The causal role of gut microbiota in development of osteoarthritis.

OBJECTIVE: There is considerable evidence for relationship between gut microbiota and osteoarthritis (OA), but no studies have investigated their causal relationship. METHOD: This study utilized large-scale genome-wide association studies (GWAS) summary statistics to evaluate the causal association between gut microbiota and OA risk. Specifically, two-sample Mendelian randomization (MR) approach was used to identify the causal microbial taxa for OA. Comprehensively sensitive analyses were performed to validate the robustness of results and novel multivariable MR analyses were further conducted to ensure the independence of causal association. Reverse-direction MR analyses were performed to rule out the possibility of reverse associations. Finally, enrichment analyses were used to investigate the biofunction. RESULTS: After correction, three microbial taxa were identified to be causally associated with diverse joint OA (PFDR < 0.100), namely Methanobacteriaceae family for knee OA (PFDR = 0.043) and any OA (PFDR = 0.028), Desulfovibrionales order for knee OA (PFDR = 0.045) and Ruminiclostridium5 genus for knee OA (PFDR = 0.063). In addition, we also identified five suggestive microbial taxa that were significant with three different methods under the nominal significance (P < 0.05). Sensitive analysis excluded the influence of heterogeneity and horizontal pleiotropy and multivariable MR analysis ruled out the possibility of horizontal pleiotropy of BMI. GO enrichment analysis illustrates the protective mechanism of the identified taxa against OA. CONCLUSIONS: This study found that several microbial taxa were causally associated with diverse joint OA. The results enhanced our understanding of gut microbiota in the pathology of OA.

Genome-wide identification of m6A-associated SNPs as potential functional variants for bone mineral density.

This study investigated the effect of the N6-methyladenosine (m6A)-associated SNPs on bone mineral density (BMD) and found plenty of m6A-SNPs that were associated with BMD. This study increases our understanding on the regulation patterns of SNP and may provide new clues for further detection of functional mechanism underlying the associations between SNPs and osteoporosis. INTRODUCTION: m6A plays critical roles in many fundamental biological processes and a variety of diseases. The m6A-associated SNPs may be potential functional variants for BMD. The aim of this study was to investigate the effect of the genome-wide m6A-SNPs on BMD. METHODS: We examined the association of m6A-SNPs with femoral neck (FN) and lumbar spine (LS) BMD in 32,961 individuals and quantitative heel ultrasounds (eBMD) in 142,487 individuals. Furthermore, we performed expression quantitative trait locus (eQTL) analyses for the m6A-SNPs using whole genome data of about 10.5 million SNPs and 21,323 mRNAs from 43 Chinese individuals, as well as public available data. Differential expression analyses were also performed to support the identified genes. RESULTS: We found 138, 125, and 993 m6A-SNPs which were associated with FN-BMD, LS-BMD, and eBMD (P < 0.05), respectively. The associations of rs11614913 (P = 8.92 × 10-10) in MIR196A2 and rs1110720 (P = 2.05 × 10-10) in ESPL1 with LS-BMD reached the genome-wide significance level. In addition, a total of 24 m6A-SNPs were significantly associated with eBMD (P < 5.0 × 10-8). Further eQTL analyses showed that 47 of these BMD-associated m6A-SNPs were associated with expressions of the 46 corresponding local genes. Moreover, the expressions of 26 of these genes were associated with BMD. CONCLUSION: The present study represents the first effort of investigating the associations and the mechanisms underlying the link between m6A-SNPs and BMD. The results suggested that m6A-SNP may play important roles in the pathology of osteoporosis.

Association of 3q13.32 variants with hip trochanter and intertrochanter bone mineral density identified by a genome-wide association study.

