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Background: Cardiovascular disease may be the main reason for stagnant growth in life expectancy in the United States since 2010. The American Heart Association recently released an updated algorithm for evaluating cardiovascular health (CVH)—Life’s Essential 8 (LE8) score. We aimed to quantify the associations of CVH levels, estimated by the LE8 score, with life expectancy in a nationally representative sample of US adults. Methods: We included 23 003 nonpregnant, noninstitutionalized participants aged 20 to 79 years who participated in the National Health and Nutrition Examination Survey from 2005 to 2018 and whose mortality was identified through linkage to the National Death Index through December 31, 2019. The overall CVH was evaluated by the LE8 score (range, 0–100), as well as the score for each component of diet, physical activity, tobacco/nicotine exposure, sleep duration, body mass index, non–high-density lipoprotein cholesterol, blood glucose, and blood pressure. Life table method was used to estimate life expectancy by levels of the CVH. Results: During a median of 7.8 years of follow-up, 1359 total deaths occurred. The estimated life expectancy at age 50 years was 27.3 years (95% CI, 26.1–28.4), 32.9 years (95% CI, 32.3–33.4), and 36.2 years (95% CI, 34.2–38.2) in participants with low (LE8 score Conclusions: Adhering to a high CVH, defined as the LE8 score, is related to a considerably increased life expectancy in US adults, but more research needs to be done in other races and ethnicities (eg, Hispanic and Asian).

Growing evidence has linked gut microbiota with regulation of adiposity. We aimed to examine whether the genetically determined relative abundance of gut microbial taxa was associated with long-term changes in adiposity and body composition among individuals who were overweight or obese in weight-loss diet interventions.The study included 692 participants with overweight or obese from the POUNDS Lost trial. We created a genetic risk score (GRS) for the relevant abundance of gut microbial taxa using 20 single nucleotide polymorphisms identified from a recent genome-wide association study. Body composition was assessed using dual-energy X-ray absorptiometry.Higher GRS for the relative abundance of gut microbial taxa was significantly associated with greater reductions in waist circumference, total fat mass (FM), whole-body total percentage of fat mass (FM%), and percentage of trunk fat (TF%) at 2 years (p = 0.022, 0.034, 0.023, 0.023, respectively). In addition, dietary protein significantly modified the association between GRS for gut microbial abundance and changes in total FM, FM%, and TF% (p-interactions = 0.04, 0.013, and 0.006, respectively) at 6-month, when the maximum weight loss was achieved, even though such interactions were attenuated at 2 years. In the average-protein diet group, a higher microbial abundance GRS was associated with greater reductions in total FM (p = 0.007), FM% (p = 0.002), and TF% (p 0.001) at 6 months, while no associations were found in the high-protein diet group (p 0.05).Our results suggest that the higher genetically determined relative abundance of gut microbial taxa may be related to long-term improvement of whole-body and central fatness and body composition in response to low-calorie diet interventions.

BACKGROUND: MicroRNAs (miRNAs) are short noncoding RNAs and important posttranscriptional regulators of gene expression. Adipose tissue is a major source of circulating miRNAs; adipose-related circulating miRNAs may regulate body fat distribution and glucose metabolism. OBJECTIVES: We investigated how changes in adipose-related circulating microRNAs-99/100 (miR-99/100) in response to lifestyle interventions were associated with improved body fat distribution and reductions of diabetogenic ectopic fat depots among adults with abdominal obesity. METHODS: This study included adults with abdominal obesity from an 18-mo diet and physical activity intervention trial. Circulating miR-99a-5p, miR-99b-5p, and miR-100-5p were measured at baseline and 18 mo; changes in these miRNAs in response to the interventions were evaluated. The primary outcomes were changes in abdominal adipose tissue [visceral (VAT), deep subcutaneous (DSAT), and superficial subcutaneous (SSAT) adipose tissue; cm(2)] (n = 144). The secondary outcomes were changes in ectopic fat accumulation in the liver (n = 141) and pancreas (n = 143). RESULTS: Greater decreases in miR-100-5p were associated with more reductions of VAT (β ± SE per 1-SD decrease: −9.63 ± 3.13 cm(2); P = 0.0025), DSAT (β ± SE: −5.48 ± 2.36 cm(2); P = 0.0218), SSAT (β ± SE: −4.64 ± 1.68 cm(2); P = 0.0067), and intrahepatic fat percentage (β ± SE: −1.54% ± 0.49%; P = 0.0023) after the interventions. Similarly, participants with greater decrease in miR-99a-5p had larger 18-mo reductions of VAT (β ± SE: −10.12 ± 3.31 cm(2) per 1-SD decrease; P = 0.0027) and intrahepatic fat percentage (β ± SE: −1.28% ± 0.52%; P = 0.015). Further, decreases in circulating miR-99b-5p (β ± SE: per 1-SD decrease: −0.44% ± 0.21%; P = 0.038) and miR-100-5p (β ± SE: −0.50% ± 0.23%; P = 0.033) were associated with a decrease in pancreatic fat percentage, as well as improved glucose metabolism and insulin secretion at 18 mo. CONCLUSIONS: Decreases in circulating miR-99-5p/100-5p expression induced by lifestyle interventions were related to improved body fat distribution and ectopic fat accumulation. Our study suggests that changes in circulating adipose-related miR-99-5p/100-5p may be linked to reducing diabetogenic fat depots in patients with abdominal obesity. This trial was registered at clinicaltrials.gov as NCT01530724.

