Target For Diabetic Kidney Disease Research Articles

#71 Dysregulation of ITGAM and ITGB2 in the pathogenesis of diabetic kidney disease—results from bioinformatics analysis and in vitro studies

Abstract Background and Aims Activated neutrophils can release web-like chromatin structures known as neutrophil extracellular traps (NETs), but their roles in the immunopathogenesis of diabetic kidney disease (DKD) remains elusive. Method The characteristic NETs associated targets of differentially expressed genes (DEGs) from human kidney biopsy datasets of patients with DKD (GSE30528 and GSE30529) were identified by machine learning method (LASSO and SVM-RFE). The genes expression was further validated using kidney biopsy dataset of GSE142025. The most abundant single cell of the targets in DKD was further selected by single-nucleus RNA sequencing datasets (GSE131882). Predictive pathway network of targeted genes was constructed using GENEMANIA database. The expressions and potential signaling pathway of these candidate genes were validated using flow cytometry and ELISA. Results Three predicting NETs related genes (ITGAM, ITGB2 and TLR7), identified by machine learning algorithms, which all showed significant upregulation in both glomerular and tubulointerstitial compartments in human DKD kidney samples. Single-cell analysis suggested that ITGAM, ITGB2 and TLR7 were most enriched in leucocytes. DKD patients (n = 18) showed significantly higher activated neutrophils as well as increased expression of ITGAM and ITGB2 compared to healthy controls (HCs, n = 9). DKD patients also exhibited higher serum levels of IL6 but lower IL-10 than HCs (all p < 0.01). The significant network of IL10, IL6, ITGAM and ITGB2 was generated. This network might be associated with leukocyte activation, macrophage activation, JAK-STAT pathway, acute inflammatory response and lymphocyte immune response (all p < 0.05). Conclusion Aberrant ITGAM and ITGB2 expressions in activated neutrophils contribute to DKD pathogenesis and may serve as novel therapeutic targets for DKD.

The Role of Immune Cells in DKD: Mechanisms and Targeted Therapies.

Diabetic kidney disease (DKD), is a common microvascular complication and a major cause of death in patients with diabetes. Disorders of immune cells and immune cytokines can accelerate DKD development of in a number of ways. As the kidney is composed of complex and highly differentiated cells, the interactions among different cell types and immune cells play important regulatory roles in disease development. Here, we summarize the latest research into the molecular mechanisms underlying the interactions among various immune and renal cells in DKD. In addition, we discuss the most recent studies related to single cell technology and bioinformatics analysis in the field of DKD. The aims of our review were to explore immune cells as potential therapeutic targets in DKD and provide some guidance for future clinical treatments.

Potential application of Nardostachyos Radix et Rhizoma-Rhubarb for the treatment of diabetic kidney disease based on network pharmacology and cell culture experimental verification.

