Search

Objective. The aim of the investigation was to study the possible association of the rs7574865 polymorphism of the STAT4 gene with syndesmophytes (SMP) of the spine in patients with ankylosing spondylitis (AS).Subjects and methods. The study included a cohort of 100 patients, 79 men and 21 women with a diagnosis of AS.All patients were positive for the HLA-B27 antigen, had a mean age of 39.6±10.9 years and a mean disease duration of 60,4±28,4 months. The association of the rs7574865 polymorphism of the STAT4 gene with the SMP of the cervical, thoracic and lumbar spine was studied. For genotyping of the rs7574865 polymorphism, the method of allele-specific polymerase chain reaction in real time (RT-PCR) was used.Results. Spearman’s correlation analysis showed a statistically significant positive relationship between SMP in the thoracic spine and rs7574865 polymorphism of the STAT4 gene (r=0.23; p=0.022). The frequency of GT genotype carriers in the group of patients with thoracic spine trSMP(+) was statistically significantly lower than in the alternative group trSMP(–) (28.2% and 50.8%, respectively; p=0.025). Carriage of the GT genotype in patients with AS reduced the risk of trSMP(+) formation in the thoracic spine (OR=0.31) and this genotype was protective. No reliably significant association of the studied polymorphism with SMP of the cervical and lumbar spine was found. Patients with trSMP(+) were statistically significantly older in age, had a longer duration of the disease and a higher functional BASFI index compared with patients without trSMP(–).Conclusion. Genetic testing of the rs7574865 G/T polymorphism of the STAT4 gene in patients with AS opens up the possibility of using this polymorphism as a genetic marker-predictor – X-ray progression of structural changes in the thoracic spine.

The aim of the study was investigation of association of the rs7574865 polymorphism of STAT4 gene (Signal Transducer and Activator of Transcription 4, a family of signal transduction and transcription activation molecules 4) with very early rheumatoid arthritis (RA) in Russian patients, and the study of the relationship of "phenotype – genotype", particularly of positivity for antibodies to cyclic citrullinated peptides (ACCP), their concentration, the presence of erosive joint damage at inclusion in patients the study with the STAT4 gene polymorphism. Material and methods. The study was conducted in the framework of the program REMARKA (Russian study of Methotrexate and biological drugs in Early active Arthritis). 85 patients with very early RA with a duration of symptoms no more than 6 months, not receiving disease modifying anti-rheumatic drugs (DMARD) and biologicals were included. Results and discussion. The analysis of the distribution of genotypes and alleles of STAT4 rs7574865 polymorphism showed that the frequencies of g/G, G/T and T/T genotypes differ at the level of the prominent tendency to statistical significance between patients with RA and the control group (p=0.05). The frequency of minor allele T in RA is significantly higher than that in the control group, and this allele is associated with a predisposition to RA [odds ratio (OR) 1.7; 95% confidence interval (CI) 1.1–2.8; p=0.03]. In the study of the "genotype – phenotype" relationship, STAT4 gene polymorphism correlated with erosive joint damage (r2=0.289; p=0.008). In carriers of the homozygous genotype TT, the number of erosions at inclusion in the study was significantly higher compared with carriers of genotypes GG/GT (median 5.50 [0.754; 7.5] and 0.00 [0.00; 2.00], respectively; p=0.003). The risk of erosion is also associated with the polymorphism of the STAT4 gene (OR 8.6; 95% CI 1.0–204.9; p=0.03). A difference of the ACCP level depending on STAT4 gene polymorphism was revealed: carriers of at least one minor allele T (G/T+T/T) had significantly higher concentration of ACCP than that in carriers of homozygous GG genotype: 248.97±151.00 and 179.51±147.01 U/ml, respectively; p=0.048).Conclusion. The study showed that in Russian patients with very early RA, the STAT4 gene rs7574865 polymorphism is associated with both predisposition to the disease and prognostically unfavorable manifestations (phenotypes) of the disease, namely, with the development of erosion in the early stages of the disease and an increase of the ACCP level.

Objective: to investigate the impact of VDR and MCP-1 gene polymorphisms on the efficiency of 12-month therapy with strontium ranelate (SR) in women with postmenopausal osteoporosis (OP). Subjects and methods. The investigation enrolled 34 postmenopausal women (mean age 65±8 years), who was diagnosed with OP by lumbar spine and/or proximal femur examinations using a Hologic QDR 4500W X-ray bone densitometer. The gene polymorphisms of VDR (BsmI) and MCP-1 (-2518A/G) were genotyped by polymerase chain reaction, followed by restriction fragment length polymorphism analysis. Results and discussion. The carriers of the BB and Вb genotypes of the VDR gene had a statistically significantly lower increase in the bone mineral density (BMD) of the lumbar spine than those of the homozygous recessive bb genotype (p = 0.024 and p = 0.022, respectively) and the bb genotype and the BB+Bb genotype (p = 0.004), and the carriers of the GА genotype of MCP-1 gene had a statistically significantly lower increase in the BMD of the entire proximal femur than those of the homozygous AA genotype (0.2±2.5% and 4.4±4.4%, respectively; p = 0.004) at 12 months after SR treatment. Conclusion. The findings may suggest that the BB and Bb genotypes of the VDR gene and the GA genotype of the MCP-1 gene may negatively affect the efficacy of SR in patients with OP. However, additional studies on a larger sample of patients, including those with other forms of OP, are required to confirm this assumption.

