Integrative transcriptomic profiling of a mouse model of hypertension-accelerated diabetic kidney disease

Abstract

The current understanding of molecular mechanisms driving diabetic kidney disease (DKD) is limited, partly due to the complex structure of the kidney. To identify genes and signalling pathways involved in the progression of DKD, we compared kidney cortical vs. glomerular transcriptome profiles in uninephrectomized (UNx) db/db mouse models of early-stage (UNx only) and advanced (UNx plus AAV-mediated renin overexpression, UNx-Renin) DKD using RNA sequencing (RNAseq). Compared to normoglycemic db/m mice, db/db UNx and db/db UNx-Renin mice showed marked changes in kidney cortical and glomerular gene expression profiles. UNx-Renin mice displayed more marked perturbations in gene components associated with activation of the immune system and enhanced extracellular matrix remodelling, supporting histological hallmarks of progressive DKD in this model. Single-nucleus RNAseq enabled linking transcriptome profiles to specific kidney cell types. In conclusion, integration of RNAseq at the cortical, glomerular and single-nucleus level provides enhanced resolution of molecular signalling pathways associated with disease progression in preclinical models of DKD, and may thus be advantageous for identifying novel therapeutic targets in DKD.

Nephroprotective Effects of Semaglutide as Mono- and Combination Treatment with Lisinopril in a Mouse Model of Hypertension-Accelerated Diabetic Kidney Disease

“…This open access resource can be used for further data mining on global gene expression changes. Cell type deconvolution was based on single-nucleus RNA sequencing data previously reported in the db / db UNx-ReninAAV mouse [ 15 ]. Individual DEGs obtained from bulk RNA sequencing analysis were assigned to kidney cell types predominantly expressing the corresponding regulated gene ( Figure 7 B, individual DEGs are listed in Table S1 ).…”

“…Gene set analysis was conducted with the R package PIANO version 1.18.1 using the Stouffer method, and p -values were corrected for multiple testing using the Benjamini–Hochberg method (5% false discovery rate, FDR < 0.05). To obtain further resolution of gene expression regulations, expression of treatment-regulated genes was evaluated across kidney cell types using single-nuclei sequencing data from db / db UNx-ReninAAV mice, with data processed as described previously [ 15 ]. For clustering analysis, the geometric mean of relative counts +1 was calculated for each cell type, and mean values were centred and scaled prior to hierarchical clustering.…”

“…A state-of-the-art translational mouse model of hypertension-accelerated DKD has recently been established, induced by adeno-associated virus (AAV)-mediated renin overexpression in the uninephrectomized (UNx) diabetic db / db ( db / db UNx-ReninAAV) mouse [ 13 , 14 , 15 ]. db / db UNx-ReninAAV mice display key features of advanced diabetic kidney disease, including extreme albuminuria and severe glomerulosclerosis, which can be ameliorated by current clinical standard of care using ACE inhibitor (lisinopril) and SGLT2i (empagliflozin) combination therapy [ 13 ].…”

What lies beyond 100 years of insulin

“…As there is currently no cure for diabetes, this life-long condition manifests itself in complications affecting organs beyond the pancreas and liver, such as the kidneys, and the peripheral and central nervous systems. Diabetic kidney disease (DKD) accounts for 25–50% of patients with kidney failure in Europe and the USA ( Sembach et al, 2021 ). Advances in transcriptomic profiling have enabled deeper investigation of the molecular signalling pathways involved in development and progression of DKD in rodent models ( Otto et al, 2019 ; Sembach et al, 2021 ), which is essential to identify novel targets for preventing or halting DKD.…”

“…Diabetic kidney disease (DKD) accounts for 25–50% of patients with kidney failure in Europe and the USA ( Sembach et al, 2021 ). Advances in transcriptomic profiling have enabled deeper investigation of the molecular signalling pathways involved in development and progression of DKD in rodent models ( Otto et al, 2019 ; Sembach et al, 2021 ), which is essential to identify novel targets for preventing or halting DKD. Peripheral neuropathy (PN) is a complication that occurs in 30% of prediabetes and 60% of T2D patients, and can lead to chronic pain and lower-limb amputations.…”