Adaptive thermogenesis reprograms the behavioural and metabolic responses to protein restriction for successful amino acid homeostasis

Tsang, A.; Benoit, S.; Lockhart, S.; Parish, A.; Virtue, S.; da Silva Paes, L.; Wong, X. L.; Rimmington, D.; Vidal-Puig, A.; O'Rahilly, S.; Coll, A. P.; Pirro, V.; Koulman, A.; Blouet, C.

doi:10.1101/2025.07.30.667623

bioRxiv · 10.1101/2025.07.30.667623

Adaptive thermogenesis reprograms the behavioural and metabolic responses to protein restriction for successful amino acid homeostasis

Tsang, A.·Benoit, S.·Lockhart, S.·Parish, A.·Virtue, S.·da Silva Paes, L.·Wong, X. L.·Rimmington, D.·Vidal-Puig, A.·O'Rahilly, S.·Coll, A. P.·Pirro, V.·Koulman, A.·Blouet, C.

View original source ↗Export metadata ↓

Abstract

Reduced protein intake is proposed to contribute to the obesity epidemic, but existing murine models of protein restriction do not promote obesity, limiting insight into underlying mechanisms. Here we show that mice self-select a consistent daily protein intake, independent of energy needs. Under mild protein restriction (7-10% protein), mice exhibit hyperphagia and increased adiposity. This hyperphagic response is blunted at thermoneutrality, leading to loss of lean mass and body weight. Protein-restricted mice also fail to exhibit protein preference at thermoneutrality, despite elevated circulating FGF21 levels. Circulating levels of essential amino acids (AAs) are tightly regulated during protein restriction at 22{degrees}C, but this regulation is lost at 28{degrees}C. Metabolomic and transcriptional analyses revealed a role for hepatic and brown fat AA-derived acylcarnitines and N-acetyl AAs in buffering free AA levels for the maintenance of AA homeostasis at 22{degrees}C, but these pathways are blunted at thermoneutrality. Thus, the behavioural and metabolic adaptations to protein restriction rely on coordinated peripheral and central mechanisms, modulated by ambient temperature. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=63 SRC="FIGDIR/small/667623v1_ufig1.gif" ALT="Figure 1"> View larger version (19K): org.highwire.dtl.DTLVardef@1d1a9e1org.highwire.dtl.DTLVardef@1e7f3b5org.highwire.dtl.DTLVardef@131276borg.highwire.dtl.DTLVardef@130757_HPS_FORMAT_FIGEXP M_FIG C_FIG

Explore related subjects

physiology Preprint

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Tsang, A., Benoit, S., Lockhart, S., Parish, A., Virtue, S., da Silva Paes, L., Wong, X. L., Rimmington, D., Vidal-Puig, A., O'Rahilly, S., Coll, A. P., Pirro, V., Koulman, A., Blouet, C.. 2025-08-02. Adaptive thermogenesis reprograms the behavioural and metabolic responses to protein restriction for successful amino acid homeostasis. https://doi.org/10.1101/2025.07.30.667623

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

28S-rsRNA facilitates PASMC proliferation through PKM2 β-hydroxybutyrylation

Ribosomal RNA-derived small RNAs (rsRNAs) are nucleic acid fragments originating from rRNA, the functions and roles of which in diseases remain incompletely understood. We identified 28S rsRNA-CTC_L20, an rsRNA whose expression is upregulated in hypoxic pulmonary artery smooth muscle cells (PASMCs) and in lung tissues of pulmonary hypertension (PH) models. In hypoxic cells, 28S rsRNA-CTC_L20 promotes PASMC proliferation, and its knockdown mitigates SuHx-induced pulmonary hypertension and vascular remodeling. Mechanistically, 28S rsRNA-CTC_L20 modulates {beta}-hydroxybutyrate (BHB) production by maintaining the stability of ketogenic enzyme mRNAs through its "CTGNNGG" motif. Elevated levels of BHB enhance the {beta}-hydroxybutyrylation modification at K166 of pyruvate kinase M2 (PKM2), increasing its activity and driving glycolysis to sustain proliferation. These results link ketone body metabolism to glycolysis in hypoxic cells and define an rsRNA-BHB-PKM2 {beta}-hydroxybutyrylation axis in PH. They also point to 28S rsRNA-CTC_L20 as a potential target for treating vascular remodeling.

physiology↗

bioRxiv2026

Selective activation of estrogen receptors modulates electrical activity and insulin secretion in human EndoC-βH1 cells

Estrogen receptors (ERs), ER and ER{beta} have important roles in the physiology of the endocrine pancreas in animal models, but the effects of ER activation on insulin-secreting human cells are still poorly understood. Here, we investigated the role of ER and ER{beta} as modulators of electrical activity and insulin secretion in the insulin-secreting human cell line EndoC-{beta}H1. Immunocytochemical analysis revealed a widespread cellular distribution of ER, while ER{beta} was predominantly localized outside the nucleus. We observed that both estradiol (E2) and the ER{beta} agonist diarylpropionitrile (DPN) led to a decrease in insulin secretion in the absence of stimulatory glucose concentrations, without affecting insulin content. Patch-clamp experiments revealed that a 48-hour treatment with either the ER agonist PPT or DPN resulted in depolarization of the resting membrane potential, increased action potential firing frequency, and altered the action potential waveform in EndoC-{beta}H1 cells. PPT increased voltage-dependent Ca2+ channel currents, while DPN reduced K+ and Na+ currents. These results suggest that activation of ER and ER{beta} modulates beta-cell electrical activity through distinct pathways involving ion channels. The effects of E2, PPT, and DPN were not caused by altered expression levels of ion channel genes after 48 hours of treatment. This suggests that these receptors act extranuclearly without affecting gene expression. Together, our findings demonstrate that selective activation of ER and ER{beta} modulates beta-cell excitability through distinct electrophysiological mechanisms and that E2 and DPN reduce basal insulin secretion in human EndoC-{beta}H1 cells. These results support ER and ER{beta} as potential targets for modulating human pancreatic beta-cell function.

physiology↗

bioRxiv2026

Body weight modifies the efficacy of lifespan-extending interventions in ad libitum-fed mice

Geroprotective interventions are often assessed by extension of lifespan in ad libitum-fed mice. Increased lifespan could therefore reflect rescue of adverse phenotypes of unhealthy controls. We analysed body weight and survival for 33,780 mice in the Interventions Testing Program (ITP) to test whether treatment effects varied with body weight. In males, each added standard deviation of attained adult weight made treatment 6% more favourable (HR 0.94), more apparent for interventions that lowered body weight (HR 0.90). Median lifespan increase rose from 6.2% in mice in the lightest quartile to 20.2% in the heaviest pre-dosing weight quartile among those experiments, mainly because control survival declined more steeply with weight than that of the intervention group. The effect of body weight was consistent across 13 of 14 compounds and reflected weight attained during adulthood rather than in early life. Weight-dependence of intervention effects was weak in ITP females, possibly because median survival of controls exceeded that of every sampled non-ITP female control group. Amelioration of weight-associated risk is therefore implicated as a major contributor to lifespan increases relative to heavy, short-lived controls. The geroprotective effects of these interventions may thus be less apparent in metabolically healthier mice or humans.

physiology↗