Modeling HNF1B-associated monogenic diabetes using human iPSCs reveals an early stage impairment of the pancreatic developmental program

OLSZANOWSKI, E.; EL-KHAIRI, R.; MURARO, D.; MADRIGAL, P.; LUDOVIC VALLIER; RODRÍGUEZ SEGUÍ, S.A.

Congreso Virtual

Congreso; XI Congreso Argentino de Bioinformática y Biología Computacional; 2021

Asociación Argentina de Bioinformática y Biología Computacional (A2B2C)

Background:Maturity-onsetdiabetes of the young (MODY) is the most common form of monogenicdiabetes, and it is characterized by autosomal dominant inheritance,and hyperglycemia due to β cell failure. Hepatic nuclear factor 1b(HNF1B), plays an important role in the normal development of thekidney, liver, pancreas, bile ducts, and urogenital tract, throughtissue-specific regulation of gene expression in these organs. Inhumans, heterozygous mutations in HNF1B result in a multisystemdisorder, associated with MODY5. Results:BulkRNA-seq experiments of HNF1B-/-(1βHom)hiPSC-derived progenitors at the FP stage (day 6), and at posteriorforegut (day 8) revealed upregulationinalternative developmental pathways, notably heart, kidney, andnervous system development, showing the central role of HNF1B in thespecification of the foregut toward the pancreatic lineages. In all1βHetand 1βHomsamples from all stages we found a consistent downregulation of theHNF1A antisense long non-coding RNA (lncRNA HNF1A-AS1). Single-cellRNA-seq on HNF1B+/+(1βWT)and HNF1B+/-(1βHet)cells from day 13 revealed that the number of less proliferative(late) MPCs in D13-1βHetsamples was higher than in their 1βWTcounterparts, and this increase appeared to take place mainly at theexpense of the highly proliferative (early) MPC population, whichexpress increased levels of the proliferation markers TOP2A andAURKB. These results suggest that HNF1B plays an important role inallowing the proliferative early MPC stage.Conclusions:Ourresults show that lack of HNF1B blocks specification of pancreaticfate from the foregut progenitor (FP) stage, but HNF1Bhaploinsufficiency allows differentiation of multipotent pancreaticprogenitor cells (MPCs) and insulin-secreting b-like cells. Weshow that HNF1B plays a central role in the specification of theforegut towards the pancreatic lineages by controlling key masterregulators; the absence of HNF1B affects foregut patterning byallowing cells to adopt alternative fates. Also, we found that HNF1Bhaploinsufficiency impairs cell proliferation in FPs and MPCs. Thiscould be attributed to impaired induction of key pancreaticdevelopmental genes, including SOX11, ROBO2, and additional TEAD1target genes whose function is associated with MPC self-renewal. p { margin-bottom: 0.1in; direction: ltr; color: #000000; line-height: 115%; text-align: left; orphans: 2; widows: 2 }p.western { font-family: "Liberation Serif", serif; font-size: 12pt; so-language: es-AR }p.cjk { font-family: "Noto Serif CJK SC"; font-size: 12pt; so-language: zh-CN }p.ctl { font-family: "Lohit Devanagari"; font-size: 12pt; so-language: hi-IN }