AIMS: To examine whether changes in objectively measured physical activity (PA) are associated with weight loss and changes in body composition and fat distribution in response to weight-loss diet interventions. METHODS: This study included 535 participants with overweight/ obesity randomly assigned to 4 weight-loss diets varying in macronutrients. PA was measured objectively with pedometers, and body composition and fat distribution were measured using dual-energy X-ray absorptiometry (DEXA) scans and computed tomography (CT) scans at baseline, 6 months, and 24 months. RESULTS: From baseline to 6 months, when the maximum weight loss was achieved, each 1000 daily steps increment in PA was associated with a greater reduction in body weight (β[SE]=−0.48[0.11]) and waist circumference (β[SE]=−0.49[0.12]). Similar inverse associations were found in changes in body composition and fat distribution (P

Background Thioredoxin Interacting Protein (TXNIP) functions as a master regulator for glucose homeostasis. Hypomethylation at the 5’-cytosine-phosphate-guanine-3’ (CpG) site cg19693031 of TXNIP has been consistently related to islet dysfunction, hyperglycemia, and type 2 diabetes. DNA methylation (DNAm) may reveal the missing mechanistic link between obesity and type 2 diabetes. We hypothesize that baseline DNAm level at TXNIP in blood may be associated with glycemic traits and their changes in response to weight-loss diet interventions. Methods We included 639 adult participants with overweight or obesity, who participated in a 2-year randomized weight-loss diet intervention. Baseline blood DNAm levels were profiled by high-resolution methylC-capture sequencing. We defined the regional DNAm level of TXNIP as the average methylation level over CpGs within 500 bp of cg19693031. Generalized linear regression models were used for main analyses. Results We found that higher regional DNAm at TXNIP was significantly correlated with lower fasting glucose, HbA1c, and Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) at baseline (P 0.05). The interaction was attenuated to be non-significant at 2 years, presumably related to decreasing adherence to the diet intervention. Conclusions Our data indicate that higher regional DNAm level at TXNIP was significantly associated with better fasting glucose, HbA1c, and HOMA-IR; and people with higher regional DNAm levels benefited more in insulin and HOMA-IR improvement by taking the average-protein weight-loss diet.

Introduction: A previous epigenome-wide association study has causally linked DNA methylation (DNAm) at the SREBF1 gene (cg11024682) with obesity and lipids. However, little is known about whether DNAm at SREBF1 is associated with long-term changes in body adiposity and composition. Hypothesis: We hypothesized that participants with different DNAm at SRBF1 might respond differently to dietary weight-loss interventions on changes in body adiposity and composition. Methods: The current study included 314 individuals with overweight or obese, who participated in POUNDS Lost: a 2-year randomized dietary weight-loss trial. The blood DNAm level was profiled by methylC-capture sequencing at baseline. Regional DNAm at SREBF1 was calculated as the average methylation level over CpGs within ±250 bp of cg11024682. Body composition, including total fat mass (FM), percentage of FM (FM%), total fat-free mass (FFM), percentage of FFM (FFM%), and percentage of trunk fat (TF%) were measured by dual-energy X-ray absorptiometry (DEXA) at baseline, 6 months, and 2 years. Results: Lower regional DNAm at SREBF1 was significantly associated with changes in body composition across 2 years ( Table ). At 6 months, per SD lower regional DNAm at SREBF1 was significantly associated with greater reductions in FM (β [SE] -0.23 [0.07], p=0.002), FM% (-0.30 [0.09], p Conclusions: Overweight and obese individuals with a lower regional DNAm at SREBF1 achieved greater improvement in body composition across the 2-year intervention, independent of concurrent weight loss, suggesting DNAm at SREBF1 is predictive of individuals’ response to treatment.

