BACKGROUND:Growing evidence suggests glucagon-like peptide-1 receptor agonists (GLP-1RAs) may represent a novel potential pharmacotherapeutic tool for alcohol use disorder (AUD). The objective of this study is to examine the association between GLP-1RA prescriptions and alcohol use. METHODS:This cohort study used a cross-sectional measure of alcohol consumption and longitudinal electronic health record (EHR) data collected between 1981 and October 2023 from NIH's All of Us Research Program, a large program to recruit and collect surveys, EHR, genomic, and wearable data from a wide array of Americans. Among 15,447 participants with at least two recorded GLP-1RA prescriptions on separate days, we created three groups based on the timing of Alcohol Use Disorders Identification Test-Consumption (AUDIT-C) responses relative to first GLP-1RA prescription. This resulted in 3650 with current GLP-1RA prescriptions, 5642 with future GLP-1RAs (primary comparison group), and 544 with previous GLP-1RAs. AUDIT-C scores were compared across these groups and to propensity-score matched comparison groups. RESULTS:Those with current GLP-1RA prescriptions had statistically significant but modestly lower AUDIT-C scores compared with those with future prescriptions (incidence rate ratio [IRR] = 0.95; 95% CI: 0.91-0.99; p = 0.01). Participants with a previous GLP-1RA prescription had lower AUDIT-C scores compared with those with future prescriptions, but this difference was not statistically significant. Results were similar using a matched comparison group with the current GLP-1RA group (IRR = 0.89; 95% CI: 0.85-0.93; p ≤ 0.001) and no significant difference for the previous prescription group. Analysis of individual AUDIT-C questions shows a significant association with GLP-1RA prescriptions and frequency of drinking but not drinks per occasion or binge drinking. CONCLUSIONS:This study's findings indicate that GLP-1RAs may reduce alcohol consumption by decreasing use frequency. Experimental studies and randomized controlled trials are needed to test the mechanisms and potential efficacy of GLP-1RAs in people with AUD.

Importance:Alcohol use is a leading cause of morbidity and mortality worldwide. Growing evidence suggests that glucagon-like peptide-1 receptor agonists (GLP-1RAs) may represent a novel potential pharmacotherapeutic tool for alcohol use disorder (AUD). Objective:To examine the association between GLP-1RA prescriptions and alcohol use. Design:This cohort study used a cross-sectional measure of alcohol consumption and longitudinal electronic health record (EHR) data collected between 1981 and October 2023 from NIH's All of Us Research Program participants. Setting:All of Us is a large program to recruit and collect surveys, EHR, genomic, and wearable data from a wide array of Americans. The data presented here are from the All of Us Curated Data Repository version 8. Participants:393,596 All of Us participants with EHR data recruited across the United States. Exposure:At least two GLP-1RA prescription records in the EHR. Main Outcomes and Measures:Alcohol Use Disorders Identification Test (AUDIT-C) scores and responses to individual AUDIT-C questions. Results:Among 15,447 participants with at least two recorded GLP-1RA prescriptions on separate days, 3650 had active GLP-1RA prescriptions, 5642 would have future GLP-1RA prescriptions (primary comparison group), and 544 had former GLP-1RA prescriptions. Those with active GLP-1RA prescriptions had statistically significant but modestly lower AUDIT-C scores on average compared with those with future prescriptions (incidence rate ratio [IRR] of 0.95; 95% CI, 0.91-0.99; P = 0.01). Participants with a former GLP-1RA prescription had lower AUDIT-C scores compared with those with future prescriptions, but this difference was not statistically significant. Results were similar using a propensity-score matched comparison group with a lower average AUDIT-C score for the current GLP-1RA group (IRR = 0.89; 95% CI, 0.85-0.93; P = <0.001) and no significant difference for the former prescription group. Analysis of individual AUDIT-C questions shows a significant association with GLP-1RA prescriptions and frequency of drinking but not drinks per occasion or binge drinking. Conclusions and Relevance:This study's findings indicate that GLP-1RAs may reduce alcohol consumption by decreasing use frequency. Experimental studies and randomized controlled trials are needed to test the mechanisms and potential efficacy of GLP-1RAs in people with AUD.