Numerous clinical observations show that the human genetic background plays an important role in predisposition to many diseases. Objective: to investigate a possible relationship of the polymorphisms in 19 A/G leptin (LEP) gene, in the interleukin (IL)-1 receptor antagonist (IL-1RA) gene VNTR (variable number of tandem repeats), and in the -174G/C IL-6 gene to the risk of developing the clinical phenotypes of panniculitis (PN), clinical and laboratory parameters. Subjects and methods. The study enrolled 54 patients (48 women and 6 men) aged 15 to 76 years with a documented diagnosis of PN who had been treated at the V.A. Nasonova Research Institute of Rheumatology. Group 1 of 46 patients with erythema nodosum (EN) and Group 2 of 8 with Weber-Christian panniculitis (WCP) were formed for genetic study. The genotyping data on 197 healthy unrelated individuals were used as a control. Clinical and laboratory information was available from 39 patients for genotyping 19A/G LEP gene polymorphism, that from 43 patients for -174G/C IL-6 gene polymorphism, and that from 46 patients for IL-1RA polymorphism. Genotyping was performed by a polymerase chain reaction-restriction fragment length polymorphism (PCRRFLP) analysis. Results and discussion. The frequencies of the LEP 19 GG genotype and the LEP G allele in patients with EN and WCP were significantly higher than those in the controls (48.7 and 18.2%, p = 0.0004; 50.0 and 18.2%, p = 0.053; and 70.5 and 45.4%, p = 0.0002, respectively). The frequencies of the A1A1 genotype and A1 allele of IL-1RA gene VNTR polymorphism in EN group were significantly higher than those in the controls (67.4 and 44.2%, p=0.011; 80.4 and 61.6%, p = 0.002, respectively). The frequency of IL-6 -174GC polymorphism was also higher in the EN group than that in the controls (58.1 and 34.7%, p = 0.008). The -174G/C polymorphism showed a significant association with the site of erythema (p = 0.028). In patients with a single lesion of erythema on the body, the frequency of the -174 GC genotype was significantly higher than that in those with multiple foci of erythema (72.0 and 31.2%, respectively; p = 0.025). In the WCP group, analysis of variance showed an association of IL-1RA gene VNTR polymorphism with the intensity of pain, assessed by the visual analogue scale. The carriers of the A1A1 genotype had more severe pain than those of the A1A2 genotype (83.3±11.5 and 20.0±18.2 mm, respectively; p = 0.008). Conclusion. The findings suggest that genetic testing can be used to predict the clinical course of PN.

Genetic factors that are an important hereditary component determining a predisposition to osteoporosis (OP) are 60–80% responsible for bone mineral density (BMD). Some polymorphic genes have been previously shown to affect the efficiency of performed anti-osteoporotic therapy. Objective: to study the impact of farnesyl diphosphate synthase (FDRS) and geranylgeranyl diphosphate synthase (GGSPI) gene polymorphisms on BMD changes during 12-month therapy with bisphosphonates (BP) in women with postmenopausal OP. Subjects and methods. The investigation enrolled 53 women with OP. Spine and proximal femur BMD was determined using X-ray densitometry before and after BP treatment. The -99A/C and -8188T ins/del polymorphisms in the FDPS and GGPS1 genes were investigated using real-time polymerase chain reaction. Results and discussion. The BMD changes were less marked in women with the C allele of C/T -99/C polymorphism in the FDPS gene than those in carriers of the genotype AA: 2.3±3.6 and 4.4±3.8% (р = 0.062) in the spine; 0.6±3.1 and 2.8±4.5% (р = 0.075) in the femoral neck; 0.5±2.9 and 2.5±2.8% (р = 0.020) in the entire femur, respectively. Femoral neck densitometry showed a significantly weaker response to BP treatment in the patients carrying the mutant genotype del/del of GGSP1 -8188T ins/del polymorphism than in those with the wild-type genotype ins/ins (0.8±4.2 and 4.1±2.5%, respectively; р = 0.030). No significant differences for this polymorphism were found in other areas of BMD measurement. Conclusion. The described pilot study has indicated that the examined FDPS and GGSP1 gene polymorphisms may be predictors for a response to BP therapy in patients with OP. Further investigations that will contribute to the choice of the most effective therapy for this disease are needed to confirm our results.

