A renal clearable fluorogenic probe for in vivo β-galactosidase activity detection during aging and senolysis

Sara Rojas-Vázquez, Beatriz Lozano-Torres, Alba García-Fernández, Irene Galiana, Ana Perez-Villalba, Pablo Martí-Rodrigo, M. José Palop, Marcia Domínguez, Mar Orzáez, Félix Sancenón, Juan F. Blandez, Isabel Fariñas () and Ramón Martínez-Máñez ()
Additional contact information
Sara Rojas-Vázquez: Universitat Politècnica de València-Universitat de València
Beatriz Lozano-Torres: Universitat Politècnica de València-Universitat de València
Alba García-Fernández: Universitat Politècnica de València-Universitat de València
Irene Galiana: Universitat Politècnica de València-Universitat de València
Ana Perez-Villalba: Universidad Católica de Valencia
Pablo Martí-Rodrigo: Universitat de València
M. José Palop: Universitat de València
Marcia Domínguez: Universitat Politècnica de València-Universitat de València
Mar Orzáez: Universitat Politècnica de València, Centro de Investigación Príncipe Felipe
Félix Sancenón: Universitat Politècnica de València-Universitat de València
Juan F. Blandez: Universitat Politècnica de València-Universitat de València
Isabel Fariñas: Universitat de València
Ramón Martínez-Máñez: Universitat Politècnica de València-Universitat de València

Nature Communications, 2024, vol. 15, issue 1, 1-16

Abstract: Abstract Accumulation of senescent cells with age leads to tissue dysfunction and related diseases. Their detection in vivo still constitutes a challenge in aging research. We describe the generation of a fluorogenic probe (sulfonic-Cy7Gal) based on a galactose derivative, to serve as substrate for β-galactosidase, conjugated to a Cy7 fluorophore modified with sulfonic groups to enhance its ability to diffuse. When administered to male or female mice, β-galactosidase cleaves the O-glycosidic bond, releasing the fluorophore that is ultimately excreted by the kidneys and can be measured in urine. The intensity of the recovered fluorophore reliably reflects an experimentally controlled load of cellular senescence and correlates with age-associated anxiety during aging and senolytic treatment. Interestingly, our findings with the probe indicate that the effects of senolysis are temporary if the treatment is discontinued. Our strategy may serve as a basis for developing fluorogenic platforms designed for easy longitudinal monitoring of enzymatic activities in biofluids.

Date: 2024
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DOI: 10.1038/s41467-024-44903-1

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