Rare and low-frequency coding variants alter human adult height.

Height is a highly heritable, classic polygenic trait with approximately 700 common associated variants identified through genome-wide association studies so far. Here, we report 83 height-associated coding variants with lower minor-allele frequencies (in the range of 0.1-4.8%) and effects of up to 2 centimetres per allele (such as those in IHH, STC2, AR and CRISPLD2), greater than ten times the average effect of common variants. In functional follow-up studies, rare height-increasing alleles of STC2 (giving an increase of 1-2 centimetres per allele) compromised proteolytic inhibition of PAPP-A and increased cleavage of IGFBP-4 in vitro, resulting in higher bioavailability of insulin-like growth factors. These 83 height-associated variants overlap genes that are mutated in monogenic growth disorders and highlight new biological candidates (such as ADAMTS3, IL11RA and NOX4) and pathways (such as proteoglycan and glycosaminoglycan synthesis) involved in growth. Our results demonstrate that sufficiently large sample sizes can uncover rare and low-frequency variants of moderate-to-large effect associated with polygenic human phenotypes, and that these variants implicate relevant genes and pathways.

Personal exposure to household air pollution and lung function in rural Bangladesh: A population-based cross-sectional study.

We assessed whether personal exposure to household air pollution [PM2.5 and black carbon (BC)] is associated with lung functions (FEV1, FVC, and their ratio) in non-smoking adults in rural Bangladesh. We measured personal exposure to PM2.5 using gravimetric analysis of PM2.5 mass and BC by reflectance measurement between April 2016 and June 2019. The average 24-hour PM2.5 and BC concentration was 141.0µgm-3 and 13.8µgm-3 for females, and 91.7 µgm-3 and 10.1 µgm-3 for males, respectively. A 1 µgm-3 increase in PM2.5 resulted in a 0.02 ml reduction in FEV1, 0.43 ml reduction in FVC, and 0.004% reduction in FEV1/FVC. We also found a similar inverse relationship between BC and lung functions (9.6 ml decrease in FEV1 and 18.5 ml decrease in FVC per 1µgm-3 increase in BC). A higher proportion of non-smoking biomass fuel users (50.1% of the females and 46.7% of the males) had restrictive patterns of lung function abnormalities, which need further exploration.

The Bangladesh Risk of Acute Vascular Events (BRAVE) Study: objectives and design.

During recent decades, Bangladesh has experienced a rapid epidemiological transition from communicable to non-communicable diseases. Coronary heart disease (CHD), with myocardial infarction (MI) as its main manifestation, is a major cause of death in the country. However, there is limited reliable evidence about its determinants in this population. The Bangladesh Risk of Acute Vascular Events (BRAVE) study is an epidemiological bioresource established to examine environmental, genetic, lifestyle and biochemical determinants of CHD among the Bangladeshi population. By early 2015, the ongoing BRAVE study had recruited over 5000 confirmed first-ever MI cases, and over 5000 controls "frequency-matched" by age and sex. For each participant, information has been recorded on demographic factors, lifestyle, socioeconomic, clinical, and anthropometric characteristics. A 12-lead electrocardiogram has been recorded. Biological samples have been collected and stored, including extracted DNA, plasma, serum and whole blood. Additionally, for the 3000 cases and 3000 controls initially recruited, genotyping has been done using the CardioMetabochip+ and the Exome+ arrays. The mean age (standard deviation) of MI cases is 53 (10) years, with 88 % of cases being male and 46 % aged 50 years or younger. The median interval between reported onset of symptoms and hospital admission is 5 h. Initial analyses indicate that Bangladeshis are genetically distinct from major non-South Asian ethnicities, as well as distinct from other South Asian ethnicities. The BRAVE study is well-placed to serve as a powerful resource to investigate current and future hypotheses relating to environmental, biochemical and genetic causes of CHD in an important but under-studied South Asian population.

Prevalence and Determinants of Chronic Obstructive Pulmonary Disease (COPD) in Bangladesh.

