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

基本信息

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
项目类别：
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

项目摘要

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：糖尿病肾病（DKD）是终末期肾病的主要原因。而DKD 损伤因素得到了广泛的研究，保护因素仍然不明确，并且提供了关键的知识差距。最近，我们和其他人描述了 DKD 中潜在的保护和治愈机制的存在。这些概念值得进一步研究，并有可能大大改善结果。论文来自我们和其他实验室证实，多次传代后培养的皮肤成纤维细胞 (SF) 的行为有所不同患有 DKD 的患者与未患有 DKD 的患者之间的比较。与保护机制一致，我们早期的基因表达研究表明，免受 DKD 影响的个体具有一些不同于 DKD 的 SF 行为。 DKD 患者和正常对照组的研究结果表明，受保护的群体具有独特的细胞行为与 DKD 病变发展非常缓慢有关。我们最近在 SF 中进行的基因表达研究 (hi-seq) 在高葡萄糖 (HG) 中发现与 DNA 复制和修复、蛋白质修复和细胞周期相关的途径与 T1D 患者相比，在无 DKD 的 1 型糖尿病 (T1D) 患者中显着过度表达具有 DKD 和非糖尿病对照。这些途径在同卵双胞胎的 SF 中也有不同的表达 T1D 不一致。因此，这些途径的上调被认为对 DKD 具有保护作用，并且在表观遗传学上受监管。高血糖会诱导 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 患者相比，BER 通路在无 DKD 的 T1D 患者中显着上调 (p=5.14e-5) DKD 和控制。我们的初步数据表明 BER 通路在近端肾小管中也上调上皮细胞（PTEC）。我们将测试这些途径差异是否与功能差异相关从 T1D 志愿者的皮肤和肾脏活检研究中培养的 SF 和 PTEC。我们的具体目标是确定 1) SF 响应体外 HG 时 BER 蛋白水平、DNA 损伤和修复能力的变化及其与 DKD 风险的关系，2) BER 蛋白水平、DNA 损伤、修复能力和反应的变化 HG 培养的 PTEC 中的治疗剂及其与 DKD 风险的关系，3) 尿液 DNA 损伤水平与 DKD 风险相关的产品，以及 4) 与关键基因表达相关的表观遗传机制差异表达的 BER 通路基因。我们假设，尽管 T1D 患者有或没有与对照组相比，DKD 的 BER 通路活性可能有所增加，这种活性在对照组中会更大。患者免受 DKD 的侵害，并且我们强有力的 SF 研究结果将在 PTEC 中重述。鉴于深度我们在相关领域的基础科学优势、独特的研究材料、创新的技术凭借我们卓越的调查团队的能力以及经过验证的合作轨迹，我们处于非常有利的位置为了顺利完成这些研究。如果我们的假设得到证实，这项工作将开启新的研究旨在预防 DKD 的途径。