Blocking estrogen synthesis via aromatase inhibition helps prevent the recurrence of estrogen receptor-positive breast cancer but also results in cognitive, sleep, and thermoregulatory disturbances. These side effects diminish quality of life and contribute to treatment nonadherence in a large proportion of patients. 10β,17β-Dihydroxyestra-1,4-dien-3-one (DHED) is a brain-selective prodrug that, in rodent models, converts to 17β-estradiol (E 2) selectively in the brain without affecting the periphery. We investigated whether DHED could prevent side effects associated with the aromatase inhibitor (AI) letrozole in a primate model of aging. Chronic oral treatment with DHED in letrozole-treated male and female marmosets led to a robust increase in E 2 levels across brain regions without affecting estrogen levels at the periphery. In addition, DHED treatment (1) improved memory at short delays and prevented letrozole-induced cognitive slowing in a hippocampal-dependent memory task, (2) normalized hippocampal neuronal membrane potential and excitability, and (3) reduced sleep fragmentation. However, DHED treatment had opposite effects on thermoregulation in males and females, necessitating additional research in this area. Overall, the results suggest that DHED, which lacks estrogenic effects in peripheral tissues, could be a safe and effective novel hormonal therapy for improving quality of life in breast cancer patients treated with AIs.

Menopausal symptoms of sleep disturbances, cognitive deficits, and hot flashes are understudied, in part due to the lack of animal models in which they co-occur. Common marmosets (Callithrix jacchus) are valuable nonhuman primates for studying these symptoms, and we examined changes in cognition (reversal learning), sleep (48 h/wk of sleep recorded by telemetry), and thermoregulation (nose temperature in response to mild external warming) in middle-aged, surgically-induced menopausal marmosets studied at baseline, during 3-week phases of ethinyl estradiol (EE2, 4 μg/kg/day, p.o.) treatment and after EE2 withdrawal. We also assessed a brain-selective hormonal therapy devoid of estrogenic effects in peripheral tissues on the same measures (cognition, sleep, thermoregulation) after treatment with the estrogen prodrug 10β,17β-dihydroxyestra-1,4-dien-3-one (DHED, 100 μg/kg/day, p.o) and DHED withdrawal. Reversal learning performance was improved with EE2 or DHED treatment relative to phases without hormone administration, as indicated by a faster reversal of the stimulus/reward contingencies. Both EE2 and DHED increased non-REM sleep and reduced nighttime awakenings relative to baseline, but to the detriment of REM sleep which was highest at baseline. Nasal temperature in response to mild external warming was highest, and overnight core body temperature lowest, in the DHED treatment phase compared to both the EE2 and baseline phases. These results suggest that low dose estradiol, delivered either peripherally or centrally via DHED, benefits selective aspects of cognition and sleep in a marmoset menopause model. DHED appears a promising therapeutic candidate for alleviating the cognitive and sleep disruptions associated with estrogen deficiency in primates.

Nonhuman primates are arguably the best models for Alzheimer’s disease (AD), due to close similarities with humans in physiology and behavior, brain structure and function, and aging patterns. The common marmoset (Callithrix jacchus) is particularly valuable because it has the shortest lifespan of all anthropoids (10-12 years), exhibits sex differences in age-related cognitive decline and develops AD-like pathology with age. In the present study, we investigated blood-based biomarkers in male and female marmosets with cognitive and neuropathological assessments. We analyzed the concentrations of several AD biomarkers from blood samples of male and female marmosets (n = 13, age range 6.8-9.36 years) characterized for age-related cognitive decline and neuropathology. AD biomarker concentrations were measured using the commercially available S-PLEX neurology panel 1 kit (NHP) from Mesoscale Discovery. The kit, designed for GFAP, NfL, and tau detection, employs electrochemiluminescence to discern femtogram concentrations. Similar to ELISA, the emitted light intensity in S-PLEX assays directly correlates with analyte amounts in the sample. Log-transformed biomarker data were subjected to analyses of variance, with Sex and Cognitive Status as factors, and Age as a covariate. LogGFAP concentrations were significantly higher in cognitively impaired (F (1, 8) = 15.50, p < .01) than unimpaired animals. Other biomarkers did not differ significantly based on cognitive status. All biomarkers exhibited non-significant Sex effects (all ps < .10), suggesting higher levels in males. A non-significant Sex x Cognitive Status interaction for LogGFAP (F (1, 8) = 4.12, p < .08) indicated higher levels in males among the impaired group. Although we found no significant association between blood biomarkers and neuropathological data, substantial correlations were noted within the impaired group. Analyses with a larger sample size are in progress. Alterations in biomarker concentrations serve as early indicators of cognitive decline, preceding AD diagnosis. Elevated blood GFAP, indicative of inflammation, differentiated impaired and non-impaired aged marmosets, with sex influencing biomarker levels, predominantly higher in males. The absence of significant correlations between blood biomarkers and neuropathology necessitates further validation with a larger subject cohort. Supported by NIH R01 AG046266 and SAGA 23

