PROJECT SUMMARY/ABSTRACT
Premature ovarian insufficiency (POI) is characterized by a decline in ovarian function resulting in infertility and
reduced ovarian hormone production. Globally, the rate of POI is approximately 3.5% resulting in
~138,250,000 women experiencing subfertility or infertility. Consequently, fertility preservation and restoration
are a priority among patients with POI and individuals who desire children later in their reproductive lifespan.
However, many limitations to the current fertility preservation and restoration options result in short hormonal
functionality of transplanted ovarian tissue, low live birth rates, and low numbers of good-quality oocytes for in
vitro maturation. Therefore, the development of novel technologies and methodologies for the advancement of
reproductive science is imperative. Our main goal through this work is to elucidate matrisome-dependent
mechanisms that contribute to folliculogenesis while also designing a defined biomaterial that improves follicle
growth and oocyte quality in vitro. We will accomplish this through the interrogation of Elastin Microfibril
Interface-located protein 1’s (EMILIN1) role in folliculogenesis (Aim 1) and by engineering a biomaterial that
increases follicle growth and oocyte quality (Aim 2). Our preliminary studies demonstrate our ability to
investigate matrisome-dependent cues in vitro using decellularized extracellular matrix (dECM) hydrogels
derived from bovine ovarian tissue. We have shown that hydrogels with a high quantity of EMILIN1 increase
murine follicle diameter in vitro compared to hydrogels with a low quantity of EMILIN1. This effect can be
rescued with the addition of rhEMILIN1. Importantly, this effect can only be observed when follicles are cultured
in dECM and EMILIN1 had no effect on growth when added to a biologically inert hydrogel, alginate. Therefore,
we will elucidate the mechanism by which EMILIN1 increases follicle growth by performing bulk-RNA
sequencing on murine follicles grown in the presence or absence of EMILIN1 in dECM and interrogating the
signaling pathways associated with EMILIN1. Then, we will engineer a hydrogel of defined matrisome
composition to recapitulate the follicle growth dynamics observed in dECM hydrogels with an abundance of
EMILIN1. To do this, we will perform a co-immunoprecipitation (co-IP) and subsequent unbiased proteomics to
define matrisome proteins that EMILIN1 associates with. We will then use this list of proteins to design an ECM
hydrogel for ovarian follicle culture that increases follicle growth and oocyte quality via an EMILIN1-driven
mechanism. Taken together, these aims will expand our understanding of the role of EMILIN1 in
folliculogenesis, advise future work in designing advanced biomaterials for follicle culture, and lead to future
fertility preservation and restoration options. Finally, this research will be influential to the greater scientific
community by expanding our technical repertoire for studying matrisome-dependent biochemical cues within
other organ systems.
