Circulating serum vitamin D levels and total body bone mineral density: A Mendelian randomization study

Until recently, randomized controlled trials have not demonstrated convincing evidence that vitamin D, or vitamin D in combination with calcium supplementation could improve bone mineral density (BMD), osteoporosis and fracture. It remains unclear whether vitamin D levels are causally associated with total body BMD. Here, we performed a Mendelian randomization study to investigate the association of vitamin D levels with total body BMD using a large‐scale vitamin D genome‐wide association study (GWAS) dataset (including 79 366 individuals) and a large‐scale total body BMD GWAS dataset (including 66,628 individuals). We selected three Mendelian randomization methods including inverse‐variance weighted meta‐analysis (IVW), weighted median regression and MR‐Egger regression. All these three methods did not show statistically significant association of genetically increased vitamin D levels with total body BMD. Importantly, our findings are consistent with recent randomized clinical trials and Mendelian randomization study. In summary, we provide genetic evidence that increased vitamin D levels could not improve BMD in the general population. Hence, vitamin D supplementation alone may not be associated with reduced fracture incidence among community‐dwelling adults without known vitamin D deficiency, osteoporosis, or prior fracture.     

Causal association between heart failure and bone mineral density: Insights from a two-sample bidirectional Mendelian randomization study

In recent times, the association between HF and BMD has attracted enormous interest in the scientific community. However, published epidemiological observational studies on the relationship between HF and BMD remain inconclusive. Herein, we evaluated from the analytical perspective a two-sample bidirectional MR study to analyze the causal association between HF and BMD using a summary-level GWAS Catalog. To select instrumental SNPs strongly associated with exposure, we took a series of rigorous quality control steps at the time of analysis. The causal MR assessment of HF on the risk of BMD was performed first and then in the opposite direction. To make the conclusions more reliable and robust, the fixed-effects IVW, weighted median-based method, MR–Egger, simple mode and weighted mode were utilized. A maximum likelihood model was also used if necessary. MR–Egger regression, IVW “leave-one-out” sensitivity analysis, MR-PRESSO, MR–Egger intercept test and Cochran's Q statistic methods were used to assess heterogeneity and pleiotropy. Our MR studies supported a meaningful causal association between HF and TB-BMD (IVW: OR = 0.78, 95% CI: 0.68–0.87, p = 0.00588). At the same time, we did not find a significant causal relationship between HF and FA-BMD, FN-BMD or LS-BMD. No significant causal relationships between BMD and HF were observed. This bidirectional MR analysis suggested a causal association of HF with only low TB-BMD, while the reverse causality hypothesis was not found. Studies of the prevention and treatment of total bone mineral density decline in patients with heart failure need to be performed.     

On Mendelian randomization analysis of case-control study

Mendelian randomization (MR) analysis uses genotypes as instruments to estimate the causal effect of an exposure in the presence of unobserved confounders. The existing MR methods focus on the data generated from prospective cohort studies. We develop a procedure for studying binary outcomes under a case-control design. The proposed procedure is built upon two working models commonly used for MR analyses and adopts a quasi-empirical likelihood framework to address the ascertainment bias from case-control sampling. We derive various approaches for estimating the causal effect and hypothesis testing under the empirical likelihood framework. We conduct extensive simulation studies to evaluate the proposed methods. We find that the proposed empirical likelihood estimate is less biased than the existing estimates. Among all the approaches considered, the Lagrange multiplier (LM) test has the highest power, and the confidence intervals derived from the LM test have the most accurate coverage. We illustrate the use of our method in MR analysis of prostate cancer case-control data with vitamin D level as exposure and three single nucleotide polymorphisms as instruments.     

