Understanding The Role Of FSHR Oligomerization And Trafficking In Transducing Age-dependent Changes In FSH Glycoforms

Project Summary/Abstract Although ovarian aging is a natural physiological process, cessation of ovarian function at midlife increases susceptibility to the development of co-morbidities, such as osteoporosis, which decrease quality of life and increases healthcare burden. There is currently no intervention for preventing/delaying this, highlighting an

unmet need for novel approaches, e.g. extending ovarian lifespan, to tackle this. Key mediators of ovarian function are follicle stimulating hormone (FSH) and its receptor (FSHR). FSH is a glycoprotein hormone, with two predominant glycoforms identified; partially glycosylated FSH (FSH18/21) and fully glycosylated FSH

(FSH24). FSH glycoforms have distinct functional properties, with FSH18/21 displaying faster binding kinetics and more potent and distinct signal pathway activation than FSH24. Interestingly, age-related changes in human pituitary extracts have been reported, with FSH18/21 predominant in women of reproductive prime and FSH24

predominant in peri-menopausal/menopausal women. The increase in less potent FSH24 coincides with increasing ovarian resistance to FSH reported during peri-menopause. Yet, how these functional differences in FSH modulate the actions of FSHR within the aging ovary remain unknown and will be explored by

this project. Increasingly important ways that G protein-coupled receptors direct signal and functional diversity is via oligomerization (association of receptor protomers into dimers and oligomers), and endosomal trafficking. Our recent work suggests that FSH18/21 and FSH24 distinctly modulate FSHR oligomerization, correlating with

differences in FSH glycoform-dependent cAMP production. Our preliminary data suggest that FSH glycoforms may route FSHR to different endosomal compartments, with cAMP production dependent on FSHR internalization. This project aims to determine how FSH glycoforms modulate FSHR oligomerization and trafficking in young and aging ovaries, and functional impact thereof. The working hypothesis is FSH glycoforms

mediate distinct FSHR oligomerization signatures and FSHR trafficking within the young and aging ovary, with defined functional consequences. Using a novel N terminal tagged FLAG-FSHR knock in mouse, high resolution imaging and multiomics ATACseq and RNAseq approaches, Aim 1 will determine how FSHR oligomerization

and signaling is modulated in granulosa cells during folliculogenesis and aging, and how disrupting FSHR oligomerization impacts FSH glycoform function. Aim 2 will determine how FSH glycoforms direct FSHR endosomal routing and role of these compartment in signal activation and downstream functions. Successful

completion of these aims will elucidate how unique ovarian FSHR oligomerization and trafficking signatures direct signal pathway activation and transcriptomic and functional outputs by FSH glycoforms, during aging. This is the first step in the discovery pipeline to identifying novel strategies to improve

ovarian function and health in aging women.