Diabetic kidney disease (DKD) is one of the serious complications of diabetes mellitus, and the existing treatments cannot meet the needs of today's patients. Traditional Chinese medicine has been validated for its efficacy in DKD after many years of clinical application. However, the specific mechanism by which it works is still unclear. Elucidating the molecular mechanism of the Nardostachyos Radix et Rhizoma-rhubarb drug pair (NRDP) for the treatment of DKD will provide a new way of thinking for the research and development of new drugs. To investigate the mechanism of the NRDP in DKD by network pharmacology combined with molecular docking, and then verify the initial findings by in vitro experiments. The Traditional Chinese Medicine Systems Pharmacology (TCMSP) database was used to screen active ingredient targets of NRDP. Targets for DKD were obtained based on the Genecards, OMIM, and TTD databases. The VENNY 2.1 database was used to obtain DKD and NRDP intersection targets and their Venn diagram, and Cytoscape 3.9.0 was used to build a "drug-component-target-disease" network. The String database was used to construct protein interaction networks. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis and Gene Ontology analysis were performed based on the DAVID database. After selecting the targets and the active ingredients, Autodock software was used to perform molecular docking. In experimental validation using renal tubular epithelial cells (TCMK-1), we used the Cell Counting Kit-8 assay to detect the effect of NRDP on cell viability, with glucose solution used to mimic a hyperglycemic environment. Flow cytometry was used to detect the cell cycle progression and apoptosis. Western blot was used to detect the protein expression of STAT3, p-STAT3, BAX, BCL-2, Caspase9, and Caspase3. A total of 10 active ingredients and 85 targets with 111 disease-related signaling pathways were obtained for NRDP. Enrichment analysis of KEGG pathways was performed to determine advanced glycation end products (AGEs)-receptor for AGEs (RAGE) signaling as the core pathway. Molecular docking showed good binding between each active ingredient and its core targets. In vitro experiments showed that NRDP inhibited the viability of TCMK-1 cells, blocked cell cycle progression in the G0/G1 phase, and reduced apoptosis in a concentration-dependent manner. Based on the results of Western blot analysis, NRDP differentially downregulated p-STAT3, BAX, Caspase3, and Caspase9 protein levels (P < 0.01 or P < 0.05). In addition, BAX/BCL-2 and p-STAT3/STAT3 ratios were reduced, while BCL-2 and STAT3 protein expression was upregulated (P < 0.01). NRDP may upregulate BCL-2 and STAT3 protein expression, and downregulate BAX, Caspase3, and Caspase9 protein expression, thus activating the AGE-RAGE signaling pathway, inhibiting the vitality of TCMK-1 cells, reducing their apoptosis. and arresting them in the G0/G1 phase to protect them from damage by high glucose.

Therapeutic Targets for Diabetic Kidney Disease: Proteome-Wide Mendelian Randomization and Colocalization Analyses.

At present, safe and effective treatment drugs are urgently needed for diabetic kidney disease (DKD). Circulating protein biomarkers with causal genetic evidence represent promising drug targets, which provides an opportunity to identify new therapeutic targets. Summary data from two protein quantitative trait loci (pQTL) studies: one involving 4,907 plasma proteins data from 35,559 individuals, and the other encompassing 4,657 plasma proteins among 7,213 European Americans. Summary statistics for DKD were obtained from a large genome-wide association study (3345 cases and 2372 controls) and the FinnGen study (3676 cases and 283,456 controls). Mendelian randomization (MR) analysis was conducted to examine the potential targets for DKD. The colocalization analysis was utilized to detect whether the potential proteins exist the shared causal variants. To enhance the credibility of the results, external validation was conducted. Additionally, enrichment analysis, assessment of protein druggability, and the protein-protein interaction (PPI) networks were employed to further enrich the research findings. The proteome-wide MR analyses identified 21 blood proteins that may causally be associated with DKD. Colocalization analysis further supported a causal relationship between 12 proteins and DKD, with external validation confirming four of these proteins, and TGFBI was affirmed through two separate group datasets. These results indicate that targeting these four proteins could be a promising approach for treating DKD, and warrant further clinical investigations.

(Pro)renin receptor mediates tubular epithelial cell pyroptosis in diabetic kidney disease via DPP4-JNK pathway