We performed a GWAS of trochanter and intertrochanter bone mineral density (BMD) in the Framingham Heart Study and replicated in three independent studies. Our results identified one novel locus around the associated variations at chromosomal region 3q13.32 and replicated two loci at chromosomal regions 3p21 and 8q24. Our findings provide useful insights that enhance our understanding of bone development, osteoporosis, and fracture pathogenesis. INTRODUCTION: Hip trochanter (TRO) and intertrochanter (INT) subregions have important clinical relevance to subtrochanteric and intertrochanteric fractures but have rarely been studied by genome-wide association studies (GWASs). METHODS: Aiming to identify genomic loci associated with BMD variation at TRO and INT regions, we performed a GWAS utilizing the Framingham Heart Study (FHS, N = 6,912) as discovery sample and utilized the Women's Health Initiative (WHI) African-American subsample (N = 845), WHI Hispanic subsample (N = 446), and Omaha osteoporosis study (N = 971), for replication. RESULTS: Combining the evidence from both the discovery and the replication samples, we identified one novel locus around the associated variations at chromosomal region 3q13.32 (rs1949542, discovery p = 6.16 × 10-8, replication p = 2.86 × 10-4 for INT-BMD; discovery p = 1.35 × 10-7, replication p = 4.16 × 10-4 for TRO-BMD, closest gene RP11-384F7.1). We also replicated two loci at chromosomal regions 3p21 (rs148725943, discovery p = 6.61 × 10-7, replication p = 5.22 × 10-4 for TRO-BMD, closest gene CTNNB1) and 8q24 (rs7839059, discovery p = 2.28 × 10-7, replication p = 1.55 × 10-3 for TRO-BMD, closest gene TNFRSF11B) that were reported previously. We demonstrated that the effects at both 3q13.32 and 3p21 were specific to the TRO, but not to the femoral neck and spine. In contrast, the effect at 8q24 was common to all the sites. CONCLUSION: Our findings provide useful insights that enhance our understanding of bone development, osteoporosis, and fracture pathogenesis.

Genetic variants in the SOX6 gene are associated with bone mineral density in both Caucasian and Chinese populations.

SUMMARY: Given the biological function of SOX6 and recent genome-wide association finding, we performed a fine-mapping association analyses to investigate the relationship between SOX6 and BMD both in Caucasian and Chinese populations. We identified many single-nucleotide polymorphisms (SNPs) within or near the SOX6 gene to be significantly associated with hip bone mineral density (BMD). INTRODUCTION: SOX6 gene is an essential transcription factor in chondrogenesis and cartilage formation. Recent genome-wide association studies (GWAS) detected a SNP (rs7117858) located at the downstream of SOX6 significantly associated with hip BMD. METHODS: Given the biological function of SOX6 and the GWAS finding, we considered SOX6 as a new candidate for BMD and osteoporosis. Therefore, in this study, we performed a fine-mapping association analyses to investigate the relationship between SNPs within and near the SOX6 gene and BMD at both hip and spine. A total of 301 SNPs were tested in two independent US Caucasian populations (2,286 and 1,000 unrelated subjects, respectively) and a Chinese population (1,627 unrelated Han subjects). RESULTS: We confirmed that the previously reported rs7117858-A was associated with reduced hip BMD, with combined P value of 2.45 × 10(-4). Besides this SNP, we identified another 19 SNPs within or near the SOX6 gene to be significantly associated with hip BMD after false discovery rate adjustment. The most significant SNP was rs1347677 located at the intron 3 (P = 3.15 × 10(-7)). Seven additional SNPs in high linkage disequilibrium with rs1347677 were also significantly associated with hip BMD. SNPs in SOX6 showed significant skeletal site specificity since no SNP was detected to be associated with spine BMD. CONCLUSION: Our study identified many SNPs in the SOX6 gene associated with hip BMD even across different ethnicities, which further highlighted the importance of the SOX6 gene influencing BMD variation and provided more information to the understanding of the genetic architecture of osteoporosis.

The differences of femoral neck geometric parameters: effects of age, gender and race.