The choroid plexus (CP), a vital component in the brain’s ventricles, is crucial for cerebrospinal fluid (CSF) production and maintenance of the brain’s physiological environment. It plays a key role in regulating neuroinflammatory responses, clearing harmful substances, producing neurotrophic factors and signaling molecules, and forming blood-CSF barrier. Consequently, changes to the CP’s structural integrity could disrupt brain homeostasis and lead to cognitive impairment. Indeed, recent research has highlighted CP alterations in aging as well as in various cognitive disorders including Alzheimer’s and Parkinson’s diseases. This study investigates the correlation between the macrostructural and microstructural integrity of the CP and longitudinal cognitive changes in cognitively unimpaired individuals. 116 cognitively unimpaired participants from the BLSA and GESTALT underwent advanced MRI, including relaxometry (T1 and T2) and diffusion tensor imaging (DTI), enabling the quantification of CP volume, fractional anisotropy (FA), and mean diffusivity (MD). Participants received cognitive assessments at visits preceding and concurrent with MRI scans (320 visits total) to measure memory, attention, executive function, verbal fluency, and processing speed (Table 1). Linear mixed-effects models were used to analyze the association between MRI metrics and longitudinal cognitive changes, adjusting for age, sex, education, and race. Our study found that larger CP volume, elevated T1 and T2 values, and higher MD values, all indicative of compromised macrostructural or microstructural integrity, were associated with faster declines in various cognitive domains (Table 2). Particularly, the association was found in memory, verbal fluency, and processing speed for CP volume; in attention, executive function, and processing speed for T1 and T2 (Figure 1); and in attention, executive function, and processing speed for MD. Conversely, lower FA values, representing deteriorated microstructural integrity, correlated with accelerated decline in memory, attention, verbal fluency, and processing speed. This study highlights a significant association between CP characteristics and cognitive decline in cognitively unimpaired individuals. These findings underscore the importance of the CP in cognitive health and emphasize the need for further research into the role of the CP in healthy aging, potentially opening new pathways for early detection and intervention in cognitive decline.

With global dementia rates rising sharply, there is an urgent need for sensitive biomarkers to detect cognitive changes and predict dementia risk. White matter degeneration, especially axonal loss, is increasingly recognized as an early hallmark of Alzheimer's disease (AD), but its temporal trajectory and its relationship with cognition have not been established. We utilized a novel MRI-derived Axonal Density Index (ADI) to longitudinally investigate axonal degeneration and cognitive decline in the ADNI cohort. Linear mixed-effects models showed cognitively impaired individuals had lower baseline ADI and faster axonal degeneration compared to cognitively normal subjects. In cognitively impaired individuals, higher baseline ADI predicted slower prospective cognitive deterioration and lower dementia risk, while greater longitudinal ADI declines correlated with cognitive worsening and increased dementia risk. Notably, ADI outperformed cerebrospinal fluid biomarkers of AD pathology in predicting cognitive outcomes. Our original findings position axonal degeneration as an early AD feature and ADI as a promising biomarker for early detection, disease phenotyping and monitoring, and intervention targets.