Introduction: The microRNA-122 (miR-122) is widely involved in the post-transcriptional regulation of glucose and lipid metabolism, and cardiovascular system. Hypothesis: We assessed the hypothesis that decrease in circulating miR-122-5p would be associated with the improvement in cardiovascular health (CVH) score and each component health metric in response to dietary weight-loss interventions. Methods: Circulating miR-122-5p levels were measured at baseline and 6 months among 509 participants with overweight or obesity from the POUNDS LOST trial. We evaluated the CVH score using the newly updated algorithm -life’s essential 8 and calculated the composite score from the available biological health factors (body mass index [BMI], blood lipids, blood glucose, and blood pressure) across the 2-year intervention period. Results: At baseline, lower levels of miR-122-5p were associated with higher CVH scores (PP time*miR-122-5p =0.001). We also found a significant interaction between change in miR-122-5p and dietary fat intake on the 6-months change in CVH ( P = 0.025); a greater decrease in miR-122-5p showed a greater improvement in CVH at 6 months among participants assigned to a low-fat diet. Conclusions: In conclusion, participants with decreased circulating miR-122-5p levels in response to dietary weight-loss interventions had significant improvement in CVH and achieved CVH benefits most strongly when consuming a low-fat diet.

Background: Cardiovascular disease (CVD) may be the main reason for stagnant growth in life expectancy in the US since 2010. The American Heart Association (AHA) recently released updated algorithm for evaluating cardiovascular health (CVH)--life’s essential 8 (LE8) score. Hypothesis: We aimed to quantify the associations of CVH levels, estimated by the LE8 score, with life expectancy in a nationally representative sample of US adults. Methods: We included 23003 adults who participated in the National Health and Nutrition Examination Survey (NHANES) 2005-2018 and whose mortality was identified through linkage to the National Death Index through December 31, 2019. Life table method was used to estimate life expectancy by levels of the CVH. The cause-specific decomposition of the life expectancy differences was evaluated by the Arriaga’s method. Results: During a median of 7.8 years of follow-up, 1359 total deaths occurred (772 in men and 587 in women). The estimated life expectancy at age 50 was 25.5 years (95% CI, 23.8-27.2), 31.2 years (95% CI, 30.4-31.9) and 33.1 years (95% CI, 29.8-36.2) in men with poor, intermediate and ideal CVH, respectively. The corresponding estimated life expectancy at age 50 in women was 29.5 years (95% CI, 27.8-31.1), 34.2 years (95% CI, 33.5-34.9) and 38.4 years (95% CI, 35.4-41.0), respectively. Equivalently, men and women with ideal CVH had an average 7.5 (95% CI, 3.5-11.5) and 8.9 (95% CI, 5.2-12.5) more years of life expectancy at age 50, respectively, compared with their counterparts with poor CVH. In men, on average, 41.8 % of the gained life expectancy at age 50 from adhering to ideal CVH was attributable to reduced CVD death. The corresponding percentage was 44.1 % in women. Similarly significant associations of CVH with life expectancy were observed in Non-Hispanic Whites and Non-Hispanic Blacks but not in Mexican Americans. Conclusions: Adhering to an ideal CVH, defined as the LE8 score, is related to a considerably increased life expectancy in the US adults.

Introduction: Epigenome-wide association studies identified that DNA methylation (DNAm) at the ABCG1 gene (cg06500161) was associated with obesity. However, little is known about whether DNAm at ABCG1 is associated with long-term changes in adiposity and body fat distribution in response to dietary weight-loss interventions. Hypothesis: We hypothesized that participants with varying DNAm at ABCG1 might respond differently to dietary weight-loss interventions on long-term changes in adiposity and body fat distribution. Methods: The current study included 673 participants with overweight/obesity, who were randomly assigned to 1 of 4 diets varying in macronutrient components. Blood DNAm level was profiled by a high-resolution methylC-capture sequencing at baseline. The regional DNAm at ABCG1 was defined as the average methylation level over CpGs within ± 250 bp of cg06500161. Two-year changes in adiposity and Computed Tomography (CT)-measured body fat distribution were calculated. Results: We found that regional DNAm at ABCG1 significantly interacted with dietary protein intake on long-term changes in body weight and waist circumference (WC) at 6 months and 2 years (P-interactionFigure ). Among participants assigned to an average-protein (15%) diet, lower regional DNAm at ABCG1 was associated with greater reductions in body weight and WC at 6 months and 2 years, while opposite associations were found among those assigned to a high-protein (25%) diet. Similar interactions were also observed for body fat distribution, including visceral adipose tissue, subcutaneous adipose tissue, deep subcutaneous adipose tissue, and total adipose tissue at 6 months and 2 years (P-interaction Conclusion: Our data indicate that DNAm at ABCG1 interacted with dietary protein intake on long-term changes in adiposity and body fat distribution. Participants with hypomethylation at ABCG1 benefited more in long-term improvement in adiposity and body fat distribution by consuming an average-protein diet.