RATIONALE: There is a paucity of population-based data on COPD prevalence and its determinants in Bangladesh. OBJECTIVE: To measure COPD prevalence and socioeconomic and lifestyle determinants among ≥40 years Bangladeshi adults. METHODS: In a cross-sectional study, we measured lung function of 3744 randomly selected adults ≥40 years from rural and urban areas in Bangladesh, using a handheld spirometer. COPD was defined according to the Global Initiative for Chronic Obstructive Lung Disease (GOLD) criteria as post-bronchodilator ratio of Forced Expiratory Volume in 1st second (FEV1) to Forced Vital Capacity (FVC) < 0.7. In addition, COPD was also assessed by the lower limit of normal (LLN) threshold defined as lower fifth percentile for the predicted FEV1/FVC. RESULTS: The prevalence of COPD was 13.5% by GOLD criteria and 10.3% by LLN criteria. Prevalence of COPD was higher among rural than urban residents and in males than females. More than half of the COPD cases were stage II COPD by both criteria. Milder cases (Stages I and II) were over estimated by the GOLD fixed criteria, but more severe cases (Stages III and IV) were similarly classified. In multiple logistic regression analysis, older age, male sex, illiteracy, underweight, history of smoking (both current and former), history of asthma and solid fuel use were significant predictors of COPD. CONCLUSION: COPD is a highly prevalent and grossly underdiagnosed public health problem in Bangladeshi adults aged 40 years or older. Illiteracy, smoking and biomass fuel burning are modifiable determinants of COPD.

High prevalence of chronic kidney disease in a community survey of urban Bangladeshis: a cross-sectional study.

BACKGROUND: The burden of chronic kidney disease (CKD) will rise in parallel with the growing prevalence of type two diabetes mellitus in South Asia but is understudied. Using a cross-sectional survey of adults living in a middle-income neighborhood of Dhaka, Bangladesh, we tested the hypothesis that the prevalence of CKD in this group would approach that of the U.S. and would be strongly associated with insulin resistance. METHODS: We enrolled 402 eligible adults (>30 years old) after performing a multi-stage random selection procedure. We administered a questionnaire, and collected fasting serum samples and urine samples. We used the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation to estimate glomerular filtration rate, and sex-specific cut offs for albuminuria: > 1.9 mg/mmol (17 mg/g) for men, and >2.8 mg/mmol (25 mg/g) for women. We assessed health-related quality of life using the Medical Outcomes Study Short Form-12 (SF-12). RESULTS: A total of 357 (89%) participants with serum samples comprised the analytic cohort. Mean age of was 49.5 (± 12.7) years. Chronic kidney disease was evident in 94 (26%). Of the participants with CKD, 58 (62%) had albuminuria only. A participant with insulin resistance had a 3.6-fold increase in odds of CKD (95% confidence interval 2.1 to 6.4). Participants with stage three or more advanced CKD reported a decrement in the Physical Health Composite score of the SF-12, compared with participants without CKD. CONCLUSION: We found an alarmingly high prevalence of CKD--particularly CKD associated with insulin resistance-in middle-income, urban Bangladeshis.

Obesity and overweight in Bangladeshi children and adolescents: a scoping review.

BACKGROUND: Obesity and overweight in children and adolescents is an emerging public health concern alongside under-nutrition in low and middle income countries. Our aim was to conduct a scoping review of literature to ascertain what is known about childhood and adolescent overweight and/or obesity in Bangladesh. METHOD: Using the scoping review based on York methodology, a comprehensive search of published academic articles, conference proceedings and grey literature was carried out through PubMed, BanglaJOL, Google and Google scholar limited to English-written papers. We summarized prevalence, risk factors and health outcomes of obesity/overweight in young children and adolescents aged between 0 to 19 years old in Bangladesh and highlighted use of different reference standards to measure childhood obesity. RESULTS: In total 21 studies met the inclusion criteria. Nearly all of the reviewed articles used data from cross sectional studies, while only two used case-control design. Overall thirteen studies (62%) were primary research and eight (38%) included secondary data. Studies indicated an increasing trend in childhood obesity over time. Prevalence ranged from less than 1% to 17.9% based on different reference standards, with higher percentage amongst urban children across different age groups and sexes. CONCLUSION: This review demonstrated paucity of comprehensive literature on childhood obesity in Bangladesh, which might be explored through population-based prospective studies based on strong methodology and uniform reference standards. Sustainable and scalable preventative measures targeting high risk groups are required to avoid further rise.

Consequences of hypertension and chronic obstructive pulmonary disease, healthcare-seeking behaviors of patients, and responses of the health system: a population-based cross-sectional study in Bangladesh.