Insulitis, a hallmark of inflammation preceding autoimmune type 1 diabetes, leads to the eventual loss of functional beta cells. However, functional beta cells can persist even in the face of continuous insulitis. Despite advances in immunosuppressive treatments, maintaining functional beta cells to prevent insulitis progression and hyperglycaemia remains a challenge. The cannabinoid type 1 receptor (CB1R), present in immune cells and beta cells, regulates inflammation and beta cell function. Here, we pioneer an ex vivo model mirroring human insulitis to investigate the role of CB1R in this process. CD4+ T lymphocytes were isolated from peripheral blood mononuclear cells (PBMCs) from male and female individuals at the onset of type 1 diabetes and from non-diabetic individuals, RNA was extracted and mRNA expression was analysed by real-time PCR. Single beta cell expression from donors with type 1 diabetes was obtained from data mining. Patient-derived human islets from male and female cadaveric donors were 3D-cultured in solubilised extracellular matrix gel in co-culture with the same donor PBMCs, and incubated with cytokines (IL-1β, TNF-α, IFN-γ) for 24–48 h in the presence of vehicle or increasing concentrations of the CB1R blocker JD-5037. Expression of CNR1 (encoding for CB1R) was ablated using CRISPR/Cas9 technology. Viability, intracellular stress and signalling were assayed by live-cell probing and real-time PCR. The islet function measured as glucose-stimulated insulin secretion was determined in a perifusion system. Infiltration of immune cells into the islets was monitored by microscopy. Non-obese diabetic mice aged 7 weeks were treated for 1 week with JD-5037, then euthanised. Profiling of immune cells infiltrated in the islets was performed by flow cytometry. CNR1 expression was upregulated in circulating CD4+ T cells from individuals at type 1 diabetes onset (6.9-fold higher vs healthy individuals) and in sorted islet beta cells from donors with type 1 diabetes (3.6-fold higher vs healthy counterparts). The peripherally restricted CB1R inverse agonist JD-5037 arrested the initiation of insulitis in humans and mice. Mechanistically, CB1R blockade prevented islet NO production and ameliorated the ATF6 arm of the unfolded protein response. Consequently, cyto/chemokine expression decreased in human islets, leading to sustained islet cell viability and function. These results suggest that CB1R could be an interesting target for type 1 diabetes while highlighting the regulatory mechanisms of insulitis. Moreover, these findings may apply to type 2 diabetes where islet inflammation is also a pathophysiological factor. Transcriptomic analysis of sorted human beta cells are from Gene Expression Omnibus database, accession no. GSE121863, available at https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM3448161 .

Senescent cells accumulate with aging and are associated with several age-associated diseases and functional declines. Eliminating senescent cells with senolytics improves aging phenotypes in mouse models and may improve the health of people with chronic diseases. To date, very few senotherapeutic (senolytics and senomorphics) compounds have been identified. In a recent study, we reported that gingerenone A (GinA) has a senolytic effect via mechanisms including the activation of caspase-3 activity and apoptotic cell death. In this study, we investigated whether GinA has senotherapeutic properties in a mouse model of senescence. Moreover, we modified GinA with eicosapentaenoic acid (EPA) esters (GinA-EPA) or docosahexaenoic acid (DHA) esters (GinA-DHA) to generate modified gingerenone A (modGinA) that could enhance GinA effects. We found that both GinA and modGinA induced biochemical and histological changes consistent with anti-inflammatory, senolytic, and senomorphic effects, leading to improved metabolic and mitochondrial functions.

Identifying a reliable biomarker for amyotrophic lateral sclerosis (ALS) is crucial for clinical practice. Here, in this cross-sectional study, we used the Olink Explore 3072 platform to investigate plasma proteomics as a biomarker tool for this neurodegenerative condition. Thirty-three proteins were differentially abundant in the plasma of patients with ALS (n = 183) versus controls (n = 309). We replicated our findings in an independent cohort (n = 48 patients with ALS and n = 75 controls). We then applied machine learning to create a model that diagnosed ALS with high accuracy (area under the curve, 98.3%). By analyzing plasma samples from individuals before ALS symptoms emerged, we estimated the age of clinical onset and showed that the disease process-impacting skeletal muscle, nerves and energy metabolism-occurs years before symptoms appear. Our research suggests that plasma proteins can be a biomarker for this fatal disease and offers molecular insights into its prodromal phase.

The brainstem plays a vital role in regulating blood pressure, and disruptions to its neural pathways have been linked to hypertension. However, it remains unclear whether subtle microstructural changes in the brainstem are associated with an individual’s blood pressure status. This exploratory, cross-sectional study investigated the relationship between brainstem microstructure, myelination, and hypertensive status in 116 cognitively unimpaired adults (aged 22–94 years). Advanced MRI techniques, including relaxometry (R1, R2) and myelin water fraction (MWF) analysis, were employed to assess microstructural integrity and myelin content in ten brainstem subregions. Our results revealed significant associations between higher microstructural damage or lower myelin content (indicated by lower R1, R2, or MWF values) and hypertensive status, particularly in the midbrain tegmentum. Notably, combining these MRI metrics yielded high classification accuracy (AUC > 0.85). Our findings suggest a potential link between disrupted brainstem tissue integrity, myelin content, and elevated blood pressure, warranting further longitudinal investigations to explore this relationship.