Mendelian randomization analysis identified genes potentially pleiotropically associated with periodontitis

ObjectiveTo prioritize genes that were pleiotropically or potentially causally associated with periodontitis.MethodsWe applied the summary data-based Mendelian randomization (SMR) method integrating genome-wide association study (GWAS) for periodontitis and expression quantitative trait loci (eQTL) data to identify genes that were pleiotropically associated with periodontitis. We performed separate SMR analysis using CAGE eQTL data and GTEx eQTL data. SMR analysis were done for participants of European and East Asian ancestries, separately.ResultsWe identified multiple genes showing pleiotropic association with periodontitis in participants of European ancestry and participants of East Asian ancestry. PDCD2 (corresponding probe: ILMN_1758915) was the top hit showing pleotropic association with periodontitis in the participants of European ancestry using CAGE eQTL data, and BX093763 (corresponding probe: ILMN_1899903) and AC104135.3 (corresponding probe: ENSG00000204792.2) were the top hits in the participants of East Asian ancestry using CAGE eQTL data and GTEx eQTL data, respectively.ConclusionWe identified multiple genes that may be involved in the pathogenesis of periodontitis in participants of European ancestry and participants of East Asian ancestry. Our findings provided important leads to a better understanding of the mechanisms underlying periodontitis and revealed potential therapeutic targets for the effective treatment of periodontitis.     

t10c12-CLA maintains higher bone mineral density during aging by modulating osteoclastogenesis and bone marrow adiposity

Conjugated linoleic acid (CLA) has been shown to positively influence calcium and bone metabolism. Earlier, we showed that CLA (equal mixture of c9t11-CLA and t10c12-CLA) could protect age-associated bone loss by modulating inflammatory markers and osteoclastogenesis. Since, c9t11-CLA and t10c12-CLA isomers differentially regulate functional parameters and gene expression in different cell types, we examined the efficacy of individual CLA isomers against age-associated bone loss using 12 months old C57BL/6 female mice fed for 6 months with 10% corn oil (CO), 9.5% CO + 0.5% c9t11-CLA, 9.5% CO + 0.5% t10c12-CLA or 9.5% CO + 0.25% c9t11-CLA + 0.25% t10c12-CLA. Mice fed a t10c12-CLA diet maintained a significantly higher bone mineral density (BMD) in femoral, tibial and lumbar regions than those fed CO and c9t11-CLA diets as measured by dual-energy-X-ray absorptiometry (DXA). The increased BMD was accompanied by a decreased production of osteoclastogenic factors, that is, RANKL, TRAP5b, TNF-alpha and IL-6 in serum. Moreover, a significant reduction of high fat diet-induced bone marrow adiposity was observed in t10c12-CLA fed mice as compared to that of CO and c9t11-CLA fed mice, as measured by Oil-Red-O staining of bone marrow sections. In addition, a significant reduction of osteoclast differentiation and bone resorbing pit formation was observed in t10c12-CLA treated RAW 264.7 cell culture stimulated with RANKL as compared to that of c9t11-CLA and linoleic acid treated cultures. In conclusion, these findings suggest that t10c12-CLA is the most potent CLA isomer and it exerts its anti-osteoporotic effect by modulating osteoclastogenesis and bone marrow adiposity.     

Cardiometabolic risk factors for COVID-19 susceptibility and severity: A Mendelian randomization analysis

BackgroundEpidemiological studies report associations of diverse cardiometabolic conditions including obesity with COVID-19 illness, but causality has not been established. We sought to evaluate the associations of 17 cardiometabolic traits with COVID-19 susceptibility and severity using 2-sample Mendelian randomization (MR) analyses.Methods and findingsWe selected genetic variants associated with each exposure, including body mass index (BMI), at p < 5 × 10⁻⁸ from genome-wide association studies (GWASs). We then calculated inverse-variance-weighted averages of variant-specific estimates using summary statistics for susceptibility and severity from the COVID-19 Host Genetics Initiative GWAS meta-analyses of population-based cohorts and hospital registries comprising individuals with self-reported or genetically inferred European ancestry. Susceptibility was defined as testing positive for COVID-19 and severity was defined as hospitalization with COVID-19 versus population controls (anyone not a case in contributing cohorts). We repeated the analysis for BMI with effect estimates from the UK Biobank and performed pairwise multivariable MR to estimate the direct effects and indirect effects of BMI through obesity-related cardiometabolic diseases. Using p < 0.05/34 tests = 0.0015 to declare statistical significance, we found a nonsignificant association of genetically higher BMI with testing positive for COVID-19 (14,134 COVID-19 cases/1,284,876 controls, p = 0.002; UK Biobank: odds ratio 1.06 [95% CI 1.02, 1.10] per kg/m²; p = 0.004]) and a statistically significant association with higher risk of COVID-19 hospitalization (6,406 hospitalized COVID-19 cases/902,088 controls, p = 4.3 × 10⁻⁵; UK Biobank: odds ratio 1.14 [95% CI 1.07, 1.21] per kg/m², p = 2.1 × 10⁻⁵). The implied direct effect of BMI was abolished upon conditioning on the effect on type 2 diabetes, coronary artery disease, stroke, and chronic kidney disease. No other cardiometabolic exposures tested were associated with a higher risk of poorer COVID-19 outcomes. Small study samples and weak genetic instruments could have limited the detection of modest associations, and pleiotropy may have biased effect estimates away from the null.ConclusionsIn this study, we found genetic evidence to support higher BMI as a causal risk factor for COVID-19 susceptibility and severity. These results raise the possibility that obesity could amplify COVID-19 disease burden independently or through its cardiometabolic consequences and suggest that targeting obesity may be a strategy to reduce the risk of severe COVID-19 outcomes.