Many clinical observations show that patient’s genetic background is of great importance in determining the efficiency of treatment. Subjects and methods. The instigation included 50 postmenopausal women with osteoporosis (OP), who were followed up at the Laboratory of osteoporosis, V.A. Nasonova Research Institute of Rheumatology. Body mineral density (BMD) in the lumbar spine (LI-IV), femoral neck (FN), and total hip was measured using dual-energy X-ray absorptiometry before and 12 months after treatment with bisphosphonates (BP). To estimate BMD changes, the investigators used ΔBMD in percent (Δ, %). Results and discussion. The whole group showed a positive effect of BP therapy during a year, which was most pronounced in the lumbar spine (mean ΔBMD, about 4%), and a small increment in the proximal hip BMD (mean ΔBMD, about 2%). An analysis indicated a statistically significant correlation of MCP1 -2518A>G polymorphism with changes in LI-IV BMD after 12-month BP therapy. Thus, the female patients who were A allele carriers had a twice lower increase in LI-IV BMD due to BP therapy than those without this allele. The genetic variants of the CCR5 gene, which were related to Δ32 deletion, and IL1β -511C/T polymorphism were also associated with changes in FN BMD following 12-month BP therapy. The BMD increase due to BP therapy in the carriers of the CCR5 Δ32 mutation (wt/Δ32 genotype) was 3.5-fold than that in the carriers of the wild type gene (wt/wt genotype). Examination of IL1 -511C/T polymorphism demonstrated that the FN BMD increment in the carriers of the CC genotype was significantly higher than in those of the CT genotype (4.2±4.8 and 1.0±3.7%, respectively; р = 0.023). Our investigation revealed no significant relationship between VDR, LEPR, IL10, MHTFR, PPARG, SPP1, and CCR5(G/A) gene polymorphisms and 12-month BP therapy-induced BMD changes in the three study skeletal regions. The findings may suggest that genetic testing may be used to predict a response to BP.

Interleukin-1β (IL-1β) is a potential stimulant of bone resorption. IL-1β receptor antagonist (IL-1RA) is a natural inhibitor of the biological effects of IL-1β. Objective: to study the frequency distribution of polymorphisms in the IL-1β and IL-1RA genes and their association with bone mineral density (BMD) in women with primary osteoporosis (OP). Subjects and methods. The distribution of genotype frequency of IL-1β (-511C/T) polymorphism and that of IL-1RA 511C/T polymorphism that is associated with the number of variable tandem repeats (VTR), were investigated in 254 women with OP and 214 healthy women. Results and discussion. IL-1β (-511C/T) genotype carriers were encountered somewhat more frequently among the patients with OP (53.0%) than in the control group (43.4%); however, the differences were insignificant. In these carriers, the risk of OP was 1.5-fold higher than that in those of other genotypes (odds ratio, 1.49; confidence interval, 1.02–2.18; p = 0.041). The patients who were IL-1β T allele (CT- and TT-genotype) carriers had a significantly lower spine (LI–IV) BMD than those who had not this allele (CC-genotype; p = 0.011). The patients and the controls showed no differences in the frequency distribution of IL-1RA gene polymorphism associated with the number of VTR. In the OP group, the carriers of the rare genotype A1A3 in the IL-1RA gene (3.1%) had significantly higher femoral neck BMD (0.698±0.064 g/cm2) than those of the А1А1, А1А2 and А2А2 genotypes (0.613±0.078; 0.607±0.082. and 0.615±0.064 g/cm2; р = 0.003, р = 0.003, and р = 0.002, respectively). Conclusion. IL-1β (-511C/T) polymorphism is associated with lower spine BMD and IL-1RA A1A3 genotype polymorphism is related to higher femoral neck BMD.