BACKGROUND: Non-communicable diseases are a threat to human health and economic development of low-income countries. Hypertension (HT) and chronic obstructive pulmonary disease (COPD) are two major causes of deaths, worldwide. This study assesses the health status, health-care seeking, and health provider responses among patients with these conditions. METHODS: The study carried out population-based cross-sectional survey in a rural and an urban surveillance area in Bangladesh. It interviewed all patients identified with HT and COPD at home using a structured questionnaire on the health consequences, healthcare-seeking behaviours, and coping strategies. Qualitative techniques identified key factors relating to the behaviours of patients and providers. RESULTS: COPD and HT correlate with lower activities of daily living (ADL) scores. The odds ratio (OR) for ADL scores in the combied conditions are high (OR: 3.04, p < 0.05) as compared to hypertension. Financial crises occur significantly more frequently among COPD patients in the urban site as compared to those in rural ares (12.5% vs. 2.4%, p < 0.01). Self-treatment at the onset is common. Seeking care from trained providers is higher in urban settings and is higher for HT. Referral for both COPD and hypertension was inadequate until the disease severity increased. CONCLUSIONS: COPD and HT significantly are associated with lower ADL scores and financial problems. Public-sector primary healthcare facilities should be better organised to address both conditions with the aim to reduce household poverty.

Smoking-attributable mortality in Bangladesh: proportional mortality study.

OBJECTIVE: To directly estimate how much smoking contributes to cause-specific mortality in Bangladesh. METHODS: A case-control study was conducted with surveillance data from Matlab, a rural subdistrict. Cases (n = 2213) and controls (n = 261) were men aged 25 to 69 years who had died between 2003 and 2010 from smoking-related and non-smoking-related causes, respectively. Cause-specific odds ratios (ORs) were calculated for "ever-smokers" versus "never-smokers", with adjustment for education, tobacco chewing status and age. Smoking-attributable deaths among cases, national attributable fractions and cumulative probability of surviving from 25 to 69 years of age among ever-smokers and never-smokers were also calculated. FINDINGS: The fraction of ever-smokers was about 84% among cases and 73% among controls (OR: 1.7; 99% confidence interval, CI: 1.1-2.5). ORs were highest for cancers and lower for respiratory, vascular and other diseases. A dose-response relationship was noted between age at smoking initiation and daily number of cigarettes or bidis smoked and the risk of death. Among 25-year-old Bangladeshi men, 32% of ever-smokers will die before reaching 70 years of age, compared with 19% of never-smokers. In 2010, about 25% of all deaths observed in Bangladeshi men aged 25 to 69 years (i.e. 42,000 deaths) were attributable to smoking. CONCLUSION: Smoking causes about 25% of all deaths in Bangladeshi men aged 25 to 69 years and an average loss of seven years of life per smoker. Without a substantial increase in smoking cessation rates, which are low among Bangladeshi men, smoking-attributable deaths in Bangladesh are likely to increase.

Zinc treatment for 5 or 10 days is equally efficacious in preventing diarrhea in the subsequent 3 months among Bangladeshi children.

We conducted a randomized, double-blind placebo controlled, community trial in rural Bangladesh in children 4-59 mo of age to compare the efficacy of a 5- and 10-d course of zinc therapy on the incidence and duration of diarrhea over the subsequent 90-d follow-up after initial treatment for an acute childhood diarrheal (ACD) episode. Children (n = 1622) with ACD were randomly allocated to either 5 or 10 d of zinc treatment. Female field workers visited each child daily, supervised the administration of zinc, recorded the duration of current episode, and the occurrence and duration of diarrhea over the subsequent 3 mo. The incidence of diarrhea over the 90 d of follow-up did not differ between the 5-d (1.08 ± 1.38 episodes) and 10-d (1.02 ± 1.35 episodes) groups (P = 0.35). Children in both groups experienced a comparable duration of diarrheal episodes (3.1 ± 5.6 d vs. 2.9 ± 5.6 d, 5-d vs. 10-d, respectively; P = 0.64) with a mean difference between groups within the defined range of equivalence. Time to onset of the first episode and the proportion children experiencing diarrhea during the 90-d follow-up also did not differ between groups. These findings suggest that among Bangladeshi children, a 5-d zinc treatment for ACD is as efficacious as 10 d in preventing diarrhea in the subsequent 3 mo.

