Sex-specific regulation of metabolic health and vertebrate lifespan by AMP biosynthesis

Abstract

The loss of energy homeostasis seen during aging, is causally linked to multiple age-related pathologies. The AMP-activated protein kinase (AMPK) directly senses cellular energy levels, which are reflected in the ratio between AMP:ATP. However, the genetic regulation of vertebrate aging by the AMPK pathway remains poorly understood. Here, we manipulate ATP production by mutating APRT, a key enzyme in AMP biosynthesis, and extend vertebrate lifespan in a male-specific manner. Using a multi-omics approach, we demonstrate that the APRT mutation restores metabolic plasticity, and identify a distinct transcriptional signature linking mitochondria with the sex-related differences in longevity. Accordingly, APRT mutant cells display a reduction in mitochondrial functions and ATP levels, and an increase in AMPK activity, resembling a persistent state of energy starvation. In-vivo, a fasting-like response was observed exclusively in male mutants, including resistance to a high-fat diet. Finally, intermittent fasting eliminated the longevity benefits mediated by the APRT mutation in males. Together, these data identify AMP biosynthesis as a sex-specific mediator of vertebrate longevity and metabolic health.

Refeeding-associated AMPKγ1complex activity is a hallmark of health and longevity

“…This could indicate that AMPK γ2 function is physiologically required exclusively upon fasting and explain why, unlike AMPK γ1 , chronic activation of AMPK γ2 leads to metabolic dysfunctions in normally fed mice or humans 45,46 . Astre et al showed that mutation in the APRT gene, a key enzyme in AMP biosynthesis, mimics a caloric-restriction effect, inducing lifespan extension in killifish males 47 . This effect was associated with higher expression of the γ2 subunit, possibly indicating that higher activation of AMPK γ2 is beneficial under energetic stress conditions and implies that various dietary interventions might represent a concrete strategy to counter the pathological effect of AMPK γ2 hyperactivity in humans.…”

A scalable platform for functional interrogation of peptide hormones in fish

“…We have recently demonstrated that the killifish model could be used to identify novel regulators of aging, which involve systemic modulation of metabolism 14 . Therefore, beyond our current application, it would be exciting to expand this platform to investigate systemic factors as modifiers of health and disease.…”

“…Tissues samples were processed as described previously [12][13][14][15][48][49][50][51][52][53][54][55][56][57][58][59] . Briefly, fish were euthanized with 500 mg/l tricaine (MS222, #A5040, Sigma).…”

“…The hypothalamic-pituitary-somatic (HPS) axis includes the secretion of growth hormone (GH) from the pituitary gland, and the consequent stimulation of insulin-like growth factor 1 (IGF-1) production in peripheral tissues (primarily the liver). We applied our recently developed CRISPR/Cas9 genome editing protocols [12][13][14][15] to perturb the HPS axis in the turquoise killifish by targeting exon 2 of the killifish gh1 (Figure 1b, top). Crossing F0 founders with wild-type fish allowed us to identify a 4bp deletion in F1 fish that could cause a frame-shift.…”

Rapid and precise genome engineering in a naturally short-lived vertebrate

“…Tools to advance genetic interrogation of the killifish have been developed, including a sequenced genome (Reichwald et al, 2015; Valenzano et al, 2015) and Tol2 transgenesis (Allard et al, 2013; Hartmann and Englert, 2012; Valenzano et al, 2011), as well as CRISPR/Cas9-mediated knock-out (Harel et al, 2015) and CRISPR/Cas13-mediated knock-down (Kushawah et al, 2020). This genetic toolkit has enabled discoveries about the mechanisms of aging (Astre et al, 2022a; Bradshaw et al, 2022; Chen et al, 2022; Harel et al, 2022; Louka et al, 2022; Matsui et al, 2019; Smith et al, 2017; Van Houcke et al, 2021b), regeneration (Vanhunsel et al, 2022a; Vanhunsel et al, 2021; Vanhunsel et al, 2022b; Wang et al, 2020), evolution (Cui et al, 2019; Sahm et al, 2017; Singh et al, 2021; Willemsen et al, 2020), development (Abitua et al, 2021; Dolfi et al, 2019), and ‘suspended animation’ (Hu et al, 2020; Singh et al, 2021).…”