Introduction: MicroRNA-19 (MiR-19) plays a critical role in cardiac development and cardiovascular disease (CVD). Hypothesis: We assessed the hypothesis that change in circulating miR-19 would be associated with changes in 10-year atherosclerotic CVD (ASCVD) risk and individual cardiovascular risk factors in a dietary weight-loss intervention. Methods: This study included 509 participants with overweight or obesity from the 2-year weight-loss diet intervention study (the POUNDS LOST trial) and with available data on circulating miR-19a/b-3p at baseline and 6 months. The primary outcome was the 6-month change in the ASCVD risk score, which estimates the 10-year probability of hard ASCVD events. Secondary outcomes were the change in each ASCVD component including blood pressure, lipid profiles, insulin resistance, glucose metabolism, and adiposity measures. Change in habitual physical activity and sleep disturbance were assessed from baseline to 6 months. Results: Circulating miR-19a/b-3p levels significantly decreased during the initial 6-month dietary intervention period (P=0.008, 0.0004, respectively). We found that greater decreases in log-transformed miR-19a/b-3p were related to a greater reduction in ASCVD risk (β[SE]=0.35 [0.13], P=0.01 for miR-19a-3p; β[SE]=0.33 [0.13], P=0.01 for miR-19b-3p) over 6 months. Similar positive associations were observed for changes in diastolic blood pressure with miR-19a/b-3p. Moreover, we found that sleep disturbance was significantly associated with reductions in miR-19a/b-3p from baseline to 6 months (P trend Conclusions: In conclusion, decreased circulating miR-19 levels induced by dietary weight-loss interventions were related to a significant reduction in ASCVD risk, and such associations were more evident in people with no/slight amount of sleep disturbance and an increase in physical activity.

Introduction: The American Heart Association recently introduced an enhanced approach to assess cardiovascular health (CVH): Life’s essential 8 (LE8). Sleep was included as a new component. The current study sought to analyze the relationship between levels of CVH, indicated by the LE8 score, and risks of coronary heart disease (CHD), stroke, and cardiovascular disease (CVD). Hypothesis: We hypothesized that higher CVH, indicated by the LE8 score, was associated with lower risks of CHD, stroke, and CVD; LE8 might also interact with genetic susceptibility. Methods: The current study included 137,794 participants free of CVD from the UK Biobank. CVH was scored at baseline by LE8 (DASH diet score, physical activity, nicotine exposure, sleep health, body mass index, blood lipids, blood glucose, and blood pressure). The LE8 score ranged from 0 to 100; and was categorized as low (0-49), moderate (50-79), and high (80-100). Weighted genetic risk scores (GRS) for CHD and stroke were calculated. Cox proportional hazard model was used to assess the relationship between LE8 and incident CVD outcomes, and potential interaction with age, gender, and GRS. We also compared the performance of LE8 with Life’s Simple 7 (LS7) using Harrell’s C-index. Results: During a median of 10 years, 8,595 CVD cases (6,968 CHD and 1,948 strokes) were documented. A higher LE8 score was associated with remarkably lower risks of CHD, stroke, and CVD (p Conclusions: High level of CVH, defined by LE8, was associated with significantly lower risks of CHD, stroke, and CVD. Such relationships are more pronounced among women, younger adults, and people with a lower genetic predisposition.

Background Individual unhealthy sleep behaviours have been associated with increased risks of all-cause mortality and deaths due to cardiovascular disease (CVD) or cancer. The evidence regarding the association of sleep patterns with these risks is limited. Objective To examine the associations of sleep patterns with all-cause, CVD and cancer mortality in a large prospective cohort. Methods This prospective cohort study included 283,443 adults from UK Biobank without CVD and cancer at baseline. We created a healthy sleep score and sleep patterns combining five individual sleep behaviours. Results During a mean (standard deviation) of 8.9 (1.1) years (2.5 million person-years) of follow up, a total of 7936 all-cause deaths, 762 CVD-caused deaths, and 4540 cancer-caused deaths occurred during follow up. One point increase of the healthy sleep score was associated with a 4-11% lower risk of all-cause mortality (Hazard Ratio [HR], 0.94; 95% CI, 0.92-0.96), CVD mortality (HR, 0.89; 95% CI, 0.83-0.95) and cancer mortality (HR, 0.96; 95% CI, 0.93-0.99), with adjustment for age, sex, assessment centres, smoking status, alcohol intake status, socioeconomic status and physical activity. Compared with participants with an unfavourable sleep pattern, those with a favourable sleep pattern had 24-42% lower risks of all-cause and CVD mortality. The association with all-cause mortality tended to be stronger among underweight participants and those with insufficient physical activity. Conclusions A healthy sleep pattern was associated with lower risks of all-cause mortality and mortality from CVD and cancer. Our findings highlight the importance of improving overall sleep behaviours in lowering mortality.