The Locus Coeruleus (LC) is a critical brain region affected by neurodegenerative diseases and aging. Despite its importance, in-vivo investigations of age-related LC degeneration and association with cognitive decline have been limited. We employed magnetic resonance relaxometry, namely the Bayesian Monte-Carlo analysis of multicomponent driven equilibrium single pulse observation of T1 and T2 (BMC-mcDESPOT) MRI method, to estimate microstructural integrity represented by longitudinal (R1) and transverse (R2) relaxation rates, as well as Myelin Water Fraction (MWF) in the LC of a diverse cohort of 120 cognitively unimpaired individuals aged 22 to 94 years. BMC-mcDESPOT offers high spatial resolution and is effective for mapping detailed microstructural changes within the LC. We examined age-related differences in LC microstructure, their associations with cognitive changes, and the spatial variation of these microstructural changes within the LC, exploring their distinctive contributions to cognitive decline. LC-R2 values declined significantly with age, particularly in the rostral-middle regions. LC-R1 and LC-MWF values showed significant positive correlations with cross-sectional memory scores. Longitudinally, the rostra-middle LC-R2 values showed an age-moderated effect, with lower values predicting steeper memory decline at advanced ages. Quantitative MR relaxometry reveals that LC microstructural integrity declines with age and is predictive of cognitive decline, particularly in memory. Our MR relaxometry biomarkers, especially in the rostral LC, serve as sensitive imaging biomarkers of early structural alterations and cognitive declines in aging.

IntroductionTotal body metabolism continuously adapts to match energy supply with demand. During exercise metabolic alterations occur because skeletal muscles require a continuous supply of newly generated ATP to match the demand of the intensity of the exercise, and products of muscle metabolism must be eliminated. The metabolic and energetic flexibility greatly impact maximum physical fitness and exercise duration, as well as the speed of elimination of metabolism end-products. However, so far, the temporal profiling of metabolomic changes in response to exercise of persons with different fitness levels remains relatively unexplored. This study examined metabolic changes during each person’s peak aerobic exercise and one-hour post-exercise recovery in 29 Baltimore Longitudinal Study of Aging (BLSA) participants.MethodsBlood samples were collected at baseline, and at 3-min intervals during both incremental exercise on a treadmill until exhaustion and during recovery. Participants were classified based on the stage when they reached exhaustion as low fitness (LF, completing up to 3 treadmill incremental stages) or high fitness (HF, completing up to 7 incremental stages). The time course of exercise-associated changes in the circulating metabolome were mapped and unique metabolomic trajectories were identified with likelihood-ratio testing and hierarchical clustering.ResultsThe HF group had rapid clearance of bile and amino acids at exercise onset, along with effective clearance of triacylglycerols and glycerophospholipids during recovery. In contrast, the LF group had much reduced clearance of these metabolites and had persistent elevation of triacylglycerols and glycerophospholipids.DiscussionThese findings highlight differences in bile acid clearance and purine metabolism in people of differing fitness levels and provide novel insights into the role of metabolic adaptive responses to aerobic exercise assessed through circulating metabolomic measures.