Objective: to study an association of bone mineral density (BMD) with the expression of genes associated with the proliferation, survival, and differentiation of osteocytes in the peripheral blood of postmenopausal women with osteoporosis (OP). Subjects and methods. Twenty-eight postmenopausal women with OP and 17 age-matched healthy women were examined. BMD was measured by X-ray absorptionmetry. Alkaline phosphatase activity and calcium and phosphorus levels in the blood were determined by conventional methods. RNA was isolated from the peripheral blood and used to estimate the gene expression, by using real-time polymerase chain reaction (RT-PCR). Results. The postmenopausal women with OP were observed to have a significant increase in the expression of ATG1 (serine-threonine kinase that is responsible for the generation of autophagic vacuoles) as compared to the healthy controls, which suggests that autophagy develops in the blood cells of these patients. This was attended by a considerable reduction in the expression of the mTOR (the mammalian target of rapamycin) gene, a regulator of protein synthesis and cell proliferation, and in that of the genes associated with the differentiation of osteoblasts: transforming growth factor β 1 (TGF- β 1), Runx 2 (Runt-related transcription factor 2) and total alkaline phosphatase (TAP), and the proinflammatory cytokines tumor necrosis factor- α (TNF- α ) and interleukin 1 β (IL-1 β ). The expression of mTOR, TGF- β 1, Runx2, and IL-1 β genes was positively correlated with BMD in individual areas of the hip. On the contrary, that of ATG1 and TAP was correlated negatively with BMD in this area and positively with that in the spine. Although the expression of the genes in question was not correlated with the serum levels of phosphorus and calcium, that of TGF- β 1 was positively related to TAP activity in postmenopausal women with OP. Conclusion. In postmenopausal women with OP, bone loss is accompanied by a significant increase in the expression of the autophagy marker ATG1 and by a reduction in that of mTOR, which may be indicative of the inhibition of protein biosynthesis and cell proliferation and the suppression of the expression of the genes associated with the differentiation of osteoblasts and the proinflammatory cytokines TNF- α and IL-1 β . The obtained data on gene expression may be used to search for new disease markers tested in peripheral blood.

Polymorphisms of 21 genes involved in the processes of bone remodeling were studied on samples of 150 to 265 postmenopausal healthy women in a control group and those of 175 to 269 patients with osteoporosis (OP). In a Moscow sample of postmenopausal women, the genotypes of OP susceptibility were found to be the CT genotype of the low-density lipoprotein receptor-related protein 5 (LRP5) gene, the GG genotype of the leptin (LEP) gene, the XX genotype of the estrogen receptor a (ER-a) gene, the Ff genotype of the vitamin D receptor (VDR) gene, the HH genotype of the a1 polypeptide chain of type II collagen gene and the GG genotype of the aromatase (CYP19) gene. The interaction of a number of genes to develop OP susceptibility was ascertained to be compliant. For example, the carriage of the CTxx genotypes of the LRP5/ERa genes, the CTGG genotypes of the LRP5/CBFA genes, the CCGA genotypes of the TGF/31/CYP19 genes, and the CCAA genotypes of the TGF/31/OPG genes increases a risk for OP by 7.7, 4.1, 6.2, and 2.7 times, respectively. The genotypes increasing the risk of spinal osteoporotic fractures were AG of 163A/G polymorphism in the osteoprotegerin (OPG) gene, TT of 509C^T polymorphism in the TGF/31 gene, and BB of BsmI polymorphism in the VDR gene (OR = 4.9, 3.9, and 4.4, respectively). The genotype of a risk for osteoporotic fractures at other sites was AG of A19G polymorphism in the LEP gene (OR = 2.6).

Objective: to study the mechanism for the involvement of ТGFβ1 T(861-20)C in bone resorption in postmenopausal osteoporosis (OP). Material and methods. DNA from 158 postmenopausal women and patients with OP and from 89 healthy age-matched women was examined by polymerase chain reaction (PCR)-restriction fragment length polymorphism (PCR-RFLP) analysis. Bone mineral density (BMD) was estimated by dual-energy X-ray absorptiometry. Standard biochemical protocols were used to detect alkaline phosphatase activity and calcium and phosphorus levels in serum. Total RNA was isolated from the peripheral blood of 32 patients with OP and 39 healthy donors and used for real-time PCR study. Results. No significant differences were found in the frequency of individual alleles and genotypes between the OP group and control donors. The minor T allele frequency was 0.27. There was a significant correlation of ТGFβ1 T(861-20)C polymorphism with low lumbar spine BMD (r=0.18; p=0.025) in Russian patients with OP. Age-adjusted (Z-score) BMD in CC genotype carriers turned to be significantly lower than that in CT and TT genotype carriers. This was accompanied by lower ТGFβ1 gene expression in the peripheral blood of CC genotype carriers (n=10) as compared to the combined group of carriers of two other genotypes (n=22) in the OP group (p=0.03). No changes in ТGFβ1 gene expression were seen in healthy women who were CC genotype carriers (n=18) as compared to the combined representatives of two other genotypes (n=21). Overall, the OP group exhibited significantly lower ТGFβ1 gene expression than the healthy controls (p=0.04). Conclusion. The association of ТGFβ1 (861-20)CC genotype with lower lumbar spine BMD in patients with OP is attended by decreased ТGFβ1 gene expression. Therefore, ТGFβ1 T(861-20)C polymorphism may be a predictor for the development of OP and the more severe form of the disease may be expected in (861-20)CC genotype carriers.