Background: Epigenome-wide association studies identified DNA methylation (DNAm) at several CpG sites of the carnitine palmitoyltransferase-1A (CPT1A) gene to be associated with triglycerides (TG) . However, little is known about whether the pre-treatment DNAm level at CPT1A is associated with changes in TG in response to weight-loss diet interventions. Methods: We included 673 white participants with overweight or obesity, who were randomly assigned to 1 of 4 diets varying in macronutrients components. Blood DNAm levels were profiled by a high-resolution methyl-capture sequencing at baseline. We defined the regional DNAm at CPT1A as the average methylation level over CpGs within 500 bp of the 3 TG-related DNAm sites (cg00574958, cg09737197, and cg17058475) , which were closely located. Two-year changes in TG were calculated. Linear regression models were used to examine the associations between baseline DNAm at CPT1A and 2-year changes in TG. Results: Higher regional DNAm level at CPT1A was associated with lower TG at baseline (β=-2.21, SE=1.01, p=0.029) . We found that dietary fat intake significantly modified the association between baseline DNAm at CPT1A and 2-year changes in TG, independent of concurrent weight-loss (p-int=0.018) . In the low-fat diet group, a higher regional DNAm level was associated with a greater reduction in TG at 2 years, after adjustment for age, sex, baseline body mass index, use of lipid-lowering medication, baseline TG level, and concurrent weight-loss (p=0.01) , whereas no such association was observed in the high-fat diet group (p=0.64) . A similar interaction pattern was also found between dietary fat and baseline DNAm at CPT1A on 2-year changes of TG in very-low-density lipoprotein without apolipoprotein C-III (p-int=0.01) . Conclusion: Our data indicate that participants with a higher regional DNAm level at CPT1A benefited more in long-term improvement in TG when assigned a low-fat weight-loss diet. Disclosure X.Li: None. X.Shao: None. Q.Xue: None. E.Grundberg: None. G.Bray: None. F.Sacks: None. L.Qi: None. Funding The study was supported by grants from the National Heart, Lung, and Blood Institute (HL071981, HL034594, HL126024) , the National Institute of Diabetes and Digestive and Kidney Diseases (DK115679, DK091718, DK100383) , the Fogarty International Center (TW010790) , and Tulane Research Centers of Excellence Awards. Xiang Li was the recipient of the American Heart Association Predoctoral Fellowship Award (19PRE34380036) .

Background: MicroRNA-378a (MiR-378a) has been identified as a key regulator of glucose homeostasis and emerging as a promising target for the improvement of glycemic dysregulation including type 2 diabetes. Hypothesis: We assessed associations of circulating miR-378a-5p with improvements in insulin sensitivity and glucose metabolism in a 2-year dietary intervention trial. Methods: Circulating miR-378a-5p levels were measured at baseline and 6 months among 5participants with overweight or obesity. Outcome measurements including fasting glucose, hemoglobin A1c (HbA1c) , fasting insulin, and homeostasis model assessment-of-insulin resistance (HOMA-IR) were assessed at baseline, 6 and 24 months. Results: Higher level of miR-378a-5p was associated with greater fasting glucose (p =0.031) at baseline. Greater decrease in miR-378a-5p was significantly associated with greater improvements in fasting insulin (p =0.012) and HOMA-IR (p =0.017) over 6 months. The initial (6-months) decrease of miR-378a-5p was also significantly related to long-term (24-months) improvements in glucose, insulin, and HOMA-IR (p Conclusions: In conclusion, decreased circulating miR-378a-5p levels were related to significant improvements in insulin sensitivity and glucose metabolism in response to dietary weight-loss interventions; and dietary fat intake modified such associations. Disclosure Q.Xue: None. Y.Heianza: None. X.Li: None. H.Deng: None. J.Rood: None. G.Bray: None. F.Sacks: None. L.Qi: None. Funding National Heart, Lung, and Blood Institute (HL071981, HL034594, and HL126024) ;National Institute of Diabetes and Digestive and Kidney Diseases (DK115679, DK091718, and DK100383) ;Fogarty International Center (TW010790)

Circulating microRNAs (miRNAs) have been implicated in controlling whole-body metabolism through intercellular communications. MicroRNA-193b (miR-193b) plays essential roles in white and brown adipocyte adipogenesis and regulating adipokine secretion. We tested whether changes in circulating miR-193b were associated with reductions of adiposity and various fat depots in response to weight-loss dietary and lifestyle interventions in two independent trials. This study included participants of a 2-year weight-loss diet intervention trial (POUNDS Lost) with data on changes in circulating miR-193b-5p from baseline to 6 months after interventions (n=509) . We also assessed changes in circulating miR-193b-5p over 18 months in the CENTRAL weight-loss lifestyle intervention trial (n=111) . At baseline, higher miR-193b was associated with greater adiposity, energy expenditure, ectopic fat depots, and lower adiponectin levels. In response to the interventions, decreases in miR-193b were associated with larger reductions of weight (p In conclusion, changes in circulating miR-193b in response to weight-loss diet/lifestyle interventions were significantly associated with the long-term successful weight loss and reductions of fat depots closely related to the risk of diabetes. Disclosure Y.Heianza: None. F.Sacks: None. L.Qi: None. Q.Xue: None. J.Rood: None. K.K.Krohn: None. A.Yaskolka meir: None. M.Blüher: Consultant; AstraZeneca, Lilly, Novo Nordisk A/S, Pfizer Inc., Sanofi, Speaker's Bureau; Bayer AG, Boehringer Ingelheim International GmbH, Novartis AG. P.Kovacs: None. I.Shai: None. G.Bray: None. Funding NIH (DK091718, DK100383, DK115679)