Insulin Resistance (IR) is implicated in brain aging and Alzheimer’s disease (AD) pathogenesis. Dietary changes may promote brain health in older adults with metabolic abnormalities. An extensive animal literature suggests pro-cognitive and beneficial systemic and brain effects of intermittent fasting (IF) that may mitigate AD risk. We conducted a randomized clinical trial comparing brain effects and the potential for AD biomarker modulation of IF and a continuous diet. Forty overweight, cognitively intact individuals > 55 years old with peripheral IR were randomized (1:1) to 5:2 IF (2 days 480 Kcal/day; 5 non-restricted days) or Healthy Living (HL) diet by USDA recommendations for 8 weeks. Both IF and HL improved biomarkers of brain IR in neuron-derived extracellular vesicles (NDEVs), decreased regional BrainAGE (brain-age-gap estimate on structural MRI) in the anterior cingulate and ventromedial prefrontal cortex, reduced glucose concentration on brain magnetic resonance spectroscopy (MRS) indicating optimized metabolism, and improved executive function and memory. Moreover, both diets decreased weight, BMI, and waist circumference suggesting high compliance, and HOMA2-IR indicating IR alleviation. They also increased blood beta-hydroxybutyrate and acetoacetate suggesting increased ketogenesis. Although effects of IF and HL were statistically comparable, greater numerical improvements were observed with IF for several measures of brain health, cognition, and peripheral metabolism. However, cerebrospinal fluid (CSF) and NDEVs showed no changes in AD biomarkers (Aβ 42 , total Tau, p181-Tau, GFAP). In exploratory analysis, effects were moderated by polymorphisms in ApoE ε4 genotype, with cognitive benefits and decreased MRS glucose solely in ε4 non-carriers; ε4 carriers showed no cognitive benefits, no decreases in MRS glucose, but increased CSF Aβ 42 /Aβ 40 ratio, NfL and GFAP. Beneficial effects of HL and, especially, IF on NDEV-derived brain IR, BrainAGE, brain glucose concentration, executive function and memory indicate that healthy diets may improve brain health even over short periods of time, albeit with no evidence for AD cascade modulation. Genetic factors, such as ApoE ε4 polymorphisms, may influence the brain’s response to diets. Future clinical trials encompassing IF and other dietary interventions and stratified by ApoE ε4 carrier status may provide avenues for slowing the pace of brain aging.

ABSTRACT The study of biomarkers in biofluids and tissues expanded our understanding of the biological processes that drive physiological and functional manifestations of aging. However, most of these studies were limited to examining one biological compartment, an approach that fails to recognize that aging pervasively affects the whole body. The simultaneous modeling of hundreds of metabolites and proteins across multiple compartments may provide a more detailed picture of healthy aging and point to differences between chronological and biological aging. Herein, we report proteomic analyses of plasma and urine collected in healthy men and women, age 22–92 years. Using these data, we developed a series of metabolomic and proteomic predictors of chronological age for plasma, urine, and skeletal muscle. We then defined a biological aging score, which measures the departure between an individual's predicted age and the expected predicted age for that individual based on the full cohort. We show that these predictors are significantly and independently related to clinical phenotypes important for aging, such as inflammation, iron deficiency anemia, muscle mass, and renal and hepatic functions. Despite a different set of selected biomarkers in each compartment, the different scores reflect a similar degree of deviation from healthy aging in single individuals, thus allowing identification of subjects with significant accelerated or decelerated biological aging.

BACKGROUND:Cardiolipins (CL) are a mitochondria-specific family of phospholipids that play central roles in mitochondrial function. Imbalance in CL metabolism, especially excessive CL oxidation, leads to mitochondrial dysfunction, apoptosis, and inflammation, contributing to age-related diseases. As of yet no comprehensive methods have been developed to assess CL, oxidized CL (oxCL), and monolyso-CL (MLCL) species. RESULTS:To fill this critical research gap, we combined untargeted and targeted lipidomic approaches to analyze CL species in human skeletal muscle samples. The method enabled in-depth structural characterization using exact mass measurement followed by multistage fragmentation (MSn) to achieve unequivocal structural elucidation at the molecular species level and sn-position level for some species. This novel methodology identified intact mono- and di-oxygenated L4CL species and allowed the differentiation of isomeric 9/13-HODE-L3CL and 9(10)/12(13)-EpOME-L3CL with unprocessed total lipid extracts. Overall, 220 molecular species (125 CL, 30 oxCL, and 65 MLCL) were detected. Our method includes a quantitation strategy that leveraged on establishing three-leveled matrix-matched calibration curves normalized by using internal standard M4CL and its isotopologues M+1 (13C-M4CL) and M+3 (13C3-M4CL) respectively. The analytical performance was also evaluated and found to be highly sensitive with LLOQs at fmol levels and reproducible with precision RSD <20 % for the majority. SIGNIFICANCE AND NOVELTY:This is the first reported method to simultaneously provide broad coverage of CL, oxCL, and MLCL species from a single injection of total lipid extract in a complex biological sample. This method is also the first to demonstrate the presence of sn-positional isomers of CL in human skeletal muscle. It has been successfully applied to a pilot study of skeletal muscle biopsies and provided meaningful results. We anticipate the methodology will facilitate investigations of the cardiolipin lipidome leading to a better understanding of the complex and highly interactive biological processes that regulate mitochondria function and expand minor cardiolipin targets including oxCL and MLCL for biomarker discovery.