Exposure to household air pollutants and endothelial dysfunction in rural Bangladesh: A cross-sectional study.

More than one third of world's population use biomass fuel for cooking that has been linked to an array of adverse health hazards including cardiovascular mortality and morbidity. As part of Bangladesh Global Environmental and Occupational Health (GEO Health) project, we assessed whether household air pollution (HAP) was associated with dysfunction in microvascular circulation (measured by reactive hyperemia index [RHI]). METHODS: We measured exposure to HAP (particulate matter [PM2.5], carbon monoxide [CO], and black carbon [BC]) for 48 hours of 200 healthy nonsmoker adult females who used biomass fuel for cooking. Exposure to PM2.5 and BC were measured using personal monitor, RTI MicroPEM (RTI International, NC) with an internal filter that had been both pre- and post-weighed to capture the deposited pollutants concentration. Lascar CO logger was used to measure CO. Endothelial function was measured by forearm blood flow dilatation response to brachial artery occlusion using RHI based on peripheral artery tonometry. A low RHI score (<1.67) indicates impaired endothelial function. RESULTS: Average 48 hours personal exposure to PM2.5 and BC were 144.15 µg/m3 (SD 61.26) and 6.35 µg/m3 (SD 2.18), respectively. Interquartile range for CO was 0.73 ppm (0.62-1.35 ppm). Mean logarithm of RHI (LnRHI) was 0.57 in current data. No statistically significant association was observed for LnRHI with PM2.5 (odds ratio [OR] = 0.97; 95% confidence interval [CI] = 0.92, 1.01; P = 0.16), BC (OR = 0.85; 95% CI = 0.72, 1.01; P = 0.07), and CO (OR = 0.89; 95% CI = 0.64, 1.25; P = 0.53) after adjusting for potential covariates. CONCLUSIONS: In conclusion, HAP was not associated with endothelial dysfunction among nonsmoking females in rural Bangladesh who used biomass fuel for cooking for years.

Effect of dietary fat supplementation during late pregnancy and first six months of lactation on maternal and infant vitamin A status in rural Bangladesh.

Dietary fat intake is extremely low in most communities with vitamin A deficiency. However, its role in vitamin A status of pregnant and lactating women is poorly understood. The aim of the study was to examine the effect of supplementing women with fat from mid-/late pregnancy until six months postpartum on their vitamin A status and that of their infants. Women recruited at 5-7 months of gestation were supplemented daily with 20 mL of soybean-oil (n = 248) until six months postpartum or received no supplement (n = 251). Dietary fat intake was assessed by 24-hour dietary recall at enrollment and at 1, 3 and 6 months postpartum. Concentrations of maternal plasma retinol, beta-carotene, and lutein were measured at enrollment and at 1, 3 and 6 months postpartum, and those of infants at six months postpartum. Concentration of breastmilk retinol was measured at 1, 3 and 6 months postpartum. The change in concentration of plasma retinol at three months postpartum compared to pregnancy was significantly higher in the supplemented compared to the control women (+0.04 vs -0.07 micromol/L respectively; p < 0.05). Concentrations of plasma beta-carotene and lutein declined in both the groups during the postpartum period but the decline was significantly less in the supplemented than in the control women at one month (beta-carotene -0.07 vs -0.13 micromol/L, p < 0.05); lutein -0.26 vs -0.49 micromol/L, p < 0.05) and three months (beta-carotene -0.04 vs -0.08 micromol/L, p < 0.05; lutein -0.31 vs -0.47 micromol/L, p < 0.05). Concentration of breastmilk retinol was also significantly greater in the supplemented group at three months postpartum than in the controls (0.68 +/- 0.35 vs 0.55 +/- 0.34 micromol/L respectively, p < 0.03). Concentrations of infants' plasma retinol, beta-carotene, and lutein, measured at six months of age, did not differ between the groups. Fat supplementation during pregnancy and lactation in women with a very low intake of dietary fat has beneficial effects on maternal postpartum vitamin A status.

Maternal, infant, and household factors are associated with breast-feeding trajectories during infants' first 6 months of life in Matlab, Bangladesh.

