Sarcolipin-associated transcription factors: New road to thermogenesis

• Grant number: 24/05514-7
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• Start date: January 01, 2025
• End date: December 31, 2027
• Field of knowledge: Biological Sciences - Biochemistry - Molecular Biology
• Principal Investigator: Leonardo dos Reis Silveira
• Grantee: Carolina Martins Lazaro
• Host Institution: Instituto de Biologia (IB). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Associated research grant: 23/08207-5 - New road to thermogenesis: targeting nuclear receptor Nr2f6 in skeletal muscle, AP.TEM

Abstract

Obesity poses a significant global health challenge, prompting the need for innovative therapeutic approaches. One promising avenue involves unraveling the molecular mechanisms underlying thermogenesis, particularly in skeletal muscle. Sarcolipin (SLN), a regulatory of sarcoplasmic reticulum calcium ATPase (SERCA) binds to SERCA in a Ca2+-sensitive manner and its binding can lead to uncoupling of SERCA pump activity. During this activity, SLN promotes futile cycling of the pump, increases heat generation, and plays an important role in muscle thermogenesis. Despite its importance, the transcription factors (TFs) orchestrating SLN expression and their role in thermogenesis regulation remain largely unexplored. This project aims to address this critical gap by elucidating the TFs responsible for enhanced SLN expression and their subsequent impact on thermogenesis in skeletal muscle cells. Utilizing established methodologies, including lentiviral knockdown constructs and reporter assays, we will induce SLN expression in primary muscle cells from mice using lentiviral constructs. Then, through immunoprecipitation (enChIP) and mass spectrometry (MS) techniques, we will identify TFs binding to SLN DNA sequence. Due SLN's regulatory effect over SERCA, we hypothesize that specific TFs of SLN could increase SERCA uncoupled activity. This effect has been associated with greater ATP consumption and futile energy expenditure. Furthermore, enChIP will help us to map interactions between TFs and SLN regulatory regions, shedding light on the intricate transcriptional control mechanisms underlying thermogenic activity. Functional validation of candidate TFs will be achieved through metabolic analyses, such as respirometry, SERCA activity, Ca²¿ flux assays, and transcriptional profiling of metabolic genes linked to thermogenesis as well as fatty acid and glucose metabolism. This interdisciplinary approach integrates molecular biology, bioinformatics, and metabolic physiology to unravel the complex interplay between TFs, SLN expression and SERCA uncoupling on thermogenic pathways in skeletal muscle. By identifying novel TFs involved in SLN expression, our research holds the potential to uncover therapeutic targets for combating obesity and metabolic disorders.