Nonhuman primate models of human aging are necessary to understand functional decline and identify vulnerabilities to aging pathologies. Longitudinal studies are critical and common marmosets ( Callithrix jacchus ) are ideal as they have a short lifespan (∼12 yr) and naturally develop neuropathology relevant to Alzheimer’s disease (AD). We studied 28 marmosets (14 females) from midlife (∼5 yr old) to old age across four years. Simple discrimination and reversal learning was used to evaluate cognitive performance. Additional yearly tests included motor, stress reactivity, and social behavior assessments. A subset of animals (n = 17, 9 females) were studied for AD-relevant neuropathology in post-mortem tissue of the dorsal lateral prefrontal cortex (dlPFC) and CA1 of the hippocampus. Multilevel models revealed age-related changes for cognitive performance with improved reversal learning during the first three years due to practice followed by decline in the final year with the onset of old age. Cognitive decline was steeper and began earlier for females compared to males. Based on cognitive trajectories which were highly variable across individuals, we categorized marmosets as cognitively impaired or non-impaired. Mild amyloid β (Aβ) deposition was observed in the dlPFC and CA1 but did not differ between males and females. Aβ burden was higher in cognitively impaired compared to non-impaired marmosets, but this difference did not reach statistical significance. However, cognitively impaired marmosets showed signs of accelerated brain aging including higher astrogliosis associated with Aβ deposits and dendritic spine loss, both in the dlPFC and hippocampus. In addition, increased activation and changes in microglial phenotypes were observed in the dlPFC. Like humans, aging marmosets displayed intra-individual variability in cognitive decline, neurodegeneration and neuronal aging. Female marmosets showed greater vulnerability to cognitive impairment, which is translationally relevant to the higher prevalence of AD in women. While trajectories of cognitive decline did not strongly map onto Aβ burden for females or males, evidence of accelerated brain aging was found in the impaired marmosets. This study highlights the value of marmosets as a nonhuman primate model of AD to identify vulnerabilities to aging pathologies, such as sex differences.

EDITORIAL article Front. Endocrinol., 31 January 2023Sec. Neuroendocrine Science Volume 14 - 2023 | https://doi.org/10.3389/fendo.2023.1129203

The investigation of neurobiological and neuropathological changes that affect synaptic integrity and function with aging is key to understanding why the aging brain is vulnerable to Alzheimer's disease. We investigated the cellular characteristics in the cerebral cortex of behaviorally characterized marmosets, based on their trajectories of cognitive learning as they transitioned to old age. We found increased astrogliosis, increased phagocytic activity of microglial cells and differences in resting and reactive mi-croglial cell phenotypes in cognitively impaired compared to nonimpaired marmosets. Differences in amyloid beta deposition were not related to cognitive trajectory. However, we found age-related changes in density and morphology of dendritic spines in pyramidal neurons of layer 3 in the dorsolateral pre-frontal cortex and the CA1 field of the hippocampus between cohorts. Overall, our data suggest that an accelerated aging process, accompanied by neurodegeneration, that takes place in cognitively impaired aged marmosets and affects the plasticity of dendritic spines in cortical areas involved in cognition and points to mechanisms of neuronal vulnerability to aging. (c) 2022 Elsevier Inc. All rights reserved.