Women's breast-feeding patterns are complex, and existing definitions of breast-feeding behavior do not capture this complexity adequately. We used results from a prior qualitative study to define trajectories for feeding during the first half of infancy, and then examined household-, maternal-, and infant-level determinants of these trajectories using logistic regression analysis. The 1472 women in the study cohort lived in rural Bangladesh and were participants in the Maternal and Infant Nutrition Intervention in Matlab trial. The 3 infant feeding trajectories included women who fed only breast milk and water [full breast-feeding trajectory (FBT)]; offered mixed feeding continuously when their babies were 0-4 mo old [continuous mixed feeding trajectory (CMFT)]; and practiced any other type of breast-feeding [intermittent feeding trajectory (IFT)], which was the normative feeding behavior in this community. In adjusted regression models, women who lived in rural areas [odds ratio (OR), 2.1; 95% CI, 1.2, 3.4], came from the poorest households (OR, 3.4; 95% CI, 1.5, 7.7), and offered prelacteal (OR, 1.7; 95% CI, 1.0-2.7) were more likely to be in the FBT. Women from the richest households (OR, 1.6; 95% CI, 1.1, 2.6), employed mothers (OR, 1.7; 95% CI, 1.1, 2.6), and older mothers (OR, 1.02; 95% CI, 1.0, 1.1) were more likely to be in the CMFT, and women with higher birth-weight infants (OR, 0.6; 95% CI, 0.4-0.8) were less likely to be in the CMFT. Thus, these trajectories were associated with distinct groups of women and these results provide information useful for developing interventions to improve breast-feeding practices.

Household food security is associated with growth of infants and young children in rural Bangladesh.

OBJECTIVE: Despite a strong relationship between household food security and the health and nutritional status of adults and older children, the association of household food security with the growth of infants and young children has not been adequately studied, particularly in developing countries. We examined the association between household food security and subsequent growth of infants and young children in rural Bangladesh. DESIGN: We followed 1343 children from birth to 24 months of age who were born in the Maternal and Infant Nutrition Intervention in Matlab (MINIMat) study in rural Bangladesh. A food security scale was created from data collected on household food security from the mothers during pregnancy. Data on weight and length were collected monthly in the first year and quarterly in the second year of life. Anthropometric indices were calculated relative to the 2006 WHO child growth standards. Growth trajectories were modelled using multilevel models for change controlling for possible confounders. RESULTS: Household food security was associated (P < 0.05) with greater subsequent weight and length gain in this cohort. Attained weight, length and anthropometric indices from birth to 24 months were higher (P < 0.001) among those who were in food-secure households. Proportions of underweight and stunting were significantly (P < 0.05) lower in food-secure households. CONCLUSIONS: These results suggest that household food security is a determinant of child growth in rural Bangladesh, and that it may be necessary to ensure food security of these poor rural households to prevent highly prevalent undernutrition in this population and in similar settings elsewhere in the world.

Use of the new World Health Organization child growth standards to describe longitudinal growth of breastfed rural Bangladeshi infants and young children.

BACKGROUND: Although the National Center for Health Statistics (NCHS) reference has been widely used, in 2006 the World Health Organization (WHO) released new standards for assessing growth of infants and children worldwide. OBJECTIVE: To assess and compare the growth of breastfed rural Bangladeshi infants and young children based on the new WHO child growth standards and the NCHS reference. METHODS: We followed 1343 children in the Maternal and Infant Nutrition Intervention in Matlab (MINIMat) study from birth to 24 months of age. Weights and lengths of the children were measured monthly during infancy and quarterly in the second year of life. Anthropometric indices were calculated using both WHO standards and the NCHS reference. The growth pattern and estimates of undernutrition based on the WHO standards and the NCHS reference were compared. RESULTS: The mean birthweight was 2697 +/- 401 g, with 30% weighing <2500 g. The growth pattern of the MINIMat children more closely tracked the WHO standards than it did the NCHS reference. The rates of stunting based on the WHO standards were higher than the rates based on the NCHS reference throughout the first 24 months. The rates of underweight and wasting based on the WHO standards were significantly different from those based on the NCHS reference. CONCLUSIONS: This comparison confirms that use of the NCHS reference misidentifies undernutrition and the timing of growth faltering in infants and young children, which was a key rationale for constructing the new WHO standards. The new WHO child growth standards provide a benchmark for assessing the growth of breastfed infants and children.

Protein-coding variants implicate novel genes related to lipid homeostasis contributing to body-fat distribution.

