KCNJ16-depleted kidney organoids recapitulate tubulopathy and lipid recovery upon statins treatment

E Sendino Garví; G J J van Slobbe; E A Zaal; J H F de Baaij; J G Hoenderop; R Masereeuw; M J Janssen; A M van Genderen

doi:10.1186/s13287-024-03881-3

KCNJ16-depleted kidney organoids recapitulate tubulopathy and lipid recovery upon statins treatment

E Sendino Garví, G J J van Slobbe, E A Zaal, J H F de Baaij, J G Hoenderop, R Masereeuw, M J Janssen^*, A M van Genderen

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

BACKGROUND: The KCNJ16 gene has been associated with a novel kidney tubulopathy phenotype, viz. disturbed acid-base homeostasis, hypokalemia and altered renal salt transport. KCNJ16 encodes for Kir5.1, which together with Kir4.1 constitutes a potassium channel located at kidney tubular cell basolateral membranes. Preclinical studies provided mechanistic links between Kir5.1 and tubulopathy, however, the disease pathology remains poorly understood. Here, we aimed at generating and characterizing a novel advanced in vitro human kidney model that recapitulates the disease phenotype to investigate further the pathophysiological mechanisms underlying the tubulopathy and potential therapeutic interventions.

METHODS: We used CRISPR/Cas9 to generate KCNJ16 mutant (KCNJ16 +/- and KCNJ16 -/-) cell lines from healthy human induced pluripotent stem cells (iPSC) KCNJ16 control (KCNJ16 WT). The iPSCs were differentiated following an optimized protocol into kidney organoids in an air-liquid interface.

RESULTS: KCNJ16-depleted kidney organoids showed transcriptomic and potential functional impairment of key voltage-dependent electrolyte and water-balance transporters. We observed cysts formation, lipid droplet accumulation and fibrosis upon Kir5.1 function loss. Furthermore, a large scale, glutamine tracer flux metabolomics analysis demonstrated that KCNJ16 -/- organoids display TCA cycle and lipid metabolism impairments. Drug screening revealed that treatment with statins, particularly the combination of simvastatin and C75, prevented lipid droplet accumulation and collagen-I deposition in KCNJ16 -/- kidney organoids.

CONCLUSIONS: Mature kidney organoids represent a relevant in vitro model for investigating the function of Kir5.1. We discovered novel molecular targets for this genetic tubulopathy and identified statins as a potential therapeutic strategy for KCNJ16 defects in the kidney.

Original language	English
Article number	268
Journal	Stem cell research & therapy
Volume	15
Issue number	1
DOIs	https://doi.org/10.1186/s13287-024-03881-3
Publication status	Published - 26 Aug 2024

Bibliographical note

Publisher Copyright:
© The Author(s) 2024.

Funding

This research received funding from the IMAGEN project which is co-funded by the PPP Allowance made available by Health\u2009~\u2009Holland, Top Sector Life Sciences & Health, to stimulate public\u2013private partnerships (IMplementation of Advancements in GENetic Kidney Disease, LSHM20009; ESG, MJJ, and RM) and from the Netherlands Research Council (NWO) \u2018Netherlands Research Agenda: Research on Routes by Consortia\u2019 (NWA-ORC 1292.19.272) Virtual Human Platform for Safety consortium (AMG, RM). The authors would like to thank Sam Biermans for the cell culture support and the valuable discussions about the results of this manuscript. The authors wish to thank the Radboudumc Stem Cell Technology and differentiation Center (https://www.radboudumc.nl/en/research/radboud-technology-centers/stem-cells) for reprogramming and characterizing the cell lines.

