DataSheet_1_Follicle-Stimulating Hormone Induces Lipid Droplets via Gαi/o and β-Arrestin in an Endometrial Cancer Cell Line.pdf
Follicle-stimulating hormone (FSH) and its G protein-coupled receptor, FSHR, represents a paradigm for receptor signaling systems that activate multiple and complex pathways. Classically, FSHR activates Gαs to increase intracellular levels of cAMP, but its ability to activate other G proteins, and β-arrestin-mediated signaling is well documented in many different cell systems. The pleiotropic signal capacity of FSHR offers a mechanism for how FSH drives multiple and dynamic downstream functions in both gonadal and non-gonadal cell types, including distinct diseases, and how signal bias may be achieved at a pharmacological and cell system-specific manner. In this study, we identify an additional mechanism of FSH-mediated signaling and downstream function in the endometrial adenocarcinoma Ishikawa cell line. While FSH did not induce increases in cAMP levels, this hormone potently activated pertussis toxin sensitive Gαi/o signaling. A selective allosteric FSHR ligand, B3, also activated Gαi/o signaling in these cells, supporting a role for receptor-mediated activation despite the low levels of FSHR mRNA. The low expression levels may attribute to the lack of Gαs/cAMP signaling as increasing FSHR expression resulted in FSH-mediated activation of the Gαs pathway. Unlike prior reports for FSH-mediated Gαs/cAMP signaling, FSH-mediated Gαi/o signaling was not affected by inhibition of dynamin-dependent receptor internalization. While chronic FSH did not alter cell viability, FSH was able to increase lipid droplet size. The β-arrestins are key adaptor proteins known to regulate FSHR signaling. Indeed, a rapid, FSH-dependent increase in interactions between β-arrestin1 and Gαi1 was observed via NanoBiT complementation in Ishikawa cells. Furthermore, both inhibition of Gαi/o signaling and siRNA knockdown of β-arrestin 1/2 significantly reduced FSH-induced lipid droplet accumulation, implying a role for a Gαi/o/β-arrestin complex in FSH functions in this cell type. As FSH/FSHR has been implicated in distinct hormone-dependent cancers, including endometrial cancer, analysis of the cancer genome database from 575 human endometrial adenocarcinoma tumors revealed that a subpopulation of samples expressed FSHR. Overall, this study highlights a novel mechanism for FSHR signal pleiotropy that may be exploited for future personalized therapeutic approaches.
History
References
- https://doi.org//10.1016/j.metabol.2017.11.018
- https://doi.org//10.1124/mol.117.111062
- https://doi.org//10.1111/j.1476-5381.2011.01434.x
- https://doi.org//10.1038/nrc3521
- https://doi.org//10.1371/journal.pbio.3000434
- https://doi.org//10.1074/jbc.AC120.014698
- https://doi.org//10.1016/j.jmb.2010.12.034
- https://doi.org//10.1038/ncomms9202
- https://doi.org//10.1073/pnas.1701529114
- https://doi.org//10.1038/s41594-018-0071-3
- https://doi.org//10.1016/j.cell.2020.02.047
- https://doi.org//10.1074/jbc.M806124200
- https://doi.org//10.1371/journal.pone.0028723
- https://doi.org//10.1073/pnas.1205756110
- https://doi.org//10.1074/jbc.RA120.015400
- https://doi.org//10.1073/pnas.2026491118
- https://doi.org//10.1126/science.aay1833
- https://doi.org//10.1124/pr.113.008052
- https://doi.org//10.1016/bs.ircmb.2018.03.001