Introduction: MicroRNA-21 (MiR-21) is one of the most highly expressed microRNAs in the cardiovascular system and plays a pivotal role in the pathogenesis of cardiovascular diseases (CVD). Hypothesis: We assessed the hypothesis that circulating miR-21 is associated with the change in overall CVD risk and individual cardiovascular risk factors in response to weight-loss diet interventions. Methods: Circulating miR-21 was measured at baseline and 6 months in 511 participants with overweight or obesity from the 2-year weight-loss diet intervention study, the POUNDS LOST trial. MiR-21-5p is a mature sequence derived from the 5' end of the miR-21 stem-loop precursor and another mature sequence, miR-21-3p, is derived from the 3' end of miR-21. The primary outcome was the change in the global Framingham Risk Score (FRS), which estimates the 10-year probability of a CVD event. Secondary outcomes were the change in the FRS components including systolic blood pressure, diastolic blood pressure, total cholesterol, high-density lipoprotein cholesterol, low-density lipoprotein cholesterol, triglycerides, insulin, homeostatic model assessment of insulin resistance, gut microbiota metabolites, and adiposity measures. Results: Circulating miR-21-3p levels were positively associated with fasting insulin ( P =0.011), insulin resistance ( P =0.008), and triglycerides ( P =0.003) at baseline. We found that lower pre-intervention expression levels of miR-21-3p and miR-21-5p were related to a greater reduction in FRS (β[SE]=11.020 [5.025], P =0.029 and β[SE]=0.023 [0.008], P =0.005, respectively) over 6 months. The trajectory of the FRS across the 2-year intervention period differed between the patterns of pre-intervention miR-21 ( P time*miR-21-3p =0.002 and P time*miR-21-5p =0.033). Compared to those with a higher level of miR-21, participants with a lower level of miR-21 had a greater decrease in FRS from baseline to 6 months and then increased substantially from 6 months to 2 years. We also found significant interactions between baseline miR-21-5p and dietary fat on change in FRS ( P fat diets* miR-21-5p =0.04). Participants with a lower level of baseline miR-21-5p on the low-fat diet group showed a greater reduction of FRS than those on the high-fat diet at 6 months. In addition, a greater decrease in miR-21-3p rather than miR-21-5p was significantly associated with a greater reduction in body weight (PP =0.002), and improvement in trimethylamine N-oxide ( P =0.011) at 6 months. These associations were attenuated at 2 years of the intervention. Conclusions: In conclusion, decreased circulating miR-21 levels were related to a significant reduction in overall CVD risk and improvement in individual cardiovascular risk factors in response to dietary weight-loss interventions.

Background: MicroRNA (miRNAs) are short non-coding RNAs that play important roles in regulating gene expression. Recently, miR-128-1 was identified as a potential thrifty miRNA that controls energy expenditure, contributing to obesity and metabolic diseases. miR-128-1 is located at the locus harboring the lactase ( LCT ) gene, a hallmark gene of the positive selection and evolutionary adaptation to famine by promoting energy storage. Hypothesis: We investigated whether circulating thrifty miR-128-1 was associated with insulin resistance, adiposity, and energy expenditure among adults with overweight and obesity and tested whether miR-128-1 was associated with changes in these metabolic conditions after low-calorie weight-loss diet interventions. Methods: We included 495 participants from the POUNDS Lost trial with available data on circulating miR-128-1-5p measured at baseline and 6 months after the intervention. The LCT genotype rs4988235, which is responsible for lactase persistence in Europeans, was assessed to analyze the relationship between miR-128-1 and the positive selection at this locus. Outcome measurements included adiposity (weight, waist circumference [WC], and body composition), insulin resistance (HOMA-IR), and energy metabolism. Habitual physical activity was assessed by the Baecke questionnaires; total, leisure time, sport, work index were calculated. Results: The LCT rs4988235 (A) allele was associated with higher levels of miR-128-1 ( p =0.04). Pre-intervention miR-128-1 was significantly and positively associated with HOMA-IR ( p =0.009), insulin ( p =0.008), BMI ( p =0.0002), WC ( p =0.036), and whole-body total % fat mass ( p =0.013), and energy metabolism (resting energy expenditure, oxygen consumption, and carbon dioxide production; p p =0.001) and subscales (leisure index ( p =0.007) and sport index ( p =0.02)). We found significant interactions between miR-128-1 and physical activity on changes in adiposity and insulin resistance at 6 months after the interventions ( P interaction =0.03 for weight; P interaction =0.004 for WC; and P interaction =0.01 for HOMA-IR). Participants with lower levels of physical activity and higher miR-128-1 at baseline responded better to the intervention to improve HOMA-IR at 6 months. There were also significant interactions between changes in miR-128-1 and changes in physical activity on changes in HOMA-IR and energy expenditure. Conclusions: Elevated levels of circulating miR-128-1 were associated with greater adiposity and insulin resistance in adults with overweight and obesity. The thrifty miR-128-1 interacted with physical activity on changes in energy metabolism and insulin sensitivity in response to weight-loss diet interventions.