Background(Pro)renin receptor (PRR) is highly expressed in renal tubules, which is involved in physiological and pathological processes. However, the role of PRR, expressed in renal tubular epithelial cells, in diabetic kidney disease (DKD) remain largely unknown.MethodsIn this study, kidney biopsies, urine samples, and public RNA-seq data from DKD patients were used to assess PRR expression and cell pyroptosis in tubular epithelial cells. The regulation of tubular epithelial cell pyroptosis by PRR was investigated by in situ renal injection of adeno-associated virus9 (AAV9)-shRNA into db/db mice, and knockdown or overexpression of PRR in HK-2 cells. To reveal the underlined mechanism, the interaction of PRR with potential binding proteins was explored by using BioGrid database. Furthermore, the direct binding of PRR to dipeptidyl peptidase 4 (DPP4), a pleiotropic serine peptidase which increases blood glucose by degrading incretins under diabetic conditions, was confirmed by co-immunoprecipitation assay and immunostaining.ResultsHigher expression of PRR was found in renal tubules and positively correlated with kidney injuries of DKD patients, in parallel with tubular epithelial cells pyroptosis. Knockdown of PRR in kidneys significantly blunted db/db mice to kidney injury by alleviating renal tubular epithelial cells pyroptosis and the resultant interstitial inflammation. Moreover, silencing of PRR blocked high glucose-induced HK-2 pyroptosis, whereas overexpression of PRR enhanced pyroptotic cell death of HK-2 cells. Mechanistically, PRR selectively bound to cysteine-enrich region of C-terminal of DPP4 and augmented the protein abundance of DPP4, leading to the downstream activation of JNK signaling and suppression of SIRT3 signaling and FGFR1 signaling, and then subsequently mediated pyroptotic cell death.ConclusionsThis study identified the significant role of PRR in the pathogenesis of DKD; specifically, PRR promoted tubular epithelial cell pyroptosis via DPP4 mediated signaling, highlighting that PRR could be a promising therapeutic target in DKD.

Exercise alleviates diabetic kidney disease through PPAR��-CPT1�� pathway-dependent fatty acid ��-oxidation

&lt;p&gt;Diabetic kidney disease (DKD), a severe diabetic complication affecting approximately one-third of diabetic patients, is the leading cause of end-stage chronic kidney disease. The benefits of regular exercise for patients with DKD have been well documented, particularly in overweight patients with DKD. However, the underlying mechanisms are incompletely understood. The present study demonstrates that exercise improves kidney function in diabetic &lt;i&gt;db/db&lt;/i&gt; mice through activating PPAR��-mediated fatty acid ��-oxidation (FAO). Twelve-week treadmill running exercise improved kidney function in &lt;i&gt;db/db&lt;/i&gt; mice. Metabolomics and transcriptomics profiling analysis collectively revealed that dysregulation of FAO in &lt;i&gt;db/db&lt;/i&gt; mice was largely corrected by running exercise. KEGG pathway enrichment revealed that PPAR pathway, a critical signalling cascade in FAO and exercise, is involved in DKD, suggesting PPAR�� activation protects renal function through promoting FAO. Which was confirmed by improved renal function and enhanced FAO in &lt;i&gt;db/db&lt;/i&gt; mice treated with PPAR�� agonist GW501516. Oppositely, PPAR��-specific inhibitor GSK0660 abolished exercise-mediated improvement in renal function and FAO. In combining with gene expression profiling data we identified CPT1��, the rate-limiting enzyme in fatty acid oxidation, was suppressed in the kidney of diabetic mice and reversed by exercise and the PPAR�� agonist GW501516. The findings hint at a potential role for the PPAR��-CPT1�� pathway in exercise-induced improvements in diabetic renal function and may warrant further exploration of the PPAR��-CPT1�� pathway as a therapeutic target in DKD.&lt;/p&gt;

Insights Into the Functional Mechanism of Diabetic Kidney Disease Treatment with Sinensetin Based on Network Pharmacology and Molecular Docking