UNLABELLED: This study aims at investigating the effects of age, sex, and ethnicity on five femoral neck geometric parameters (FNGPs): femoral neck periosteal diameter, cross-sectional area, cortical thickness, sectional modulus, and buckling ratio and found that the three factors would influence the FNGPs. INTRODUCTION: Bone geometry is one of the most important predictors of bone strength and osteoporotic fractures. This study aims at investigating the effects of age, sex, and ethnicity on five femoral neck geometric parameters (FNGPs): femoral neck periosteal diameter (W), cross-sectional area (CSA), cortical thickness (CT), sectional modulus (Z), and buckling ratio (BR). METHODS: In the studied 861 Caucasian subjects and 3,021 Chinese individuals, CSA, CT, and Z displayed trends of decrease with age, but W and BR showed increasing trends with age in both Chinese and Caucasian females and males (p < 0.05). W, CSA, CT, and Z were significantly higher (p

Searching for genes underlying susceptibility to osteoporotic fracture: current progress and future prospect.

INTRODUCTION: Osteoporotic fracture (OF) is a public health problem. It is a common practice in the genetics of osteoporosis that bone mineral density (BMD) was studied as a major surrogate phenotype in gene search for risk of OF (ROF) because of their high phenotypic correlation. However, some studies indicate that the genetic correlation between BMD and ROF is very low. This implies that most genes found important for BMD may not be relevant to ROF. Ideally, employing OF per se as a direct study phenotype can directly find the relevant genes underlying ROF. EVIDENCE: Here, we summarized some evidence supporting ROF under moderate genetic control, and the current progress of molecular genetic studies employing OF as the direct study phenotype, then give our consideration on the future prospects in the genetics of ROF.

Ethnic difference in osteoporosis-related phenotypes and its potential underlying genetic determination.

Osteoporosis is a serious health problem in both Caucasians and Asians. Caucasians and Asians are two distinct major ethnic groups, which may have differential genetic determination underlying complex genetic diseases such as osteoporosis. However, to date, there has been no systematic review focusing on the aspect of ethnic difference in risk to osteoporosis and its potential underlying genetic determination between Asians and Caucasians. Here, we firstly review diverse aspects of osteoporosis-related differences, including the differences of epidemiology of osteoporotic fractures, peak bone mass, bone loss, bone area, bone geometry and drug treatment response between Asians and Caucasians. Then, we provide some potential genetic evidence on the different heritability and inheritance mode of bone phenotypes, the different osteoporosis candidate genes and the differential results in related molecular studies between them, to explain the above osteoporosis-related phenotypic differences. The results suggest that the osteoporosis-related phenotypic differences between Asians and Caucasians may be partially the result of the different ethnic genetic background. The present review may increase our understanding of potential different mechanisms related to ethnicity in pathogenesis of osteoporosis for effective and potentially customized treatments in different major ethnic groups.

Relationship of total body fatness and five anthropometric indices in Chinese aged 20-40 years: different effects of age and gender.

OBJECTIVES: We aim to evaluate the ethnic-specific relationship of total fat mass and anthropometric indices in Chinese. DESIGN: Cross-section study. SETTING: This study was performed at the College of Life Sciences, Hunan Normal University, P.R. China. SUBJECTS AND METHOD: To increase our understanding of the relationship of total fat mass and anthropometric indices in Chinese, 793 females and 1091 males aged 20-40 years were randomly recruited from Changsha city of P. R. China. Hip circumference (HC) and waist circumference (WC) were measured using standardized equipments, and other three anthropometric indices of body mass index (BMI), waist-to-hip ratio (WHR), and conicity index (CI) were calculated using weight, height, HC and WC. Total body fatness (TBF) in kg was measured using a Hologic QDR 4500 W dual-energy X-ray absorptiometry (DEXA) scanner. RESULTS: There was an increasing trend of TBF, %TBF (percent total body fatness) and the five anthropometric indices in successively older age groups. Compared with females, males generally had high average BMI, WC, HC, WHR and CI, but had low average TBF and %TBF. Except for some correlations in 25-29 years age groups, TBF and %TBF were significantly correlated with five anthropometric indices with the Pearson's correlation coefficients ranging from 0.07 to 0.87. Principal component analysis (PCA) was performed to form four principal components (PCs) that interpreted over 99% of the total variation of the five related anthropometric indices in all age groups, with over 53% of the total variation accounted for by the PC1. Multiple regression analyses showed that four PCs combined explained a greater variance (R (2)=55.2-80.8%) in TBF than did BMI alone (R (2)=40-74.9%). CONCLUSION: Our results suggest that there is an increasing trend of total fat mass and five anthropometric indices with aging; that age and sex have the important effects on influencing the correlations of TBF and the studied anthropometric indices; and that the accuracy of predicting the TBF using five anthropometric indices is higher than using BMI alone.