Olfactory dysfunction has been identified as an early biomarker for dementia risk but has rarely been assessed in nonhuman primate models of human aging. To better characterize common marmosets as such models, we assessed olfactory discrimination performance in a sample of 10 animals (5 females), aged 2.5-8.9 years old. The monkeys were proficient in the discrimination and reversal of visual stimuli but naïve to odor stimuli. For olfactory discrimination, the monkeys performed a series of six discriminations of increasing difficulty between two odor stimuli. We found no evidence for an age-related decline as both young and older individuals were able to perform the discriminations in roughly the same number of trials. In addition, the older monkeys had faster responses than the younger animals. However, we noted that when adjusted for age, the speed of acquisition of the first discrimination in the olfactory modality was inversely correlated to the speed of acquisition of their first discrimination of two visual stimuli months earlier. These results suggest that marmosets may compensate for sensory deficits in one modality with higher sensory performance in another. These data have broad implications for the assessment of age-related cognitive decline and the categorization of animals as impaired or nonimpaired.

Longitudinal studies are essential to understand healthy and pathological neurocognitive aging such as Alzheimer’s Disease, but longitudinal designs are rare in both humans and non-human primate models of aging because of the difficulty of tracking cognitive change in long-lived primates. Common marmosets ( Callithrix jacchus ) are uniquely suited for aging studies due to their naturally short lifespan (10- 12 years), sophisticated cognitive and social abilities and Alzheimer Disease-like neuropathology. We report the first longitudinal study of cognitive aging in marmosets (N=28) as they transitioned from middle- (~ 5 years) to old age (~ 9 years). We characterized aging trajectories using reversal learning with different stimuli each year. Marmosets initially improved on cognitive performance due to practice, but worsened in the final year, suggesting the onset of age-related decline. Cognitive impairment emerged earlier in females than males and was more prominent for discrimination than for reversal learning. Sex differences in cognitive aging could not be explained by differences in motivation or motor abilities, which improved or remained stable across aging. Likewise, males and females did not differ in aging trajectories of overall behavior or reactivity to a social stressor, with the exception of a progressive decline in the initiation of social behavior in females. Patterns of cognitive aging were highly variable across marmosets of both sexes, suggesting the potential for pathological aging for some individuals. Future work will link individual cognitive trajectories to neuropathology in order to better understand the relationships between neuropathologic burden and vulnerability to age-related cognitive decline in each sex.

Aromatase inhibitors (AIs) are a class of drugs commonly given to patients with estrogen receptor (ER)-dependent breast cancers to reduce estrogenic stimulation. However, AIs like Letrozole are associated with negative side effects such as cognitive deficits, sleep disturbances and hot flashes. We have previously shown that these negative effects can be recapitulated in common marmosets (Callithrix jacchus) treated with Letrozole (20 μg daily) for 4 weeks and that marmosets treated with Letrozole show increased levels of estradiol in the hippocampus (Gervais et al., 2019). In order to better understand the mechanisms through which AIs affect cognitive function and increase steroid levels in the hippocampus, we used bulk, paired-end RNA-sequencing to examine differentially expressed genes among Letrozole-treated (LET; n = 8) and vehicle-treated (VEH; n = 8) male and female animals. Gene ontology results show significant reduction across hundreds of categories, some of the most significant being inflammatory response, stress response, MHC Class II protein complex binding, T-cell activation, carbohydrate binding and signaling receptor binding in LET animals. GSEA results indicate that LET females, but not LET males, show enrichment for hormonal gene sets. Based on the transcriptional changes observed, we conclude that AIs may differentially affect the sexes in part due to processes mediated by the CYP-450 superfamily. Ongoing studies will further investigate the longitudinal effects of AIs on behavior and whether AIs increase the risk of stress-induced neurodegeneration.

While humans exhibit a significant degree of neuropathological changes associated with deficits in cognitive and memory functions during aging, non-human primates (NHP) present with more variable expressions of pathological alterations among individuals and species. As such, NHP with long life expectancy in captivity offer an opportunity to study brain senescence in the absence of the typical cellular pathology caused by age-related neurodegenerative illnesses commonly seen in humans. Age-related changes at neuronal population, single cell, and synaptic levels have been well documented in macaques and marmosets, while age-related and Alzheimer's disease-like neuropathology has been characterized in additional species including lemurs as well as great apes. We present a comparative overview of existing neuropathologic observations across the primate order, including classic age-related changes such as cell loss, amyloid deposition, amyloid angiopathy, and tau accumulation. We also review existing cellular and ultrastructural data on neuronal changes, such as dendritic attrition and spine alterations, synaptic loss and pathology, and axonal and myelin pathology, and discuss their repercussions on cellular and systems function and cognition.