ABSTRACT Objective: Sinensetin is an antioxidant widely used for treating various diseases; however, its function and targets in diabetic kidney disease (DKD) remain unknown. This study aimed to investigate the functional mechanism of sinensetin in DKD using network pharmacological and molecular docking methods. Methods: Potential targets of sinensetin for DKD treatment were identified using the Swiss Target Prediction (http://www.swisstargetprediction.ch/), STITCH (http://stitch.embl.de/), Online Mendelian Inheritance in Man (OMIM, https://omim.org/), and GeneCards (https://www.genecards.org/) databases. These potential targets were introduced into the STRING database to analyze protein–protein interactions (PPI) and identify key targets. These key targets were subjected to Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The binding activities of these key targets were confirmed using molecular docking analysis. Results: A total of 53 key targets of sinensetin treatment in DKD were identified. Ten core targets, including protein kinase SRC (SRC), epidermal growth factor receptor (EGFR), and prostaglandin-endoperoxide synthase 2 (PTGS2) were identified using PPI network interactions. GO functional analysis revealed that reactive oxygen species metabolic processes, membrane rafts, and protein tyrosine kinase activity were involved in sinensetin treatment of DKD. The KEGG enrichment analysis exhibited that signaling pathways such as pathways in cancer, tryptophan metabolism, and the vascular endothelial growth factor (VEGF) signaling pathway play important roles. Molecular docking analysis suggested favorable docking between the ten core targets and sinensetin. Conclusion: Sinensetin plays a role in the treatment of DKD through various biological processes and signaling pathways. These data provide theoretical evidence for experimental studies on DKD treatment using sinensetin.

Integrin α3β1 as a Potential Target for Diabetic Kidney Disease (DKD)

Integrin α3β1 as a Potential Target for Diabetic Kidney Disease (DKD)

Exosomal microRNAs: potential nanotherapeutic targets for diabetic kidney disease.

Diabetic kidney disease (DKD) is a primary cause for end-stage renal disease, but no specific therapeutic approaches exist. Exosomal miRNAs, a key functional cargo of nanovesicles, play crucial roles in the pathophysiological processes of DKD. Exosomal miRNAs are involved in cell-to-cell transfer of biological information, mediating nephritic inflammation, oxidative stress, apoptosis, autophagy, epithelial-mesenchymal transition and fibrosis. Circulating exosomal miRNAs derived from urine or serum might function as noninvasive prognostic biomarkers for DKD. Exosomal miRNAs from stem cells have been reported to exert beneficial effects on diabetic kidneys, which suggests that these exosomes might function as potential nanotherapy tools for treating DKD. In this review, we have summarized recent studies based on the association between exosomal miRNAs and DKD.

Key enzyme in charge of ketone reabsorption of renal tubular SMCT1 may be a new target in diabetic kidney disease.

A ketogenic diet or mildly increased ketone body levels are beneficial for diabetic kidney disease (DKD) patients. Our previous study has found that sodium-coupled monocarboxylate transporter 1 (SMCT1), a key enzyme in charge of ketone reabsorption, possesses beneficial effects on the function of renal tubular epithelial cells (TECs) in energy crisis. Our present study is to investigate whether SMCT1 is important in maintaining the physiological function of renal tubular and plays a role in DKD. We tested the expression of SMCT1 in kidney tissues from DKD patients receiving kidney biopsy as well as diabetes mice. We compared the difference of β-hydroxybutyrate (β-HB) levels in serum, urine and kidney tissues between diabetic mice and control. Using recombinant adeno-associated viral vector containing SMCT1 (encoded by Slc5a8 gene), we tested the effect of SMCT1 upregulation on microalbuminuria as well as its effects on mitochondrial energy metabolism in diabetic mice. Then we investigated the role of SMCT1 and its β-HB reabsorption function in maintaining the physiological function of renal tubular using renal tubule-specific Slc5a8 gene knockout mice. Transcriptomes and proteomics analysis were used to explore the underlying mechanism. SMCT1 downregulation was found in DKD patients as well as in diabetic mice. Moreover, diabetic mice had a decreased renal β-HB level compared with control, and SMCT1 upregulation could improve microalbuminuria and mitochondrial energy metabolism. In renal tubule-specific Slc5a8 gene knockout mice, microalbuminuria occurred early at 24 weeks of age, accompanied by ATP shortage and metabolic reprogramming in the kidney; however, supplementation with β-HB precursor substance 1,3-butanediol in food alleviated kidney damage as well as energy metabolic reprogramming. Decreased SMCT1 expression and its ketone reabsorption function play an important role in the occurrence of DKD. SMCT1 may be a new promising target in treating DKD.