Body-fat distribution is a risk factor for adverse cardiovascular health consequences. We analyzed the association of body-fat distribution, assessed by waist-to-hip ratio adjusted for body mass index, with 228,985 predicted coding and splice site variants available on exome arrays in up to 344,369 individuals from five major ancestries (discovery) and 132,177 European-ancestry individuals (validation). We identified 15 common (minor allele frequency, MAF ≥5%) and nine low-frequency or rare (MAF <5%) coding novel variants. Pathway/gene set enrichment analyses identified lipid particle, adiponectin, abnormal white adipose tissue physiology and bone development and morphology as important contributors to fat distribution, while cross-trait associations highlight cardiometabolic traits. In functional follow-up analyses, specifically in Drosophila RNAi-knockdowns, we observed a significant increase in the total body triglyceride levels for two genes (DNAH10 and PLXND1). We implicate novel genes in fat distribution, stressing the importance of interrogating low-frequency and protein-coding variants.

Publisher Correction: Protein-altering variants associated with body mass index implicate pathways that control energy intake and expenditure in obesity.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Household food security is associated with infant feeding practices in rural Bangladesh.

Although household food security (HHFS) has been shown to affect diet, nutrition, and health of adults and also learning in children, no study has examined associations with infant feeding practices (IFP). We studied 1343 infants born between May 2002 and December 2003 in the Maternal and Infant Nutrition Intervention in Matlab study to investigate the effect of HHFS on IFP in rural Bangladesh. We measured HHFS using a previously developed 11-item scale. Cumulative and current infant feeding scales were created from monthly infant feeding data for the age groups of 1-3, 1-6, 1-9, and 1-12 mo based on comparison to infant feeding recommendations. We used lagged, dynamic, and difference longitudinal regression models adjusting for various infant and maternal variables to examine the association between HHFS and changes in IFP, and Cox proportional hazards models to examine the influence of HHFS on the duration of breast-feeding and the time of introduction of complementary foods. Better HHFS status was associated with poor IFP during 3-6 mo but was associated with better IFP during 6-9 and 9-12 mo of age. Although better HHFS was not associated with the time of introduction of complementary foods, it was associated with the type of complementary foods given to the infants. Intervention programs to support proper IFP should target mothers in food-secure households when their babies are 3-6 mo old and also mothers in food-insecure households during the 2nd half of infancy. Our results provide strong evidence that HHFS influences IFP in rural Bangladesh.

Publisher Correction: Protein-altering variants associated with body mass index implicate pathways that control energy intake and expenditure in obesity.

In the published version of this paper, the name of author Emanuele Di Angelantonio was misspelled. This error has now been corrected in the HTML and PDF versions of the article.

Publisher Correction: Protein-altering variants associated with body mass index implicate pathways that control energy intake and expenditure in obesity.

In the version of this article originally published, one of the two authors with the name Wei Zhao was omitted from the author list and the affiliations for both authors were assigned to the single Wei Zhao in the author list. In addition, the ORCID for Wei Zhao (Department of Biostatistics and Epidemiology, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA) was incorrectly assigned to author Wei Zhou. The errors have been corrected in the HTML and PDF versions of the article.

Protein-altering variants associated with body mass index implicate pathways that control energy intake and expenditure in obesity.

Genome-wide association studies (GWAS) have identified >250 loci for body mass index (BMI), implicating pathways related to neuronal biology. Most GWAS loci represent clusters of common, noncoding variants from which pinpointing causal genes remains challenging. Here we combined data from 718,734 individuals to discover rare and low-frequency (minor allele frequency (MAF) < 5%) coding variants associated with BMI. We identified 14 coding variants in 13 genes, of which 8 variants were in genes (ZBTB7B, ACHE, RAPGEF3, RAB21, ZFHX3, ENTPD6, ZFR2 and ZNF169) newly implicated in human obesity, 2 variants were in genes (MC4R and KSR2) previously observed to be mutated in extreme obesity and 2 variants were in GIPR. The effect sizes of rare variants are ~10 times larger than those of common variants, with the largest effect observed in carriers of an MC4R mutation introducing a stop codon (p.Tyr35Ter, MAF = 0.01%), who weighed ~7 kg more than non-carriers. Pathway analyses based on the variants associated with BMI confirm enrichment of neuronal genes and provide new evidence for adipocyte and energy expenditure biology, widening the potential of genetically supported therapeutic targets in obesity.