Aaron Leong and co-workers investigate causal risk factors for COVID-10 illness and severity.     

Exercise and bone mineral density in men: a meta-analysis.

The purpose of this study was to use the meta-analytic approach to examine the effects of exercise on bone mineral density (BMD) in men. A total of 26 effect sizes (ES) representing 225 subjects from 8 studies met the criteria for inclusion. When BMD sites assessed were specific to the sites loaded during exercise, increases of approximately 2.6% (2.1% in the exercisers and -0.5% in the controls) were found. These results were statistically significant (ES = 0.213, 95% bootstrap confidence interval = 0.007-0.452). Statistically significant ES changes were found for older (>31 yr) but not younger (<31 yr) adults, with differences between groups statistically significant (P = 0.04). Statistically significant changes were also observed at the femur, lumbar, and os calcis sites. The results of this study suggest that site-specific exercise may help improve and maintain BMD at the femur, lumbar, and os calcis sites in older men. However, the biological importance of the small changes observed for most outcomes, quality of studies, and limited data pool prevent us from forming any firm conclusion regarding the use of exercise for maintaining and/or improving BMD in men. Clearly, a need exists for additional studies.     

Associations between osteoprotegerin polymorphisms and bone mineral density: a meta-analysis

Associations between polymorphisms of the osteoprotegerin gene (OPG) and bone mineral density (BMD) have been studied by several research groups, but results are mixed. Accordingly, the authors performed a meta-analysis on studies of associations between OPG polymorphisms and BMD. Appropriate studies were identified using MEDLINE and by manual searching. A total of eight separate comparisons were considered in this meta-analysis. Individuals with the GG genotype of G1181C were found to have a significantly lower mean lumbar BMD than subjects with the CC genotype (WMDs −0.051 g/cm², 95% confidence interval −0.079−−0.023, P < 0.001), and similar results were obtained in European and Asian populations. In contrast to G1181C, no association was found between the A163G and T950C polymorphisms and lumbar BMD. In terms of femoral neck BMD, the GG genotype of G1181C was associated with a significantly lower BMD than the CC genotype in Europeans but not in Asians. Total hip BMD was lower for the GG genotype of G1181C than for the CC or GC genotypes in Europeans. A difference in total hip BMD was found between the AG and GG genotypes of the A163G polymorphism by meta-analyses in Europeans, but no differences were found between the genotypes of the T950C polymorphism and total hip BMD in Europeans. Summarizing, the present study demonstrates that the OPG G1181C polymorphism is associated with lumbar BMD in Europeans and Asians, and with femoral neck and total hip BMD in Europeans only.     

Alcohol intake and blood pressure: a systematic review implementing a Mendelian randomization approach

Background

Alcohol has been reported to be a common and modifiable risk factor for hypertension. However, observational studies are subject to confounding by other behavioural and sociodemographic factors, while clinical trials are difficult to implement and have limited follow-up time. Mendelian randomization can provide robust evidence on the nature of this association by use of a common polymorphism in aldehyde dehydrogenase 2 (ALDH2) as a surrogate for measuring alcohol consumption. ALDH2 encodes a major enzyme involved in alcohol metabolism. Individuals homozygous for the null variant (*2*2) experience adverse symptoms when drinking alcohol and consequently drink considerably less alcohol than wild-type homozygotes (*1*1) or heterozygotes. We hypothesise that this polymorphism may influence the risk of hypertension by affecting alcohol drinking behaviour.