10-LB: Activation of Sigma-1 Receptor as a Novel Therapeutic Target in Diabetic Kidney Disease

Introduction and aims: Diabetic kidney disease (DKD) is a major health concern among diabetic patients. Current therapies cannot halt the progression of renal failure; thus, the development of additional novel therapies is of paramount importance. We previously showed that the Sigma-1 receptor (S1R) agonism is protective in acute kidney injury (AKI). Inflammation and tubular hypoxia are key processes in the pathogenesis of DKD. S1R agonist treatment inhibits these pathologic pathways in AKI, therefore we assume that a similar protective effect can be shown in DKD as well. Methods: Diabetes mellitus (DM) was induced by streptozotocin (65 mg/bwkg, ip.) in adult, male Wistar rats and they were treated for seven weeks with fluvoxamine (D+FLU, 20 mg/bwkg/day, po.). Metabolic parameters, renal function, and histology were evaluated. Inflammation was tested in lipopolysaccharide-stimulated (LPS, 1 µg/mL) human proximal tubular epithelial cells (HK-2). To test hypoxia cells were placed in a hypoxic chamber (1% O2, 2h). Cells were treated with 10 µM FLU. Proinflammatory and hypoxia markers were measured. Results: Renal function (BUN: DM: 26.6±2.42 vs. DM+FLU: 17.3±2.30 mmol/L; Serum creatinine: DM: 42.0±2.39 vs.DM+FLU: 34.5±2.74 µmol/L; Proteinuria: DM: 42.5±6.38 vs. DM+FLU: 21.5±4.99 mg/mL; p&amp;lt;0.05), structural damage (PAS staining), and fibrotic tissue accumulation (Masson’s trichrome staining) were reduced by FLU. FLU decreased LPS-induced IL6, TLR4, and NFKB1 expressions in HK-2 cells. Hypoxia-induced HIF1A and HIF2A elevation were suspended by FLU treatment. Moreover, VEGFA and SLC2A1 expressions were also increased in hypoxia and FLU prevented their elevation. Conclusions: Activation of S1R slows the progression of chronic renal impairment in DKD. Moreover, S1R agonist FLU prevents hypoxic and inflammatory damage in HK-2 cells, which may explain the molecular mechanisms behind its renoprotective effect. We propose S1R as a novel target for the prevention of DKD. Disclosure D. B. Balogh: None. L. Lenart: None. A. Hosszu: None. A. Saeed: None. J. Hodrea: None. A. R. Toth: None. A. J. Szabo: None. A. Fekete: None. Funding National Research, Development and Innovation Fund (UNKP-22-4-II-SE-2, TKP2021-EGA-24); Hungarian Academy of Sciences (LP2021-3/2021)

Exploring the pharmacological mechanisms of Tripterygium wilfordii against diabetic kidney disease using network pharmacology and molecular docking

BackgroundTripterygium wilfordii (TW), when formulated in traditional Chinese medicine (TCM), can effectively treat diabetic kidney disease (DKD). However, the pharmacological mechanism associated with its success has not yet been elucidated. The current work adopted network pharmacology and molecular docking for exploring TW-related mechanisms in treating DKD. Methods: In the present work, the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database was employed to obtain the effective components and candidate targets of TW. Additionally, this work utilized the UniProt protein database for screening and standardizing human-derived targets for effective components. The Cytoscape software was utilized to construct an effective component-target network for TW. Targets for DKD were acquired in the GEO, DisGeNET, GeneCards, and OMIM databases. Additionally, a Venn diagram was also plotted to select the possible targets of TW for treating DKD. Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted to explore the TW-related mechanism underlying DKD treatment. This work also built a protein-protein interaction (PPI) network based on the Cytoscape and String platform. Then, molecular docking was conducted in order to assess the affinity of key proteins for related compounds. Results: In total, 29 active components and 134 targets of TW were acquired, including 63 shared targets, which were identified as candidate therapeutic targets. Some key targets and important pathways were included in the effect of TW in treating DKD. Genes with higher degrees, including TNF and AKT1, were identified as hub genes of TW against DKD. Molecular docking showed that TNF and AKT1 bind well to the main components in TW (kaempferol, beta-sitosterol, triptolide, nobiletin, and stigmasterol). ConclusionsTW primarily treats DKD by acting on two targets (AKT1 and TNF) via the five active ingredients kaempferol, beta-sitosterol, triptolide, nobiletin, and stigmasterol.

