São Paulo Researchers in International Collaboration Université Laval/Quebec-Canada and Federal University of ABC São Paulo-Brazil

• Grant number: 24/17019-0
• Support Opportunities: Regular Research Grants
• Start date: February 01, 2025
• End date: January 31, 2027
• Field of knowledge: Agronomical Sciences - Veterinary Medicine - Animal Reproduction
• Mobility Program: SPRINT - Projetos de pesquisa - Mobilidade
• Principal Investigator: Marcella Pecora Milazzotto
• Grantee: Marcella Pecora Milazzotto
• Principal researcher abroad: Marc Andre Sirard
• Institution abroad: Université Laval, Canada
• Host Institution: Centro de Ciências Naturais e Humanas (CCNH). Universidade Federal do ABC (UFABC). Ministério da Educação (Brasil). Santo André , SP, Brazil
• Associated research grant: 23/15734-1 - The Role of S-Adenosylmethionine Sensor in the Metabolic and Epigenetic Profile of the Pre-Implantation Embryo, AP.R

Abstract

During early development, the pre-implantation embryo undergoes an intense process of metabolic, molecular, and epigenetic reprogramming, which must be adequately orchestrated to support the events that will culminate in the generation of a new individual. Understanding these mechanisms is of fundamental importance for the development of increasingly efficient assisted reproduction techniques. A central event in early development is the reprogramming of nuclear and mitochondrial DNA methylation, which, among other factors, depends on the appropriate metabolic profile of the cell. Recent studies have shown that the mechanistic target of rapamycin complex 1 (mTORC1) acts as a common element controlling various mechanisms that directly interfere with the DNA methylation profile. In particular, one of the control mechanisms of mTORC1 is the sensor for S-adenosylmethionine (SAM- the main methyl group donor for DNA methylation), SAMTOR. In the presence of methionine, SAM is synthesized and binds to SAMTOR, leading to the activation of mTORC1, which in turn activates its downstream targets, including DNA methyltransferase 1 (DNMT1) and hypoxia-inducible factor 1-alpha (HIF-1¿), which, simplistically, result in an increase in DNA methylation and the activity of the glycolytic pathway, respectively. Thus, in this work, we propose to understand the role of SAMTOR in metabolic and epigenetic control during embryonic development. For this purpose, SAMTOR knockdown will be performed starting from the zygote stage, and the produced embryos will be evaluated for the quantity and localization of mTOR, HIF-1¿, PDK1, and DNMT1, mitochondrial membrane potential, as well as glycolytic pathway transcripts and the establishment of DNA methylation. Additionally, the global nuclear DNA methylation profile will be analyzed throughout development, as well as the nuclear and mitochondrial methylome profiles under SAMTOR inhibition. It is expected that these results will contribute to the understanding of the regulatory mechanisms of DNA demethylation and de novo methylation processes in embryos and the role of SAMTOR and mTORC1 in this process. (AU)