Kidney Week

Abstract: TH-PO0571

PTH1R Signaling Instigates a Cystogenic Program in Autosomal Recessive PKD Organoids

Session Information

• Development, Stem Cells, and Regenerative Medicine
  November 06, 2025 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Genetic Diseases of the Kidneys

• 1201 Genetic Diseases of the Kidneys: Monogenic Kidney Diseases

Authors

• Afrin, Humayra, Mayo Clinic Minnesota, Rochester, Minnesota, United States

Humayra Afrin, MD, PhD

• Robichaud, Jielu Hao, Mayo Clinic Minnesota, Rochester, Minnesota, United States

Jielu Hao Robichaud, MD, PhD

• Qamar, Usama, Mayo Clinic Minnesota, Rochester, Minnesota, United States

Usama Qamar, MD

• Harris, Peter C., Mayo Clinic Minnesota, Rochester, Minnesota, United States

Peter C. Harris, PhD

• Gupta, Navin R., Mayo Clinic Minnesota, Rochester, Minnesota, United States

Navin R. Gupta, MD

Background

Autosomal recessive polycystic kidney disease (ARPKD) is an inherited developmental cystic kidney disease caused by biallelic mutations in PKHD1. ARPKD confers high morbidity and mortality due to a lack of directed therapies. Unfortunately, the various Pkhd1 mutant mouse models (Pkhd1^LSL/LSL, Pkhd1^Δ3-4/Δ3-4, and Pkhd1^Δ67/Δ67) fail to recapitulate the human developmental phenotype, manifesting limited renal disease. To overcome this limitation and to stimulate drug discovery, human pluripotent stem cell (hPSC)-derived ARPKD organoids have manifested well-described cystogenic mechanisms suggesting value for therapeutic screens. In these studies, PKHD1^-/- organoids have been challenged with forskolin to induce cyclic AMP, a hallmark of PKD. While these studies validate genotype-phenotype correlation and the faithfulness of cystic responses, novel cystogenic mechanisms that expand potential therapeutic targets are yet to be reported.

Methods

We use parental wild-type human PGP1 and isogenic CRISPR PKHD1^-/- iPSCs, directed differentiation to kidney organoids, longitudinal brightfield microscopy, wholemount and cryosection confocal microscopy, native kidney and organoid single cell RNAseq, ARPKD tissue, and targeted mechanistic studies to describe PTH1R as a stimulatory G-protein coupled receptor (sGPCR) which instigates a cystogenic signaling cascade in developmental cystic kidney disease.

Results

PTHR1 loss was confirmed in ARPKD organoids by immunostaining. Chemical cAMP induction in ARPKD organoids induced a dose-dependent cystic response, unlike their isogenic wild-type control, prompting a search for upstream sGPCRs as a driver for cAMP. Unbiased transcriptomic screen of native human kidney for sGPCRs identified PTH1R, which co-localized with LTL⁺ and CDH1⁺ in proximal and distal tubule cysts of ARPKD patient kidney tissue. ARPKD organoids subject to endogenous ligands of differential affinity for parathyroid hormone receptors recognized PTH1R agonism as a novel cystogenic stimuli, with cysts having apical-facing primary cilia. Targeted inhibition of adenylyl cyclase and Protein Kinase A demonstrated that the stimulatory GPCR, PTH1R, drives a cystogenic cAMP-PKA-CREB axis in developmental PKD.

Conclusion

Our findings demonstrate the utility of kidney organoids as an in vitro model to explore pathomechanisms of rare diseases which lack faithful animal models.

Funding

• NIDDK Support

Digital Object Identifier (DOI)

• doi: 10.1681/ASN.2025nypk18g3