Tests of linkage and association of the COL1A2 gene with bone phenotypes' variation in Chinese nuclear families.

In the present study, we simultaneously test linkage and/or association of the collagen type I alpha 2 (COL1A2) gene with bone mineral density (BMD) and bone area. A total of 1280 subjects from 407 Chinese nuclear families (including both parents and their daughters) were genotyped for an intragenic marker MspI in the COL1A2 gene. BMD and bone area at the lumbar spine and hip were measured by dual-energy X-ray absorptiometry. Applying the QTDT (quantitative transmission disequilibrium test) program, we performed tests for population stratification, within-family association (via transmission disequilibrium test), total association, linkage, and linkage while modeling association. Significant or marginal within-family associations were found with BMD at the lumbar spine (P = 0.013), trochanter (P = 0.004), and total hip (P = 0.053) and with bone area at the intertrochanteric region (P = 0.024) and total hip (P = 0.048). The positive associations were confirmed in permutations except for bone area at total hip (P > 0.10). A small proportion (<1%) of the population variance of bone phenotypes can be explained by the MspI polymorphism; however, it may be underestimated given the significant population stratification detected in our sample. Due to the limited number of sib pairs in this sample, we did not find evidence of linkage. In summary, the MspI polymorphism is likely to be in linkage disequilibrium with a nearby functional mutation affecting BMD and bone area.

Differentiation of Caucasians and Chinese at bone mass candidate genes: implication for ethnic difference of bone mass.

Bone mineral density (BMD) is an important risk factor for osteoporosis and has strong genetic determination. While average BMD differs among major ethnic groups, several important candidate genes have been shown to underlie BMD variation within populations of the same ethnicity. To investigate whether important candidate genes may contribute to ethnic differences in BMD, we studied the degree of genetic differentiation among several important candidate genes between two major ethnic groups: Caucasians and Chinese. The genetic variability of these two populations (1131 randomly selected individuals) was studied at six restriction sites exhibiting polymorphisms of five important candidate genes for BMD: the BsaHI polymorphism of the calcium-sensing receptor (CASR) gene, the SacI polymorphism of the alpha2HS-glycoprotein (AHSG) gene, the PvuII and XbaI polymorphisms of the estrogen receptor alpha (ESR1) gene, the ApaI polymorphism of the vitamin D receptor (VDR) gene, and the BstBI polymorphism of the parathyroid hormone (PTH) gene. The two ethnic groups showed significant allelic and genotypic differentiation of all the polymorphisms studied. The mean FST was 0.103, which significantly differed from zero (P < 0.01). The Chinese population had lower mean heterozygosity (0.331) than the Caucasian one (0.444); the CASR-BsaHI and PTH-BstBI polymorphisms contributed most significantly to this difference. Analysis of the intra- and inter-population variability suggests that various types of natural selection may affect the observed patterns of variation at some loci. If some of the candidate genes we studied indeed underlie variation in BMD, their population differentiation revealed here between ethnic groups may contribute to understanding ethnic difference in BMD.

The effects of reverberant blast waves on the auditory system.