Aged pancreatic fibroblasts secrete GDF-15 and activate AKT signaling to promote pancreatic cancer growth, highlighting the critical role of aging-mediated changes in the pancreatic cancer microenvironment in driving tumor progression. Pancreatic cancer is more prevalent in older individuals and often carries a poorer prognosis for them. The relationship between the microenvironment and pancreatic cancer is multifactorial, and age-related changes in nonmalignant cells in the tumor microenvironment may play a key role in promoting cancer aggressiveness. Because fibroblasts have profound impacts on pancreatic cancer progression, we investigated whether age-related changes in pancreatic fibroblasts influence cancer growth and metastasis. Proteomics analysis revealed that aged fibroblasts secrete different factors than young fibroblasts, including increased growth/differentiation factor 15 (GDF-15). Treating young mice with GDF-15 enhanced tumor growth, whereas aged GDF-15 knockout mice showed reduced tumor growth. GDF-15 activated AKT, rendering tumors sensitive to AKT inhibition in an aged but not young microenvironment. These data provide evidence for how aging alters pancreatic fibroblasts and promotes tumor progression, providing potential therapeutic targets and avenues for studying pancreatic cancer while accounting for the effects of aging.Significance: Aged pancreatic fibroblasts secrete GDF-15 and activate AKT signaling to promote pancreatic cancer growth, highlighting the critical role of aging-mediated changes in the pancreatic cancer microenvironment in driving tumor progression. See related commentary by Isaacson et al., p. 1185Significance: Aged pancreatic fibroblasts secrete GDF-15 and activate AKT signaling to promote pancreatic cancer growth, highlighting the critical role of aging-mediated changes in the pancreatic cancer microenvironment in driving tumor progression. See related commentary by Isaacson et al., p. 1185

Context: Body composition and glucose metabolism change with aging. Whether different levels of body-mass-index (BMI) are needed to define diabetes risk across the adult lifespan is unknown. Objective: This work aimed to investigate whether BMI similarly reflects relative fat mass (FM) and diabetes risk across age groups. Methods: Participants without diabetes from the Baltimore Longitudinal Study of Aging (973 men, 1073 women), stratified by age (<50, 50-59, 60-69, >= 70 years) and categorized by either World Health Organization (WHO)-defined BMI categories (for normal weight, overweight or obesity) or BMI quartiles. The primary exposure was BMI. The primary outcome was diabetes incidence. The relationship of BMI to dual-energy x-ray absorptiometry-derived FM was also investigated in older vs younger participants. Results: The median (range) follow-up time was 7.1 years (range, 0-29.0 years). Within WHO-defined BMI categories, different age groups demonstrated significantly different FM percentage, FM/lean mass, and waist circumference (P < .05). WHO-defined BMI categories for overweight and obesity were generally related to higher diabetes risk compared to normal weight in all ages except 50 to 59 years. When BMI was categorized by quartiles, diabetes incidence increased dramatically beginning in quartile 2 (23-25 kg/m(2)) in older groups. BMI cutoffs with equivalent diabetes incidence rate as BMI 25 kg/m(2) and 30.0 kg/m(2) in individuals younger than 50 years were 22.7 kg/m(2) and 25.2 kg/m(2) for ages 50 to 59 years; 22.8 kg/m(2) and 25.0 kg/m(2) for ages 60 to 69 years; and 23.2 kg/m(2) and 25.8 kg/m(2) for ages 70 years and older, respectively. Conclusion: WHO-defined BMI categories do not reflect similar diabetes risk across the lifespan. Diabetes incidence is greater at lower levels of BMI in older adults and may lead to underestimation of diabetes risk with aging, particularly among those traditionally classified as normal-weight individuals.