Abstract: SA-PO0294
Effects of High Glucose on Tubular Epithelial Cell Gene Expression Cultured at Low Confluency
Session Information
- Diabetic Kidney Disease: Basic and Translational Science Advances - 2
November 08, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Diabetic Kidney Disease
- 701 Diabetic Kidney Disease: Basic
Authors
- Merchant, Michael, University of Louisville School of Medicine, Louisville, Kentucky, United States
- Shartzer, Dimond, University of Kentucky College of Medicine, Lexington, Kentucky, United States
- Barati, Michelle T., University of Louisville School of Medicine, Louisville, Kentucky, United States
Background
Diabetic kidney disease (DKD) detrimentally impacts tubular epithelial cell (TEC) compartment. Maladaptive repair can result in loss of TEC function, TEC drop-out, interstitial fibrosis, and contributes to end-stage renal disease. TEC damage and dropout requires a functional proliferative response to re-establish the TEC monolayer. We conducted studies to address the hypothesis that hyperglycemic growth conditions would regulate TEC gene expression programs as a function of cell confluency.
Methods
Human kidney tubular epithelial cells (HK-11 cell line) were grown to low (<40%) confluency under conditions of normal (5mM) (NG) and high (25mM) glucose (HG) conditions and RNA isolated (n=4/group; 2 groups). Purity/integrity assessed by A260/280 & A260/230 ratios, and Bioanalyzer RIN values. RNA libraries were prepared using the Universal Plus mRNA-Seq with NuQuant, QA/QC assessed using MiSeq Nano Kit V2. Sequenced using Illumina NextSeq 500 with High Output Kit v2.5. Data were analyzed using the informatic pipeline including Fastqc > Trimmomatic > STAR > Cuffnorm/HTSeq > Cuffdiff/DESeq2 > category Compare > cluster profiler (GO and KEGG) to examine differences of the transcriptome and affected pathways. Differences in abundace were determined by ttest (p<0.05) corrected for multiple comparisons (q<0.05).
Results
Differential expression of 331 (p<0.05) transcripts (DEG) in TEC cells were associated with growth in HG vs NG at 40% TEC confluency. 121 DEGs were decreased and 210 DEGS were increased with HG. Six transcirpts were differntially regulated (q<0.05). Two increased in HG and low conflucency (TXNIP and H1FX) and four decreased by HG and low confluency (ZFP36L, CXCL1, BIRC3 and RRAD). Analysis of DEGs by gene ontology indentified roles for protein ubiquitination, apoptotic signaling, nuclear division, hypoxia/unfolded protein response, ER nucleus signaling, and stem cell proliferation.Analysis by KEGG identified significant roles of NF-kappa B and cell cycle.
Conclusion
These data suggest a role for hyperglucoseuria or hyperglucosemia in the TEC reponse of the epithelial cell damage upon low cell confluency. These data suggest TEC differentially regulated genes may be involved in pathways associated with both cell death and or proliferation. These transcripts bear futher investigation as potential routes to understand early DKD damage.
Funding
- NIDDK Support