Identification and validation of immune-related biomarkers and potential regulators and therapeutic targets for diabetic kidney disease

BackgroundDiabetic kidney disease (DKD) is a major complication of diabetes and the leading cause of end-stage renal disease worldwide. Renal inflammation and infiltration of immune cells contribute to the development and progression of DKD. Thus, the aim of the present study was to identify and validate immune-related biomarkers and analyze potential regulators including transcription factors (TFs), microRNAs (miRNAs), and drugs for DKD.MethodsImmune-related genes from the ImmPort database and glomeruli samples from GSE1009 and GSE30528 were used to identify differentially expressed immune-related genes (DEIRGs) of DKD. The expression level and clinical correlation analyses of DEIRGs were verified in the Nephroseq database. Murine podocytes were cultured to construct the high glucose-induced podocyte injury model. The reliability of the bioinformatics analysis was experimentally validated by RT-qPCR in podocytes. Networks among DEIRGs, regulators, and drugs were constructed to predict potential regulatory mechanisms for DKD.ResultsDKD-associated DEIRGs were identified. CCL19 and IL7R were significantly upregulated in the DKD group and negatively correlated with glomerular filtration rate (GFR). GHR, FGF1, FYN, VEGFA, F2R, TGFBR3, PTGDS, FGF9, and SEMA5A were significantly decreased in the DKD group and positively correlated with GFR. RT-qPCR showed that the relative mRNA expression levels of GHR, FGF1, FYN, TGFBR3, PTGDS, FGF9, and SEMA5A were significantly down-regulated in the high glucose-induced podocyte injury group. The enriched regulators for DEIRGs included 110 miRNAs and 8 TFs. The abnormal expression of DEIRGs could be regulated by 16 established drugs.ConclusionsThis study identified immune-related biomarkers, regulators, and drugs of DKD. The findings of the present study provide novel insights into immune-related diagnosis and treatment of DKD.

Hyaluronan Inhibition as a Therapeutic Target for Diabetic Kidney Disease: What Is Next?

Diabetic kidney disease (DKD) is the leading cause of CKD and ESKD in the United States and worldwide. Pharmacotherapy and lifestyle modifications for glycemia, dyslipidemia, and BP control have shown success in slowing the progression of DKD. Traditional treatments, such as angiotensin-converting enzyme inhibitors or angiotensin receptor blockers and more recently the use of sodium-glucose cotransporter 2 inhibitors, nonsteroidal selective mineralocorticoid receptor antagonists, such as finerenone, and glucagon-like peptide 1 receptor agonists, have led to added benefits on various outcomes. However, significant residual risk for DKD progression remains despite the current standard-of-care approaches. Arteriolar hyalinosis (AH) is among the key findings seen on kidney biopsies of patients with DKD. It results from the excessive accumulation of hyaluronan (HA) in the arterioles. AH has not been targeted specifically by any of the therapeutic methods currently being used. We discuss in this manuscript the potential use of a selective therapy targeting AH and the increased total renal HA deposits using a HA synthesis inhibitor in DKD.