Methods and Findings

We carried out fixed effect meta-analyses of the ALDH2 genotype with blood pressure (five studies, n = 7,658) and hypertension (three studies, n = 4,219) using studies identified via systematic review. In males, we obtained an overall odds ratio of 2.42 (95% confidence interval [CI] 1.66–3.55, p = 4.8 × 10⁻⁶) for hypertension comparing *1*1 with *2*2 homozygotes and an odds ratio of 1.72 (95% CI 1.17–2.52, p = 0.006) comparing heterozygotes (surrogate for moderate drinkers) with *2*2 homozygotes. Systolic blood pressure was 7.44 mmHg (95% CI 5.39–9.49, p = 1.1 × 10⁻¹²) greater among *1*1 than among *2*2 homozygotes, and 4.24 mmHg (95% CI 2.18–6.31, p = 0.00005) greater among heterozygotes than among *2*2 homozygotes.

Conclusions

These findings support the hypothesis that alcohol intake has a marked effect on blood pressure and the risk of hypertension.     

Volumetric and projective bone mineral density

Projectional bone mineral density measurement so far used extensively in radiogrammetry and single and dual source absorptiometry is confronted with a serious limitation for the accurate evaluation of true density artifactually providing higher values along with the increase of body size and bone depth on account of the omission of one dimension. Computed tomography is capable of measuring true volumetric density and also accomplishes a separate measurement of trabecular and cortical bone especially on application to the distal and mid-radius with abundant cortical bone in peripheral computed tomography (pQCT). New lines of information may be obtained by such separate trabecular and cortical bone measurement in decreases of bone density due to various causes, estrogen withdrawal, corticosteroid, diabetes mellitus, renal failure, etc. Dynamic analysis of the result of pQCT may also make it possible to assess bone strength and resistance to fracture.     

Anorexia nervosa: hypogonadotrophic hypogonadism and bone mineral density

Anorexia nervosa is a chronic illness that involves a reduction in caloric intake, loss of weight and amenorrhoea, either primary or secondary. In addition to prolonged amenorrhoea, osteopenia and osteoporosis are the most frequent complications. Patients exhibit an alteration in the hypothalamic-pituitary-gonadal axis, which is responsible for the menstrual disorders. The increase in gonadotrophin secretion that can be observed after ponderal recuperation suggests that malnutrition could be the most important mechanism involved in the decrease in gonadotrophin secretion. The loss of fat tissue, as a consequence of the restriction of nutrients, has been associated with hypoleptinaemia, abnormal secretion of peptides implicated in food control (neuropeptide Y, melanocortins and corticotrophin-releasing hormone, among others) and diminution of the amount of total body fat. Despite oestrogen therapy, the severe loss of bone mass may progress. Other factors such as weight loss, duration of amenorrhoea and low insulin-like growth factor-I (IGF-I) levels could contribute to the loss of bone mass in women with anorexia nervosa. The recuperation of weight and, in particular, the amount of total body fat could lead to the spontaneous recuperation of menstruation.     

Pathway analysis of genome-wide association study for bone mineral density

The aim of this study was to identify the candidate causal single nucleotide polymorphisms (SNPs) and candidate causal mechanisms that contribute to bone mineral density (BMD) and to generate a SNP to gene to pathway hypothesis using an analytical pathway-based approach. We used hip BMD GWAS data of the genotypes of 301,019 SNPs in 5,715 Europeans. ICSNPathway (identify candidate causal SNPs and pathways) analysis was applied to the BMD GWAS dataset. The first stage involved the pre-selection of candidate causal SNPs by linkage disequilibrium analysis and the functional SNP annotation of the most significant SNPs found. The second stage involved the annotation of biological mechanisms for the pre-selected candidate causal SNPs using improved-gene set enrichment analysis. ICSNPathway analysis identified seven candidate SNPs, eight candidate pathways, and seven hypothetical biological mechanisms. Eight pathways are as follows; gamma-hexachlorocyclohexane degradation (nominal p-value < 0.001, false discovery rate (FDR) <0.001), regulation of the smoothened signaling pathway (nominal p-value < 0.001, FDR = 0.016), TACI and BCMA stimulation of B cell immune response (nominal p-value < 0.001, FDR = 0.021), endonuclease activity (nominal p-value = 0.001, FDR = 0,026), regulation of defense response to virus (nominal p-value = 0.001, FDR = 0.028), serine_type_endopeptidase_inhibitor_activity (nominal p-value = 0.001, FDR = 0.044), endoribonuclease activity (nominal p-value = 0.002, FDR = 0.045), and myeloid leukocyte differentiation (nominal p-value = 0.001, FDR = 0.050). The most significant causal pathway was gamma-hexachlorocyclohexane degradation. CYP3A5, PON2, PON3, CMBL, PON1, ALPL, CYP3A43, CYP3A7, ACP6, ACPP, and ALPI (p < 0.05) are involved in the pathway of gamma-hexachlorocyclohexane degradation. Further examination of the gene contents revealed that DBR1, DICER1, EXO1, FEN1, POP1, POP4, RPP30, and RPP38 were involved in 2 of the 8 pathways (p < 0.05). By applying ICSNPathway analysis to BMD GWAS data, we identified seven candidate SNPs and eight pathways involving gamma-hexachlorocyclohexane degradation, which may contribute to low BMD.     

