A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Human pluripotent stem cell-derived cells endogenously expressing follicle-stimulating hormone receptors: modeling the function of an inactivating receptor mutation
Tekijät: Lundin K, Sepponen K, Väyrynen P, Liu X, Yohannes DA, 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 JS
Kustantaja: OXFORD UNIV PRESS
Julkaisuvuosi: 2022
Journal: Molecular Human Reproduction
Tietokannassa oleva lehden nimi: MOLECULAR HUMAN REPRODUCTION
Lehden akronyymi: MOL HUM REPROD
Artikkelin numero: gaac012
Vuosikerta: 28
Numero: 5
Sivujen määrä: 13
ISSN: 1360-9947
eISSN: 1460-2407
DOI: https://doi.org/10.1093/molehr/gaac012
Verkko-osoite: https://academic.oup.com/molehr/article/28/5/gaac012/6574364
Rinnakkaistallenteen osoite: https://research.utu.fi/converis/portal/detail/Publication/175412513
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.
Ladattava julkaisu This is an electronic reprint of the original article. |  |