- https://doi.org//10.1073/pnas.1905993116
- https://doi.org//10.1021/bi034907w
- https://doi.org//10.1210/me.2006-0098
- https://doi.org//10.1016/j.mce.2010.08.016
- https://doi.org//10.1096/fj.201700763R
- https://doi.org//10.1210/en.2013-1407
- https://doi.org//10.1038/s41598-018-20722-5
- https://doi.org//10.1016/j.isci.2020.101812
- https://doi.org//10.1074/jbc.M113.526350
- https://doi.org//10.3389/fphar.2020.593492
- https://doi.org//10.1016/j.cell.2006.01.051
- https://doi.org//10.3389/fendo.2011.00045
- https://doi.org//10.1111/acel.12331
- https://doi.org//10.1210/jendso/bvaa019
- https://doi.org//10.1093/humrep/deab135
- https://doi.org//10.1210/en.2018-00452
- https://doi.org//10.3389/fendo.2019.00305
- https://doi.org//10.3389/fendo.2019.00136
- https://doi.org//10.1210/jc.2013-3186
- https://doi.org//10.1095/biolreprod.114.118562
- https://doi.org//10.1371/journal.pone.0134986
- https://doi.org//10.1095/biolreprod.116.141648
- https://doi.org//10.1038/nature22342
- https://doi.org//10.1007/s10815-018-1248-8
- https://doi.org//10.3389/fendo.2019.00032
- https://doi.org//10.1016/S0022-5347%2801%2961831-7
- https://doi.org//10.1074/jbc.M003206200
- https://doi.org//10.1054/bjoc.2000.1507
- https://doi.org//10.1016/j.mce.2006.11.010
- https://doi.org//10.1016/j.bbacli.2016.11.002
- https://doi.org//10.1210/jc.2016-1014
- https://doi.org//10.1016/j.mce.2016.08.009
- https://doi.org//10.1016/j.urolonc.2018.12.011
- https://doi.org//10.1038/s41598-020-70896-0
- https://doi.org//10.1096/fj.202002168RR
- https://doi.org//10.3322/caac.21654
- https://doi.org//10.1096/fj.06-5885fje
- https://doi.org//10.1056/NEJMoa1001283
- https://doi.org//10.1038/srep37095
- https://doi.org//10.3390/ijms22189850
- https://doi.org//10.1093/humrep/deh609
- https://doi.org//10.1093/humrep/dex357
- https://doi.org//10.1038/nprot.2006.179
- https://doi.org//10.1007/s12064-012-0162-3
- https://doi.org//10.1111/j.1749-0774.2002.tb00105.x
- https://doi.org//10.1210/mend.11.5.9928
- https://doi.org//10.1038/sj.onc.1204632
- https://doi.org//10.1016/j.bbrc.2010.02.112
- https://doi.org//10.1016/j.bbrc.2012.04.104
- https://doi.org//10.1016/j.celrep.2017.11.023
- https://doi.org//10.1016/j.cell.2011.02.013
- https://doi.org//10.1038/ng.2764
- https://doi.org//10.1038/s41598-017-01617-3
- https://doi.org//10.1002/ijgo.12612
- https://doi.org//10.23736/s0026-4784.18.04272-7
- https://doi.org//10.1016/j.mce.2016.08.005
- https://doi.org//10.3389/fphar.2020.602593
- https://doi.org//10.1016/j.bmcl.2014.03.018
- https://doi.org//10.1101/cshperspect.a006098
- https://doi.org//10.1038/s41586-018-0409-3
- https://doi.org//10.2337/diabetes.54.6.1726
- https://doi.org//10.7554/eLife.07485
- https://doi.org//10.3390/cells8030238
- https://doi.org//10.1242/jcs.145854
- https://doi.org//10.1074/jbc.M115.638924
- https://doi.org//10.1194/jlr.M025403
- https://doi.org//10.1038/s41416-019-0451-4
- https://doi.org//10.3389/fonc.2020.605154
- https://doi.org//10.1210/jcem-70-2-421
- https://doi.org//10.1530/rep.0.1210835
- https://doi.org//10.1097/IGC.0000000000000384
- https://doi.org//10.1158/1541-7786.Mcr-12-0219
- https://doi.org//10.1002/1873-3468.14017
- https://doi.org//10.1074/jbc.M402121200
- https://doi.org//10.3892/ijmm.2017.3288
- https://doi.org//10.1038/s41598-017-14676-3
- https://doi.org//10.1038/s41591-020-1040-z