Mendelian randomization studies on atherosclerotic cardiovascular disease: evidence and limitations

Epidemiological research has revealed a galaxy of biomarkers, such as genes, molecules or traits, which are associated with increased risk of atherosclerotic cardiovascular diseases(ASCVD). However, the etiological basis remains poorly characterized.Mendelian randomization(MR) involves the use of observational genetic data to ascertain the roles of disease-associated risk factors and, in particular, differentiate those reflecting the presence or severity of a disease from those contributing causally to a disease. Over the past decade, MR has evolved into a fruitful approach to clarifying the causal relation of a biomarker with ASCVD and to verifying potential therapeutic targets for ASCVD. In this review, we selected high-quality MR studies on ASCVD, examined the causal relationship of a series of biomarkers with ASCVD, and elucidated the role of MR in validating biomarkers as a therapeutic target by comparing the results from MR studies and randomized clinical trials(RCTs) for the treatment of ASCVD. The good agreement between the results derived by MR and RCTs suggests that MR could be performed as a screening process before novel drug development. However, when designing and interpreting a MR study, the assumptions and limitations inherent in this approach should be taken into account. Novel methodological developments, such as sensitivity analysis, will help to strengthen the validity of MR studies.     

Stature and long-term labor market outcomes: Evidence using Mendelian randomization

We use the Young Finns Study (N = ∼2000) on the measured height linked to register-based long-term labor market outcomes. The data contain six age cohorts (ages 3, 6, 9, 12, 15 and 18, in 1980) with the average age of 31.7, in 2001, and with the female share of 54.7. We find that taller people earn higher earnings according to the ordinary least squares (OLS) estimation. The OLS models show that 10 cm of extra height is associated with 13% higher earnings. We use Mendelian randomization, with the genetic score as an instrumental variable (IV) for height to account for potential confounders that are related to socioeconomic background, early life conditions and parental investments, which are otherwise very difficult to fully account for when using covariates in observational studies. The IV point estimate is much lower and not statistically significant, suggesting that the OLS estimation provides an upward biased estimate for the height premium. Our results show the potential value of using genetic information to gain new insights into the determinants of long-term labor market success.     

The bone behind a low areal bone mineral density: peripheral quantitative computed tomographic analysis in a woman with osteogenesis imperfecta

Areal bone mineral density (BMD) is the most widely used densitometric parameter. However, this approach makes it difficult to understand the structural basis of bone diseases, because a large number of bone properties are integrated into a single number. This is exemplified in the present case of a 27-year-old woman with osteogenesis imperfecta type I. Peripheral quantitative computed tomographic analysis at the radial metaphysis and at the radial diaphysis revealed a decreased areal BMD at both sites (z score -3.9 and -3.4, respectively). Yet, the structural basis for this decrease was different for the two locations: At the distal radius areal BMD was decreased because volumetric BMD was very low, whereas bone size was above the mean of the reference range. At the proximal radius areal BMD was decreased, because bone size was very low but volumetric BMD was above average. Bone mineral content of the radial diaphysis was very low for forearm muscle size, a finding which is compatible with Frost's hypothesis that the mechanostat setpoint is increased in osteogenesis imperfecta.