Elevated level of lysophosphatidic acid among patients with HNF1B mutations and its role in RCAD syndrome: a multiomic study

Joanna Madzio,Sławomir Chrul,Maciej Borowiec,Kamila Wypyszczak,Marcin Tkaczyk,Justyna Janikiewicz,Michał Ciborowski,Karolina Pietrowska,Adam Krętowski,Agnieszka Dobrzyń,Wojciech Fendler,Maciej T. Małecki,Anna Hogendorf,Wojciech Młynarski,Krystyna Wyka,Rafał Zwiech,Beata Małachowska

doi:10.1007/s11306-022-01873-z

Abstract

IntroductionPatients with hepatocyte nuclear factor-1 beta (HNF1B) mutations present a variable phenotype with two main symptoms: maturity onset diabetes of the young (MODY) and polycystic kidney disease (PKD).ObjectivesIdentification of serum metabolites specific for HNF1Bmut and evaluation of their role in disease pathogenesis.MethodsWe recruited patients with HNF1Bmut (N = 10), HNF1Amut (N = 10), PKD: non-dialyzed and dialyzed (N = 8 and N = 13); and healthy controls (N = 12). Serum fingerprinting was performed by LC-QTOF-MS. Selected metabolite was validated by ELISA (enzyme-linked immunosorbent assay) measurements and then biologically connected with HNF1B by in silico analysis. HepG2 were stimulated with lysophosphatidic acid (LPA) and HNF1B gene was knocked down (kd) by small interfering RNA. Transcriptomic analysis with microarrays and western blot measurements were performed.ResultsSerum levels of six metabolites including: arachidonic acid, hydroxyeicosatetraenoic acid, linoleamide and three LPA (18:1, 18:2 and 20:4), had AUC (the area under the curve) > 0.9 (HNF1Bmut vs comparative groups). The increased level of LPA was confirmed by ELISA measurements. In HepG2HNF1Bkd cells LPA stimulation lead to downregulation of many pathways associated with cell cycle, lipid metabolism, and upregulation of steroid hormone metabolism and Wnt signaling. Also, increased intracellular protein level of autotaxin was detected in the cells. GSK-3alpha/beta protein level and its phosphorylated ratio were differentially affected by LPA stimulation in HNF1Bkd and control cells.ConclusionsLPA is elevated in sera of patients with HNF1Bmut. LPA contributes to the pathogenesis of HNF1B-MODY by affecting Wnt/GSK-3 signaling.

Highlights

Patients with hepatocyte nuclear factor-1 beta (HNF1B) mutations present a variable phenotype with two main symptoms: maturity onset diabetes of the young (MODY) and polycystic kidney disease (PKD)
We showed that the obtained panel of 8 metabolites perfectly separated patients with HNF1Bmut from all other comparative groups (Fig. 1A)
In further studies, we focused on linking the lysophosphatidic acid (LPA) with HNF1B-MODY syndrome pathogenesis, searching for a source of higher LPA level in serum among HNF1Bmut patients and on studying the effect of higher LPA level on HNF1B deficient cells

Summary

Introduction

Patients with hepatocyte nuclear factor-1 beta (HNF1B) mutations present a variable phenotype with two main symptoms: maturity onset diabetes of the young (MODY) and polycystic kidney disease (PKD). Hepatocyte nuclear factor-1 beta (HNF1B)-MODY, called RCAD (renal cysts and diabetes syndrome) or MODY 5, is characterized by a variable coexistence of Extended author information available on the last page of the article diabetes and congenital abnormalities of kidneys and urinary tract (CAKUT). It is caused by an autosomal dominant mutation in HNF1B gene. It can bind to promoter DNA of target genes as a homodimer or heterodimer (with Hnf1a) leading to gene expression activation (mostly) or repression (rarely) (Musetti et al, 2014)

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