Aging across the Primate Order is poorly understood because ages of individuals are often unknown, there is a dearth of aged animals available for study, and because aging is best characterized by longitudinal studies which are difficult to carry out in long‐lived species. The human population is aging rapidly, and advanced age is a primary risk factor for several chronic diseases and conditions that impact healthspan. As lifespan has increased, diseases and disorders of the central nervous system (CNS) have become more prevalent, and Alzheimer's disease and related dementias have become epidemic. Nonhuman primate (NHP) models are key to understanding the aging primate CNS. This Special Issue presents a review of current knowledge about NHP CNS aging across the Primate Order. Similarities and differences to human aging, and their implications for the validity of NHP models of aging are considered. Topics include aging‐related brain structure and function, neuropathologies, cognitive performance, social behavior and social network characteristics, and physical, sensory, and motor function. Challenges to primate CNS aging research are discussed. Together, this collection of articles demonstrates the value of studying aging in a breadth of NHP models to advance our understanding of human and nonhuman primate aging and healthspan.

Age-related cognitive decline has been extensively studied in humans, but the majority of research designs are cross-sectional and compare across younger and older adults. Longitudinal studies are necessary to capture variability in cognitive aging trajectories but are difficult to carry out in humans and long-lived nonhuman primates. Marmosets are an ideal primate model for neurocognitive aging as their naturally short lifespan facilitates longitudinal designs. In a longitudinal study of marmosets tested on reversal learning starting in middle-age, we found that, on average, the group of marmosets declined in cognitive performance around 8 years of age. However, we found highly variable patterns of cognitive aging trajectories across individuals. Preliminary analyses of brain tissues from this cohort also show highly variable degrees of neuropathology. Future work will tie together behavioral trajectories with brain pathology and provide a window into the factors that predict age-related cognitive decline.

Nonhuman primates (NHPs) are an essential research model for gaining a comprehensive understanding of the neural mechanisms of neurocognitive aging in our own species. In the present study, we used resting state functional connectivity (rsFC) to investigate the relationship between prefrontal cortical and striatal neural interactions, and cognitive flexibility, in unanaesthetized common marmosets (Callithrix jacchus) at two time points during late middle age (8 months apart, similar to a span of 5–6 years in humans). Based on our previous findings, we also determine the reproducibility of connectivity measures over the course of 8 months, particularly previously observed sex differences in rsFC. Male marmosets exhibited remarkably similar patterns of stronger functional connectivity relative to females and greater cognitive flexibility between the two imaging time points. Network analysis revealed that the consistent sex differences in connectivity and related cognitive associations were characterized by greater node strength and/or degree values in several prefrontal, premotor and temporal regions, as well as stronger intra PFC connectivity, in males compared to females. The current study supports the existence of robust sex differences in prefrontal and striatal resting state networks that may contribute to differences in cognitive function and offers insight on the neural systems that may be compromised in cognitive aging and age-related conditions such as mild cognitive impairment and Alzheimer’s disease.

Executive function (EF) is a complex construct that reflects multiple higher-order cognitive processes such as planning, updating, inhibiting and set-shifting. Decline in these functions is a hallmark of cognitive ageing in humans, and age differences and changes in EF correlate with age-related differences and changes in association cortices, particularly the prefrontal areas. Here, we review evidence for age-related decline in EF and associated neurobiological changes in prosimians, New World and Old World monkeys, apes and humans. While EF declines with age in all primate species studied, the relationship of this decline with age-related alterations in the prefrontal cortex remains unclear, owing to the scarcity of neurobiological studies focusing on the ageing brain in most primate species. In addition, the influence of sex, vascular and metabolic risk, and hormonal status has rarely been considered. We outline several methodological limitations and challenges with the goal of producing a comprehensive integration of cognitive and neurobiological data across species and elucidating how ageing shapes neurocognitive trajectories in primates with different life histories, lifespans and brain architectures. Such comparative investigations are critical for fostering translational research and understanding healthy and pathological ageing in our own species.This article is part of the theme issue ‘Evolution of the primate ageing process’.