Background: Bile acids are synthesized from cholesterol in the liver and play pivotal roles in nutrient absorption and metabolic regulation. Altered bile acid metabolism is related to dyslipidemia, and microRNA-34a (miR-34a) may play a pivotal role in the treatment of lipid homeostasis by regulating the bile acid biosynthesis. Hypothesis: We investigated whether weight-loss diet-induced changes in primary and secondary bile acids were associated with the improvements in lipid profiles. We also tested the role of circulating miR-34a in the altered bile acid metabolism in adults with overweight and obesity. Methods: A total of 520 participants of a weight-loss dietary intervention (the POUNDS Lost trial) with data on bile acids and miR-34a were included in the present analysis. Circulating levels of miR-34a and bile acid subtypes (primary and secondary unconjugated bile acids and their taurine-/glycine-conjugates) were measured at baseline and 6 months after the intervention. Outcome measurements were improvements in lipids (triglycerides and cholesterol) in response to the interventions. Results: At baseline, higher levels of primary bile acids (chenodeoxycholate (CDCA), glycocholate (GCA), taurocholate (TCA), and glycochenodeoxycholate (GCDCA) were associated with elevated levels of triglycerides ( P FDR P FDR Conclusions: Changes in the specific primary and secondary bile acid subtypes were related to the improvements in lipid profiles in response to weight-loss diet interventions. Circulating miR-34a might in part contribute to the alterations in the primary bile acid subtypes in obese adults.

Aims We analyzed whether the frequency of adding salt to foods was associated with the hazard of premature mortality and life expectancy. Methods and results A total of 501 379 participants from UK biobank who completed the questionnaire on the frequency of adding salt to foods at baseline. The information on the frequency of adding salt to foods (do not include salt used in cooking) was collected through a touch-screen questionnaire at baseline. We found graded relationships between higher frequency of adding salt to foods and higher concentrations of spot urinary sodium or estimated 24-h sodium excretion. During a median of 9.0 years of follow-up, 18 474 premature deaths were documented. The multivariable hazard ratios [95% confidence interval (CI)] of all-cause premature mortality across the increasing frequency of adding salt to foods were 1.00 (reference), 1.02 (0.99, 1.06), 1.07 (1.02, 1.11), and 1.28 (1.20, 1.35) (P-trend Conclusions Our findings indicate that higher frequency of adding salt to foods is associated with a higher hazard of all-cause premature mortality and lower life expectancy.

Introduction: MicroRNA (miRNAs) are short non-coding RNAs that regulate gene expression at the translational level. Circulating exosomal miRNAs have emerged as novel biomarkers of obesity, insulin resistance, and related metabolic abnormalities. miR-375 has been identified as a regulator of insulin secretion, and miR-375 may also be involved in regulating visceral adiposity and ectopic fat accumulation in the liver. Hypothesis: We aimed to investigate whether changes in circulating miR-375 induced by lifestyle interventions were related to improved visceral adiposity and hepatic fat depot among individuals with abdominal obesity. Methods: This study included adults with abdominal obesity from an 18-month diet and physical activity intervention study (the CENTRAL trial) and with available data on circulating miR-375-3p at baseline (n=222) and 18 months (n=152). Magnetic resonance imaging (MRI) was performed to assess body fat distribution and ectopic fat depots. Outcome measurements included 18-month changes in abdominal (total [TAT], visceral [VAT], deep subcutaneous [DSAT], superficial subcutaneous [SSAT]) adipose tissue area and intrahepatic fat content. Results: At baseline, higher levels of miR-375 were related to lower degrees of BMI ( p =0.0006), waist circumference ( p =0.0008), as well as abdominal adipose tissue area (TAT [ p =0.0024], VAT [ p =0.059], DSAT [ p =0.011], and SSAT [ p =0.0027]). There was a significant increase in circulating miR-375 levels after the 18-month lifestyle interventions ( P by paired t-test < 0.0001). When we analyzed effects (β) of per 1 SD increase in miR-375 changes on the outcomes after adjusting for covariates including baseline levels of miR-375 and BMI, we found that greater increase in miR-375 was associated with larger reductions of VAT (β -8.09 [3.09] cm 2 , p =0.01), as well as DSAT (β -7.87 [2.21] cm 2 , p =0.001) and intrahepatic fat content (β -1.38% [0.5%], p =0.006) at 18 months. Conclusions: Circulating miR-375 levels significantly increased after the long-term lifestyle interventions, and greater increases in miR-375 were related to significant improvements in visceral adiposity and liver fat content that are closely related to insulin resistance in adults with abdominal obesity.