Chinchillas were exposed to 1, 10, or 100 reverberant impulses at 150, 155, or 160 dB peak SPL. The impulses were generated by one of two different shock tubes, each producing blast waves having a different spectral composition, with one emphasizing low frequencies (< 0.5 kHz) and the other midfrequencies (2-4 kHz). Impulses were presented at the rate of one per minute. This parametric paradigm yielded 18 exposure conditions with 15 animals/condition. Hearing thresholds were measured using auditory-evoked potentials and the sensory epithelium was evaluated with the surface preparation. In general, trauma increased as the total energy of the exposure, determined by the peak SPL and number of presentations, increased. The dependent variables (permanent threshold shift and sensory cell loss) varied in an orderly fashion across frequency as the peak and number of presentations were increased for both blast wave sources. There were, however, consistent differences between the effects of the low- and high-frequency energy "content" blast waves. Correlations between the dependent variables and the energy of exposure were highest for P- or A-weighted energies [Patterson et al., J. Acoust. Soc. Am. 93, 2860-2869 (1993)].

The cubic distortion product otoacoustic emissions from the normal and noise-damaged chinchilla cochlea.

A normative study of the cubic distortion product emissions from 104 monaural and binaural chinchillas was undertaken to establish criteria upon which noise exposed animals could be evaluated. From this normative group, 47 randomly selected chinchillas were exposed to various high level (150-, 155-, and 160-db peak SPL) impulse noises. Auditory evoked potentials and cubic distortion product otoacoustic emissions were measured on each animal pre- and post-exposure and related to the sensory cell populations 30 days post-exposure. Both group mean and individual animal data indicated that the distortion product emissions were more sensitive, frequency-specific indices of noise-induced cochlear effects than pure-tone threshold measures. This was particularly evident near the threshold for noise-induced damage to the outer hair cell system.

The application of frequency and time domain kurtosis to the assessment of hazardous noise exposures.

Five computer-synthesized broadband noises, each having the same average spectrum and the same unweighted Leq of 100 dB SPL but very different temporal structures, were used to produce hearing loss in chinchillas. Despite the same exposure energies and spectra, each noise exposure produced a different magnitude and frequency distribution of hearing loss and sensory cell loss. The results indicate that the statistical properties of a signal are important in the determination of hearing loss. When the audiometric and histological results are compared to a metric based upon kurtosis measured in the time and the frequency domain for each exposure, there is a clear indication that these statistical metrics are good predictors of the relative magnitude and frequency distribution of the acoustic trauma.

Hearing threshold shifts from repeated 6-h daily exposure to impact noise.

Exposure of chinchillas to broadband, high-level impact noise on an interrupted 6-h daily schedule over 20 days has shown that pure-tone thresholds measured immediately following each daily exposure improve as much as 30 dB despite the continuing noise exposure. The time constant of this recovery effect (toughening) and the magnitude of the effect are related to the audiometric test frequency and the exposure energy. The trauma, quantified by permanent threshold shifts and sensory cell losses, produced by the interrupted exposure paradigm is generally less than that produced by an equal-energy uninterrupted exposure. The wide variations in the temporal pattern of threshold shift across similarly exposed animals suggest that the toughening effect reflects the underlying susceptibility of that animal to noise trauma.

Audiometric and histological differences between the effects of continuous and impulsive noise exposures.

An experiment was designed to determine if, for equal SPL and power spectrum, the effects on hearing of high-kurtosis noise exposures and a Gaussian noise exposure are different and the extent to which any differences measured in terms of audiometric and histological variables are frequency specific. Three groups of chinchillas with 10 animals/group were exposed for 5 days at 90 dB SPL to one of three types of noise, each with the same power spectrum. The impulsiveness, defined by the kurtosis, and the region of the spectrum from which the impulsive components of the noise were created differed for two of the noises, while the third was a continuous Gaussian noise. The results show that the most impulsive noise produced up to 20 dB greater permanent threshold shift at the high frequencies than did the Gaussian noise exposure. However, these audiometric results were difficult to reconcile with the pattern of sensory cell losses that showed statistically significant larger losses of outer hair cells for the impulsive exposure in the 0.25-kHz region. When the impacts in a high-kurtosis noise were created from the energy in the 1- through 6-kHz region of the spectrum, the audiometric profile of hearing loss was similar to that produced by the Gaussian noise; however, inner hair cell losses were significantly greater in the 4-kHz octave band region of the cochlea.