肾脏疾病的主要原因。而DKD 对伤害因素进行了广泛的研究，保护因素的定义较差，并提出了关键知识 差距。最近，我们和其他人描述了DKD中潜在的保护和康复机制的存在。 这些概念需要进一步调查，并有可能大大改善结果。来自 我们和其他实验室证实，在多个段落培养的皮肤成纤维细胞（SF）行为之后 在患有DKD的患者与患者之间。与保护机制一致，我们的早期基因 表达研究表明，受保护不受DKD的个体具有一些SF行为，两者都不同 DKD患者和正常对照患者，表明受保护组具有独特的细胞行为 与DKD病变的发展非常缓慢。我们最近在SF种植的基因表达研究（HI-SEQ） 在高葡萄糖（HG）中，发现与DNA复制和修复有关的途径，蛋白质修复和细胞周期 与T1D患者相比，没有DKD的1型糖尿病（T1D）患者明显过表达 具有DKD和非糖尿病控制。这些途径在单卵双胞胎的SF中也有所不同 与T1D不一致。因此，这些途径的上调被认为是对DKD的保护和表观遗传的 受监管。高血糖诱导DNA损伤，从而激活DNA修复途径[主要是碱基 切除修复（BER）途径]，以恢复DNA完整性并防止细胞功能障碍。确实， 与患有DKD的T1D患者（P = 5.14E-5）相比 DKD和控件。我们的初步数据表明，BER途径也在近端管状中上调 上皮细胞（PTEC）。我们将测试这些途径差异是否与功能差异有关 在T1D志愿者的研究皮肤和肾脏活检中培养的SF和PTEC中。我们的具体目的是 确定1）响应体外HG的BER蛋白水平，DNA损伤和修复能力的变化 以及它们与DKD风险的关系，2）BER蛋白水平，DNA损伤，维修能力和响应的变化 在Hg培养的PTEC中的治疗剂及其与DKD风险的关系，3）DNA损伤水平 与DKD风险有关的产品，以及4）与基因表达相关的表观遗传机制 差异表达的BER途径基因。我们假设这是，尽管有和没有T1D患者 与对照组相比，DKD可能增加了BER途径活动，此活动在 受到DKD保护的患者，我们的鲁棒SF发现将在PTEC中概括。鉴于深度 我们在相关领域，独特的研究材料，创新技术的基础科学优势 我们出色的调查团队的能力以及我们久经考验的合作曲目，我们的位置非常好 为了成功完成这些研究。如果我们的假设得到证实，这项工作将开放新的研究 旨在预防DKD的大道。