Funders	Funder number
Nederlandse Organisatie voor Wetenschappelijk Onderzoek
Radboudumc Stem Cell Technology and differentiation Center
Netherlands Research Council
Health ~ Holland, Top Sector Life Sciences & Health	LSHM20009
NWA-ORC	1292.19.272

Keywords

Humans
Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology
Induced Pluripotent Stem Cells/metabolism
Kidney/metabolism
Lipid Metabolism/drug effects
Organoids/metabolism
Potassium Channels, Inwardly Rectifying/metabolism

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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s13287-024-03881-3Final published version, 17.2 MBLicence: CC BY

Cite this

@article{6d35287cdc774693aa258a6ff6684457,

title = "KCNJ16-depleted kidney organoids recapitulate tubulopathy and lipid recovery upon statins treatment",

abstract = "BACKGROUND: The KCNJ16 gene has been associated with a novel kidney tubulopathy phenotype, viz. disturbed acid-base homeostasis, hypokalemia and altered renal salt transport. KCNJ16 encodes for Kir5.1, which together with Kir4.1 constitutes a potassium channel located at kidney tubular cell basolateral membranes. Preclinical studies provided mechanistic links between Kir5.1 and tubulopathy, however, the disease pathology remains poorly understood. Here, we aimed at generating and characterizing a novel advanced in vitro human kidney model that recapitulates the disease phenotype to investigate further the pathophysiological mechanisms underlying the tubulopathy and potential therapeutic interventions.METHODS: We used CRISPR/Cas9 to generate KCNJ16 mutant (KCNJ16 +/- and KCNJ16 -/-) cell lines from healthy human induced pluripotent stem cells (iPSC) KCNJ16 control (KCNJ16 WT). The iPSCs were differentiated following an optimized protocol into kidney organoids in an air-liquid interface. RESULTS: KCNJ16-depleted kidney organoids showed transcriptomic and potential functional impairment of key voltage-dependent electrolyte and water-balance transporters. We observed cysts formation, lipid droplet accumulation and fibrosis upon Kir5.1 function loss. Furthermore, a large scale, glutamine tracer flux metabolomics analysis demonstrated that KCNJ16 -/- organoids display TCA cycle and lipid metabolism impairments. Drug screening revealed that treatment with statins, particularly the combination of simvastatin and C75, prevented lipid droplet accumulation and collagen-I deposition in KCNJ16 -/- kidney organoids. CONCLUSIONS: Mature kidney organoids represent a relevant in vitro model for investigating the function of Kir5.1. We discovered novel molecular targets for this genetic tubulopathy and identified statins as a potential therapeutic strategy for KCNJ16 defects in the kidney.",

keywords = "Humans, Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology, Induced Pluripotent Stem Cells/metabolism, Kidney/metabolism, Lipid Metabolism/drug effects, Organoids/metabolism, Potassium Channels, Inwardly Rectifying/metabolism",

author = "{Sendino Garv{\'i}}, E and {van Slobbe}, {G J J} and Zaal, {E A} and {de Baaij}, {J H F} and Hoenderop, {J G} and R Masereeuw and Janssen, {M J} and {van Genderen}, {A M}",

note = "Publisher Copyright: {\textcopyright} The Author(s) 2024.",

year = "2024",

month = aug,

day = "26",

doi = "10.1186/s13287-024-03881-3",

language = "English",

volume = "15",

journal = "Stem cell research & therapy",

issn = "1757-6512",

publisher = "BioMed Central",

number = "1",

}

TY - JOUR

T1 - KCNJ16-depleted kidney organoids recapitulate tubulopathy and lipid recovery upon statins treatment

