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Tacrolimus-induced nephrotoxicity in tubuloids

Tacrolimus (TAC) is a first-line immunosuppressant used to prevent rejection after solid-organ transplantation, yet its clinical utility is limited by nephrotoxicity, which can progress to irreversible kidney injury or graft loss. The molecular mechanisms underlying TAC-induced nephrotoxicity remain insufficiently understood. Here, we systematically assessed TAC responses in a human in vitro nephron model: adult stem cell-derived kidney tubuloids from two donors (T19 and T30). Cell viability (CellTiter-Fluor), intracellular tacrolimus accumulation (LC-MS/MS), targeted gene expression of drug transporters and metabolizing enzymes (RT-qPCR) and whole-transcriptome responses (bulk RNA-seq) were measured, with co-treatment by P-glycoprotein (P-gp) inhibitors used to modulate intracellular tacrolimus levels.

Details
Type
study
Internal ID
S-VHPS25
Release date
2026-03-18
Version
—
License
CC-BY
URL
https://www.ebi.ac.uk/biostudies/studies/S-VHPS25
DOI
10.6019/S-VHPS25
Publications

No linked publications available.

Files

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Authors
Funding
  • Dutch Research Agenda (NWA)
    1292.19.272
Human kidney tubuloids
RRID: N/A · Supplier: Established in-house from human adult kidney tissue.
Homo sapiens · Kidney · Nephron tubule epithelium · Kidney tubular epithelial cell

Adult stem cell-derived kidney tubuloids established from human kidney tissue of two donors (T19 and T30); expanded and differentiated in vitro to model the renal tubular epithelium. Donor demographics (age, sex, diagnosis, tissue origin) are not documented in the source data.

Exposure concentration: 16-104 (12-point linear series, 8 µM steps); LC-MS quantified an 8-point subset µM · Co-treatment: ± P-gp inhibitor (PSC-833, 5 µM)
Exposure-measurement interval: 0 hours · Vehicle: DMSO · Exposure duration: 24 hours
Bioassays: CellTiter-Fluor, LCMS
Exposure concentration: 16, 40 (RNA-seq also tested 64, excluded as outlier) µM
Exposure-measurement interval: 0 hours · Vehicle: DMSO · Exposure duration: 24 hours
Bioassays: RT-qPCR, RNA-sequencing
Bioassays
  • CellTiter-Fluor [DRAFT]
    Measured: Fluorescence · Readout: Cell viability (% of vehicle control) · Replicates: ≥3 · Positive control: Hydrogen peroxide
    Vehicle control: DMSO
  • LCMS [DRAFT]
    Measured: Tacrolimus (analyte/IS peak-area ratio) · Readout: Intracellular tacrolimus (ng/mg protein) · Replicates: ≥3 · Technical replicates: 3
    Internal standard: Sirolimus
  • RT-qPCR MESH
    Measured: mRNA · Readout: Relative gene expression (fold-change vs control) · Replicates: [DRAFT]
    Reference gene: β-actin (ACTB)
  • RNA-sequencing [DRAFT]
    Measured: Transcript abundance · Readout: Differentially expressed genes (DEGs) (log2 fold-change) · Replicates: 2
AOP Linkage

Coming soon: normalization, model fitting, statistics, QC, etc.

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Kidney Question 2: Tacrolimus toxicity Toxicodynamics