Donatella Aielloa, Giuseppina Cacciatorea, Vincenzo Letteraa, Marco Malavoltab, Serena Marcozzib, Giorgia Bigossib, Angela Amoresanoc, Anna Napolia

aDipartimento di Chimica e Tecnologie Chimiche, Università della Calabria, Arcavacata di Rende, 87036, Cosenza, Italy;
bAdvanced Technology Center for Aging Research and Geriatric Mouse Clinic, IRCCS INRCA, 60121, Ancona, Italy;
cDipartimento di Scienze Chimiche, Università Federico II di Napoli, Via Cinthia 26, 80126, Napoli, Italy.

In recent years, human longevity research has emerged as a topic of great interest related to improving health and lifespan. However, studying the causes of aging is difficult as the biological process is multifunctional and involves molecular, cellular and physiological changes [1]. In this context, proteomics has emerged as a powerful approach to investigate protein changes during aging, identify biomarkers to assess the state of biological aging and predict age-related diseases [2]. This study aims to discover potential biomarkers through differential proteomics analysis of red blood cells from groups of individuals across different ages, in order to qualitatively and quantitatively characterize the differences and similarities of each group at molecular level. Initial experiments concerning extraction methods and analysis were conducted on Mus musculus, as a model organism. Blood samples were obtained from mice at three life stages: young (6 months), middle-aged (12 months), and old (30 months). The membrane protein content was then extracted and separated. To identify potential biomarkers associated with membrane proteins, an untargeted LC-MS/MS proteomics approach was employed. The data obtained were processed using MaxQuant software and the result was further analyzed with STRING to integrate all known and predicted interactions between proteins that were either upregulated or downregulated. Many of the differentially expressed proteins identified play a key role in pathophysiological processes, such as Apolipoprotein A-1, Alpha-actinin-1, Complement factor H, Eukaryotic translation initiation factor 5A-1. This study reveals potential biomarkers of aging in blood cells and explains how they relate to the immune system or metabolic pathways. The data obtained will be used to develop a comprehensive model of the molecular mechanisms underlying human longevity, either as self-consistent indicators or integrated with data from other omics approaches.

Acknowledgements: this work was financially supported by the Research Programm “ Age-it- Ageing well in an ageing society”, Spoke 2 - Project code PE0000015, CUP B83C22004800006, founded by the PNRR, Missione 4 “Istruzione e Ricerca”.

Keywords: Aging, Healthspan, Proteomics, Biomarkers.

References

1. Shen X., Wang C., Zhou X. et al., Nat Aging, (2024). DOI: 10.1038/s43587-024-00692-2 

2. Johnson A., Shokhirev M. N., et al., Ageing Res. Rev., 60 (2020). DOI: 10.1016/j.arr.2020.101070