AU - Sendino Garví, E

AU - van Slobbe, G J J

AU - Zaal, E A

AU - de Baaij, J H F

AU - Hoenderop, J G

AU - Masereeuw, R

AU - Janssen, M J

AU - van Genderen, A M

PY - 2024/8/26

Y1 - 2024/8/26

N2 - BACKGROUND: The KCNJ16 gene has been associated with a novel kidney tubulopathy phenotype, viz. disturbed acid-base homeostasis, hypokalemia and altered renal salt transport. KCNJ16 encodes for Kir5.1, which together with Kir4.1 constitutes a potassium channel located at kidney tubular cell basolateral membranes. Preclinical studies provided mechanistic links between Kir5.1 and tubulopathy, however, the disease pathology remains poorly understood. Here, we aimed at generating and characterizing a novel advanced in vitro human kidney model that recapitulates the disease phenotype to investigate further the pathophysiological mechanisms underlying the tubulopathy and potential therapeutic interventions.METHODS: We used CRISPR/Cas9 to generate KCNJ16 mutant (KCNJ16 +/- and KCNJ16 -/-) cell lines from healthy human induced pluripotent stem cells (iPSC) KCNJ16 control (KCNJ16 WT). The iPSCs were differentiated following an optimized protocol into kidney organoids in an air-liquid interface. RESULTS: KCNJ16-depleted kidney organoids showed transcriptomic and potential functional impairment of key voltage-dependent electrolyte and water-balance transporters. We observed cysts formation, lipid droplet accumulation and fibrosis upon Kir5.1 function loss. Furthermore, a large scale, glutamine tracer flux metabolomics analysis demonstrated that KCNJ16 -/- organoids display TCA cycle and lipid metabolism impairments. Drug screening revealed that treatment with statins, particularly the combination of simvastatin and C75, prevented lipid droplet accumulation and collagen-I deposition in KCNJ16 -/- kidney organoids. CONCLUSIONS: Mature kidney organoids represent a relevant in vitro model for investigating the function of Kir5.1. We discovered novel molecular targets for this genetic tubulopathy and identified statins as a potential therapeutic strategy for KCNJ16 defects in the kidney.

AB - BACKGROUND: The KCNJ16 gene has been associated with a novel kidney tubulopathy phenotype, viz. disturbed acid-base homeostasis, hypokalemia and altered renal salt transport. KCNJ16 encodes for Kir5.1, which together with Kir4.1 constitutes a potassium channel located at kidney tubular cell basolateral membranes. Preclinical studies provided mechanistic links between Kir5.1 and tubulopathy, however, the disease pathology remains poorly understood. Here, we aimed at generating and characterizing a novel advanced in vitro human kidney model that recapitulates the disease phenotype to investigate further the pathophysiological mechanisms underlying the tubulopathy and potential therapeutic interventions.METHODS: We used CRISPR/Cas9 to generate KCNJ16 mutant (KCNJ16 +/- and KCNJ16 -/-) cell lines from healthy human induced pluripotent stem cells (iPSC) KCNJ16 control (KCNJ16 WT). The iPSCs were differentiated following an optimized protocol into kidney organoids in an air-liquid interface. RESULTS: KCNJ16-depleted kidney organoids showed transcriptomic and potential functional impairment of key voltage-dependent electrolyte and water-balance transporters. We observed cysts formation, lipid droplet accumulation and fibrosis upon Kir5.1 function loss. Furthermore, a large scale, glutamine tracer flux metabolomics analysis demonstrated that KCNJ16 -/- organoids display TCA cycle and lipid metabolism impairments. Drug screening revealed that treatment with statins, particularly the combination of simvastatin and C75, prevented lipid droplet accumulation and collagen-I deposition in KCNJ16 -/- kidney organoids. CONCLUSIONS: Mature kidney organoids represent a relevant in vitro model for investigating the function of Kir5.1. We discovered novel molecular targets for this genetic tubulopathy and identified statins as a potential therapeutic strategy for KCNJ16 defects in the kidney.

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KW - Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology

KW - Induced Pluripotent Stem Cells/metabolism

KW - Kidney/metabolism

KW - Lipid Metabolism/drug effects

KW - Organoids/metabolism

KW - Potassium Channels, Inwardly Rectifying/metabolism

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U2 - 10.1186/s13287-024-03881-3

DO - 10.1186/s13287-024-03881-3

M3 - Article

C2 - 39183338

SN - 1757-6512

VL - 15

JO - Stem cell research & therapy

JF - Stem cell research & therapy

IS - 1

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ER -

KCNJ16-depleted kidney organoids recapitulate tubulopathy and lipid recovery upon statins treatment

Abstract

Bibliographical note

Funding

Keywords

UN SDGs

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