Rho-kinase inhibitor restores glomerular fatty acid metabolism in diabetic kidney disease

The small GTPase Rho and its effector Rho-kinase (ROCK) are activated in the diabetic kidney, and recent studies decade have demonstrated that ROCK signaling is an integral pathway in the progression of diabetic kidney disease. We previously identified the distinct role of ROCK1, an isoform of ROCK, in fatty acid metabolism in diabetic glomeruli. However, the effect of pharmacological intervention for ROCK1 is not clear. In the present study, we show that the inhibition of ROCK1 by Y-27632 and fasudil restores fatty acid oxidation in the glomeruli. Mechanistically, these compounds optimize fatty acid utilization and redox balance in mesangial cells via AMPK phosphorylation and the subsequent induction of PGC-1α. A further in vivo study showed that the inhibition of ROCK1 suppressed the downregulation of the fatty acid oxidation-related gene expression in glomeruli and mitochondrial fragmentation in the mesangial cells of db/db mice. These observations indicate that ROCK1 could be a promising therapeutic target for diabetic kidney disease through a mechanism that improves glomerular fatty acid metabolism.

A Comprehensive Weighted Gene Co-expression Network Analysis Uncovers Potential Targets in Diabetic Kidney Disease.

Diabetic kidney disease (DKD) is one of the most common microvascular complications of diabetes. It has always been difficult to explore novel biomarkers and therapeutic targets of DKD. We aimed to identify new biomarkers and further explore their functions in DKD. The weighted gene co-expression network analysis (WGCNA) method was used to analyze the expression profile data of DKD, obtain key modules related to the clinical traits of DKD, and perform gene enrichment analysis. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the mRNA expression of the hub genes in DKD. Spearman's correlation coefficients were used to determine the relationship between gene expression and clinical indicators. Fifteen gene modules were obtained via WGCNA analysis, among which the green module had the most significant correlation with DKD. Gene enrichment analysis revealed that the genes in this module were mainly involved in sugar and lipid metabolism, regulation of small guanosine triphosphatase (GTPase) mediated signal transduction, G protein-coupled receptor signaling pathway, peroxisome proliferator-activated receptor (PPAR) molecular signaling pathway, Rho protein signal transduction, and oxidoreductase activity. The qRT-PCR results showed that the relative expression of nuclear pore complex-interacting protein family member A2 (NPIPA2) and ankyrin repeat domain 36 (ANKRD36) was notably increased in DKD compared to the control. NPIPA2 was positively correlated with the urine albumin/creatinine ratio (ACR) and serum creatinine (Scr) but negatively correlated with albumin (ALB) and hemoglobin (Hb) levels. ANKRD36 was positively correlated with the triglyceride (TG) level and white blood cell (WBC) count. NPIPA2 expression is closely related to the disease condition of DKD, whereas ANKRD36 may be involved in the progression of DKD through lipid metabolism and inflammation, providing an experimental basis to further explore the pathogenesis of DKD.

A comprehensive weighted gene co-expression network analysis uncovers potential targets in diabetic kidney disease

Abstract Background and Objectives Diabetic kidney disease (DKD) is one of the most common microvascular complications of diabetes. It has always been difficult to explore novel biomarkers and therapeutic targets of DKD. We aimed to identify new biomarkers and further explore their functions in DKD. Methods The weighted gene co-expression network analysis (WGCNA) method was used to analyze the expression profile data of DKD, obtain key modules related to the clinical traits of DKD, and perform gene enrichment analysis. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the mRNA expression of the hub genes in DKD. Spearman’s correlation coefficients were used to determine the relationship between gene expression and clinical indicators. Results Fifteen gene modules were obtained via WGCNA analysis, among which the green module had the most significant correlation with DKD. Gene enrichment analysis revealed that the genes in this module were mainly involved in sugar and lipid metabolism, regulation of small guanosine triphosphatase (GTPase) mediated signal transduction, G protein-coupled receptor signaling pathway, peroxisome proliferator-activated receptor (PPAR) molecular signaling pathway, Rho protein signal transduction, and oxidoreductase activity. The qRT-PCR results showed that the relative expression of nuclear pore complex-interacting protein family member A2 (NPIPA2) and ankyrin repeat domain 36 (ANKRD36) was notably increased in DKD compared to the control. NPIPA2 was positively correlated with the urine albumin/creatinine ratio (ACR) and serum creatinine (Scr) but negatively correlated with albumin (ALB) and hemoglobin (Hb) levels. ANKRD36 was positively correlated with the triglyceride (TG) level and white blood cell (WBC) count. Conclusion NPIPA2 expression is closely related to the disease condition of DKD, whereas ANKRD36 may be involved in the progression of DKD through lipid metabolism and inflammation, providing an experimental basis to further explore the pathogenesis of DKD.

