FC 001ACIDOSIS AND ALKALI THERAPY ARE ASSOCIATED WITH TRANSCRIPTIONAL CHANGES AND ALTERED ABUNDANCE OF GENES INVOLVED IN CELL METABOLISM AND BICARBONATE TRANSPORT IN KIDNEY TRANSPLANT RECIPIENTS
Authors: Imenez Silva Pedro Henrique , Wiegand Anna , Daryadel Arezoo , Gaspert Ariana , Russo Giancarlo , Wuthrich Rudolf Peter , Wagner Carsten , Mohebbi Nilufar 
Published: May 2021

Abstract

Background and Aims
Metabolic acidosis is a common event in kidney transplant recipients and has been associated to a higher risk of graft loss and mortality. In patients with CKD and acidosis, alkali therapy ameliorating acidosis appears to protect kidney function. However, it is still poorly understood how acidosis causes the detrimental effects to kidney graft function and how alkali therapy would interact with these mechanisms. Here we aim to identify transcriptomic alterations in kidney transplant recipients without metabolic acidosis in comparison to patients with metabolic acidosis with and without alkali therapy. Moreover, we examined immunolocalization of key proteins involved in acid-base base regulation in biopsies from these patients.

Method
We obtained 22 biopsies of patients 4-6 years after kidney transplantation. Among these patients, nine were not acidotic (serum [HCO3-] ≥ 22 mM), nine had acidosis ([HCO3-] &lt; 22 mM), and four had acidosis and received sodium bicarbonate (alkali therapy) fully correcting acidosis. Age, immunosuppressive drugs, time after transplantation, and eGFR were not statistically different between groups. RNA was extracted from biopsies and RNAseq was performed. Immunohistochemistry was performed for key proteins involved in the renal regulation of acid-base balance. Additionally, a control group of 6 non-transplanted healthy kidneys was included in the histology analysis.

Results
RNAseq analysis revealed 40 genes differentially expressed between acidosis and no acidosis groups. While most of the genes tended to be recovered by alkali therapy, only three fully recovered with bicarbonate supplementation (p-value &lt; 0.05 and log2(fold change) above 0.5). These genes were KCNJ15 (Kir4.2), SHMT1, and ACADSB. Renal localization of the genes was determined using single-cell RNA sequencing data (Ransick et al., Developmental Cell, 2019, doi.org/10.1016/j.devcel.2019.10.005). Most of the genes were expressed in the proximal tubule and were organized in the model shown in Figure 1A. Several of these genes participate in cell metabolism, such as beta-oxidation, and iron, folate, and methionine metabolism. Moreover, the K+-channel Kir4.2 regulates the activity of the electrogenic sodium bicarbonate cotransporter 1 (NBCe1, SLC4A4) and ammoniagenesis in renal proximal tubules. Immunofluorescence analysis showed that NBCe1 expression in proximal tubules was strongly reduced in patients who developed acidosis and was partially recovered in patients who received alkali therapy (Figure 1B). In type B intercalated cells, a similar pattern was observed for Pendrin (SLC26A4). No alteration in the expression of GDH (GLUD1), AE1 (SLC4A1), AQP2, CA2, RhCG (SLC42A3), and B1 subunit of the H+ATPase (ATP6V1B1) was observed in kidneys of treated or untreated patients with acidosis.

Conclusion
Kidney transplant recipients suffering from metabolic acidosis show distinct expression pattern of genes involved in cell metabolism and acid-base transport.