Mammals display wide range of variation in their lifespan. Lifespan is generally 29 correlated to body size, but outliers such as human and the naked mole-rat 30 (NMR, Heterocephalus glaber) exist. Investigating the molecular networks that 31 distinguish long-from short-lived species has proven useful to identify 32 determinants of longevity. Here, we compared the liver of long-lived NMRs and 33 the phylogenetically closely related, shorter-lived, guinea pigs (GP, Cavia 34 porcellus) using an integrated transcriptomic and proteomic approach. We found 35 that NMRs express substantially higher levels (up to 30 fold) of a restricted 36 number of longevity-associated proteins that confer enhanced buffering against 37 oxidative stress. Moreover, NMR livers display a unique expression pattern of 38 mitochondrial proteins that result in distinct metabolic features of their 39 mitochondria. For instance, we observed a generally reduced respiration rate 40 associated with lower protein levels of respiratory chain components, 41 particularly complex I, and increased capacity to utilize fatty acids. Interestingly, 42 the same molecular networks are affected during aging in both NMR and 43 humans, supporting a direct link to the extraordinary longevity of both species. 44Finally, we used our analysis to identify novel longevity pathways, and validated 45 one of them experimentally in the phylogenetically distantly related nematode C. 46 elegans. 47 48 . CC-BY 4.0 International license not peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was . http://dx.doi.org/10.1101/220343 doi: bioRxiv preprint first posted online Nov. 17, 2017; Heinze, Bens, Calzia, et al.
2Introduction: 49 50Among mammals lifespan generally correlates with other life-history parameters 51 such as gestation period and body mass [1]. In this perspective, a subterranean 52 rodent, the naked mole-rat (Heterocephalus glaber, NMR), and humans represent 53 two species outliers by having an exceptionally long lifespan relatively to their 54 body mass. NMRs are eusocial animals that live in colonies where only a 55 subgroup of animals is devoted to reproduction (usually a queen and one male 56 called pasha) [2]. NMRs exhibit other exceptional traits including lifelong 57 fertility, resistance to infection, high regenerative capacity, resistance to cancer 58 and diabetes, reviewed in [3,4]. For these reasons, NMRs have drawn attention of 59 multiple studies aimed at identifying the molecular mechanisms behind their 60 extreme longevity and resistance to age-related diseases. Comparative genome 61 analysis has revealed positively selected genes in NMR [5,6], and RNA-seq 62 analysis revealed minimal changes in gene expression during aging [5,7], 63 supporting the view of enhanced maintenance of homeostasis in NMRs at the 64 molecular level. NMRs possess enhanced protein stability and increased 65 proteasomal activity [8,9], negligible levels of cellular senescence [10], over-66 activation of pathways...