ODP200 Fyn is involved in the development of diabetic kidney disease by modulating autophagy

Abstract Diabetic kidney disease (DKD) is one of the major diabetic complications and the leading cause of the end stage renal disease. Recently autophagy was shown to regulate DKD. Previously we reported that Fyn regulates muscle mass by suppressing autophagy through Fyn-STAT3 signaling pathway. More recently, we demonstrated that Fyn also inhibits autophagy in HK2 cells, in vitro cell model of renal proximal tubular epithelial cells (RPTC). Moreover, we found Y369 of Tgm2, a known inhibitor of autophagy, is directory phosphorylated by Fyn both in vitro and in cells and Fyn-dependent phosphorylation of Tgm2 regulates autophagy in RPTC. It has been reported that Tgm2 forms complexes with p53 and p62 (a known autophagy regulator) to mediate the degradation of p53 at autophagosomes in cancer cells. To note is that p53 could function as a DKD inducer. We found that p53 expression was decreased in Tgm2 knock-downed HK2 cells, suggesting that Tgm2-p62-p53 complex also modulates autophagy in RPTC. Since these were only in vitro studies, we proceeded with those in vivo. In HFD fed mice, diet-induced rodent models of metabolic disorders, we found that protein expression of p53 was increased due to decreased levels of autophagy implicated by decreased p62 punctuations in RPTC. To confirm the role of Fyn in autophagy and p53 expression we took advantage of a diabetic mouse model of FynKO mice induced by Streptozocin(STZ). While p53 was increased in the kidney of diabetic WT mice induced by STZ, it was not the case in the kidney of diabetic FynKO, indicating that Fyn could regulate autophagic activity and p53 expression. Taken together, these data suggest that the metabolic status may regulate Fyn to phosphorylate Tgm2 and modulate Tgm2-p62-p53 complex leading to the development of DKD. This regulatory mechanism is not mediated by blood glucose and may lead to the new therapeutic targets for diabetic kidney disease regardless of control of diabetes. Presentation: No date and time listed

Podocyte as the link between sterile inflammation and diabetic kidney disease

Shahzad etal. examined the underlying mechanisms of sterile inflammation in diabetic kidney disease, specifically the role of NLRP3 inflammasome activation in podocytes. Using mouse models with gain-of-function and loss-of-function mutations in podocyte Nlrp3, or caspase-1 loss-of-function mutations in podocytes, they identified that Nlrp3 activation in these cells is central for development of diabetic kidney disease but not solely dependent on canonical mechanisms and caspase-1. These findings position podocyte-mediated immune cell-like functions as potential therapeutic targets for diabetic kidney disease.

Metabolic reprogramming: A novel therapeutic target in diabetic kidney disease.

Diabetic kidney disease (DKD) is one of the most common microvascular complications of diabetes mellitus. However, the pathological mechanisms contributing to DKD are multifactorial and poorly understood. Diabetes is characterized by metabolic disorders that can bring about a series of changes in energy metabolism. As the most energy-consuming organs secondary only to the heart, the kidneys must maintain energy homeostasis. Aberrations in energy metabolism can lead to cellular dysfunction or even death. Metabolic reprogramming, a shift from mitochondrial oxidative phosphorylation to glycolysis and its side branches, is thought to play a critical role in the development and progression of DKD. This review focuses on the current knowledge about metabolic reprogramming and the role it plays in DKD development. The underlying etiologies, pathological damages in the involved cells, and potential molecular regulators of metabolic alterations are also discussed. Understanding the role of metabolic reprogramming in DKD may provide novel therapeutic approaches to delay its progression to end-stage renal disease.