Transfection of extracellular vesicles from follicular fluid with specific microRNAs: a form of miRNA delivery to bovine cumulus-oocyte complex cells

• Grant number: 22/09461-0
• Support Opportunities: Scholarships in Brazil - Master
• Start date: March 01, 2023
• End date: August 26, 2025
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Principal Investigator: Juliano Coelho da Silveira
• Grantee: Camila Azzolin de Souza
• Host Institution: Faculdade de Zootecnia e Engenharia de Alimentos (FZEA). Universidade de São Paulo (USP). Pirassununga , SP, Brazil
• Associated research grant: 21/06645-0 - Extracellular vesicles as a platform for diagnostic and manipulation of the in vitro embryo production system: the next generation in animal reproduction, AP.JP2
• Associated scholarship(s): 24/07395-5 - Supplementation with specific miRNAs during early embryo development: A tool for improving in vitro embryo production, BE.EP.MS

Abstract

Reproductive biotechnologies such as in vitro embryo production accelerate genetic improvement, but its success is directly linked to the ability to mimic the in vivo environment. Among the events that occur during follicular maturation and oocyte growth is the intercellular communication in the follicular environment and consequently the transfer of biomolecules between somatic cells and between somatic and gametic cells. The transference of these molecules is facilitated by the follicular fluid present in the follicular antrum. Follicular fluid provides an important microenvironment for follicular growth and oocyte maturation, as it contains, among other components, extracellular vesicles (EVs). EVs are nanoparticles that contain various biological molecules, such as microRNAs, that can modulate the function of recipient cells. Therefore, the hypothesis of the present study is that extracellular vesicles from the follicular fluid can be loaded with specific miRNAs and are able to deliver these miRNAs to cumulus cells in vitro. Thus, the objectives are: 1) to isolate and transfect extracellular vesicles from the follicular fluid with miRNAs, 2) to verify the transfection efficiency of miRNAs, 3) to determine the delivery efficiency of miRNAs, 4) to determine the molecular effects of bovine COC supplementation. with extracellular vesicles containing miRNAs of interest and 5) to evaluate the rates of maturation and embryo production of COCs supplemented with extracellular vesicles containing miRNAs of interest. We hope to demonstrate the successful transfection of extracellular vesicles with specific miRNAs as well as their delivery to cumulus cells. The results of our study are of fundamental importance to fill gaps in reproductive biology, in the same way that it presents itself as a potential tool to increase in vitro embryo production rates.