Introduction: Thioredoxin Interacting Protein ( TXNIP ) functions as a master regulator for glucose homeostasis. Hypomethylation at the 5'-cytosine-phosphate-guanine-3' (CpG) site cg19693031 of TXNIP has been consistently related to islet dysfunction, hyperglycemia, and type 2 diabetes. DNA methylation (DNAm) may reveal the missing mechanistic link between obesity and type 2 diabetes; however, little is known about whether the DNAm level at TXNIP in blood is associated with glycemic changes in response to weight-loss diet interventions. Hypothesis: We hypothesized that participants with varying DNAm level of TXNIP responded differentially to weight-loss diet interventions on glycemic changes. Method: We included 639 overweight and obese participants from Preventing Overweight Using Novel Dietary Strategies (POUNDS Lost) trial, who were randomly assigned to 1 of 4 weight-loss diets varying in macronutrient contents. Blood DNAm levels were profiled by a high-resolution methylC-capture sequencing at baseline. We defined regional methylation level of TXNIP as average methylation level over CpGs within 500 bp of cg19693031. Results: Higher regional methylation level at TXNIP was correlated with lower fasting glucose, HbA1c, and Homeostatic Model Assessment of Insulin Resistance (HOMA-IR) at baseline (pTXNIP showed significantly different changes in fasting insulin (p-interaction=0.007) and HOMA-IR (p-interaction=0.009) in response to average- vs. high-protein diet intervention at 6 months, independent of weight loss. Among participants with an average-protein diet, higher regional methylation level at TXNIP was associated with a greater reduction of fasting insulin and HOMA-IR at 6 months, while an opposite association was found among those with a high-protein diet. Such difference was attenuated to be nonsignificant at 2 years. Conclusions: DNAm level at TXNIP in blood may modify the effects of diet interventions on changes in glycemic traits, independent of weight loss. Overweight and obese individuals with higher methylation level at TXNIP may benefit more in improving insulin and HOMA-IR by consuming an average-protein diet than a high-protein diet.

The orexigenic gut hormone, ghrelin, has been implicated in the regulation of appetite, energy homeostasis, and adiposity. We investigated associations of plasma ghrelin changes after consuming weight loss diets with improvements in general adiposity, body fat accumulation, and energy metabolism among overweight and obese adults. We also tested whether short-term ghrelin changes in response to weight-loss diets were predictive of long-term successful weight loss. This study included 217 overweight and obese adults who participated in a weight-loss diet intervention study, the POUNDS Lost trial. Plasma ghrelin levels were measured at baseline and 6 months after the intervention. We examined associations of the 6-month changes in plasma ghrelin levels from the baseline with changes in weight, waist circumference (WC), body fat composition assessed by dual-energy X-ray absorptiometry (DEXA), fat distribution assessed by CT scans, and resting energy expenditure (REE). At baseline, higher ghrelin levels were significantly associated with lower adiposity (weight, WC, and body fat composition) and REE (p Disclosure Y. Heianza: None. Q. Xue: None. G. Bray: None. F. Sacks: None. L. Qi: None. J. Tong: None. Funding National Institute of Diabetes and Digestive and Kidney Diseases (DK091718, DK100383, DK115679)

MicroRNAs (miRNAs) are small non-coding RNAs, and adipose tissue is a major source of circulating miRNAs that may regulate body fat distribution. We investigated whether changes in circulating adipose-derived miRNAs (miR-99a-5p, miR-99b-5p, and miR-100-5p) were related to improvements in visceral and ectopic fat depots in abdominal obese adults who participated in the CENTRAL diet-and-lifestyle intervention trial. We further tested whether changes in these miRNAs were related to improved glucose metabolism and insulin sensitivity after the intervention. Plasma levels of miRNAs were measured both at baseline (n=227) and 18 months (n=157) after the intervention; changes in miRNAs from baseline to 18 months were calculated. Magnetic resonance imaging (MRI) was performed to assess visceral adipose tissue (VAT), intrahepatic fat %, and pancreatic fat %. At baseline, higher levels of miR-99a-5p and miR-100-5p were associated with greater intrahepatic fat (p Disclosure Y. Heianza: None. P. Kovacs: None. I. Shai: None. L. Qi: None. K. K. Krohn: None. M. Wang: None. A. Yaskolka meir: None. Q. Xue: None. S. Ziesche: None. U. Ceglarek: Research Support; Self; Roche Diagnostics Germany. M. Blüher: Consultant; Self; Amgen Inc., AstraZeneca, Bayer AG, Boehringer Ingelheim International GmbH, Lilly Diabetes, Novo Nordisk, Sanofi-Aventis Deutschland GmbH, Speaker’s Bureau; Self; Daiichi Sankyo, Novartis AG. M. Keller: None. Funding National Institute of Diabetes and Digestive and Kidney Diseases (DK091718, DK100383, DK115679)