Abstract: FR-PO131
The Human MicroRNA-mRNA Transcriptome in AKI
Session Information
- AKI: Mechanisms - II
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- De Luca, Thomas, Indiana University School of Medicine, Indianapolis, Indiana, United States
- Melo ferreira, Ricardo, Indiana University School of Medicine, Indianapolis, Indiana, United States
- Eadon, Michael T., Indiana University School of Medicine, Indianapolis, Indiana, United States
- Janosevic, Danielle, Indiana University School of Medicine, Indianapolis, Indiana, United States
Background
MicroRNAs (miR) regulate gene expression by base-pairing to messenger RNA (RNA) and primarily repressing protein synthesis. Here, we describe miRNA and mRNA transcriptomic profiles in acute kidney injury (AKI). Further, we hypothesized that biobanked renal tissue biopsies (IB) with the pathologic diagnosis (path) of minimal change (MCD) or thin basement membrane (TBM) disease should demonstrate similar transcriptomic profiles to reference nephrectomies (Nx) and in comparisons with AKI.
Methods
AKI, IB, and nephrectomy (Nx) transcriptomic data (miR, RNA) were derived from stored human kidney tissues obtained in the routine clinical care of patients. AKI specimens carried path of acute interstitial or tubular necrosis (n= 39). IB specimens had path of MCD (n= 11) or TBM (n=1), IFTA <5% , no AKI. RNA was extracted from tissue (Qiagen miRNeasy), libraries prepared (RNA: Takara Total RNA-Seq, miR: Qiaseq) and sequenced. Differentially expressed genes (DEGs) and miRNAs (DEMs) were identified with DESeq2 using comparisons between: AKI/IB, AKI/Nx, IB/Nx. KEGG enrichment analysis was performed for DEGs and DEMs with FDR <0.05 using pathfindR for R, Mienturnet, and miRPathDBv2.0. MiRNA-mRNA interactions were identified using miRTarBase and miRDB.
Results
In AKI, increased miR-150-5p was associated with downregulation of IL1A, CTSB (related to MAPK signaling, metabolism). Similarly, increased miR-155-5p was associated with decreased HIF1a, known to exacerbate AKI. Next, we determined if IB can be used as a surrogate reference, and if each is distinct from AKI. In comparing the miRNA and mRNA transcriptomes of all samples, principal component analysis revealed 3 distinct clusters corresponding to IB, Nx and AKI. Comparing IB to Nx, enriched pathways included cell cycle, cancer pathways, and reactive oxygen species. AKI/Nx and AKI/IB comparisons enriched similarly in PI3K-Akt and related structural pathways (e.g. extracellular matrix, actin cytoskeleton, focal adhesion).
Conclusion
The miR and RNA targetome of AKI were similar when compared to both reference sets (IB and Nx). Our data suggest that miR regulate RNA in several important AKI-related pathways. Additionally, we propose the use of IB as tissue reference data for AKI.
Funding
- Other NIH Support