Lundin, K., Sepponen, K., Väyrynen, P., Liu, X., Yohannes, D. A., Survila, M., Ghimire, B., Känsäkoski, J., Katayama, S., Partanen, J., Vuoristo, S., Paloviita, P., Rahman, N., Raivio, T., Luiro, K., Huhtaniemi, I., Varjosalo, M., Tuuri, T., & Tapanainen, J. S. (2022). Human pluripotent stem cell-derived cells endogenously expressing follicle-stimulating hormone receptors: Modeling the function of an inactivating receptor mutation. Molecular Human Reproduction, 28(5), gaac012. https://doi.org/10.1093/molehr/gaac012
Human pluripotent stem cell-derived cells endogenously expressing follicle-stimulating hormone receptors : modeling the function of an inactivating receptor mutation
|Author:||Lundin, K.1; Sepponen, K.1; Väyrynen, P.1;|
1Department of Obstetrics and Gynecology, University of Helsinki and Helsinki University Hospital, Helsinki, Finland
2Molecular Systems Biology Research Group, Institute of Biotechnology & HiLIFE, University of Helsinki, Helsinki, Finland
3Proteomics Unit, Institute of Biotechnology & HiLIFE, University of Helsinki, Helsinki, Finland
4Research Programs Unit, Translational Immunology & Department of Medical and Clinical Genetics, University of Helsinki, Helsinki, Finland
5Molecular and Integrative Biosciences Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, Helsinki, Finland
6Institute for Molecular Medicine Finland, University of Helsinki, Helsinki, Finland
7Department of Physiology, University of Helsinki, Helsinki, Finland
8Folkhälsan Research Center, Helsinki, Finland
9Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Sweden
10Stem Cells and Metabolism Research Program, Faculty of Medicine, University of Helsinki, Helsinki, Finland
11Institute of Biomedicine, University of Turku, Turku, Finland
12Department of Reproduction and Gynecological Endocrinology, Medical University of Bialystok, Bialystok, Poland
13New Children's Hospital, Pediatric Research Center, Helsinki University Hospital, HUH, Helsinki, Finland
14Department of Metabolism, Endocrinology and Reproduction, Faculty of Medicine, Hammersmith Campus, Imperial College London, London, UK
15Department of Obstetrics and Gynecology, University Hospital of Oulu, University of Oulu, Medical Research Center Oulu and PEDEGO Research Unit, Oulu, Finland
|Online Access:||PDF Full Text (PDF, 1.5 MB)|
|Persistent link:|| http://urn.fi/urn:nbn:fi-fe2022112466835
Oxford University Press,
|Publish Date:|| 2022-11-24
Follicle-stimulating hormone (FSH) is crucial in the development and regulation of reproductive functions. The actions of human FSH and its receptor (FSHR) and mutations therein have mainly been studied using in vivo models, primary cells, cancer cells and cell lines ectopically expressing the FSHR. To allow studies of endogenous FSHR function in vitro, we differentiated FSHR-expressing cells from human pluripotent stem cells. FSH stimulation of the wild-type (WT), but not the inactivating Finnish founder mutant (A189V) receptor, activated the canonical cyclic adenosine monophosphate (cAMP)-dependent signaling pathway and downstream mediators. To investigate protein–protein interaction partners of FSHR at resting state and upon FSH stimulation, we expressed FSHR in HEK293 cells followed by affinity purification mass spectrometry analyses. We found 19 specific high-confidence interacting proteins for WT FSHR and 14 for A189V FSHR, several of which have been linked to infertility. Interestingly, while only WT FSHR interacted with FSH, insulin-like growth factor 1 receptor (IGF1R), for example, interacted with both WT and A189V FSHR upon FSH stimulation. In conclusion, our protocol allows detailed studies of FSH action and disease modeling in human cells endogenously expressing FSHR.
Molecular human reproduction
|Type of Publication:||
A1 Journal article – refereed
|Field of Science:||
3123 Gynaecology and paediatrics
This study was financially supported by the Academy of Finland (Grant 295760; J.S.T.), Novo Nordisk Fonden (Grant 10825; J.S.T.), Helsinki University Hospital Funds (J.S.T. and T.T.), the Sigrid Juselius Foundation (J.S.T.), the Finnish Medical Foundation (K.Lui.) and Jane and Aatos Erkko Foundation (S.K.).
Single-cell RNA sequencing data and expression matrix of filtered and normalized single-cell RNA sequencing data has been deposited in NCBI’s Gene Expression Omnibus (GEO) and can be accessed through GEO Series accession number GSE184352 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE184352). Mass spectrometry data has been deposited to MassIVE (https://massive.ucsd.edu/) with the identifier: MSV000088577.
© The Author(s) 2022. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.