Toshiaki Kotani

A genome-wide association study identifies common variants near LBX1 associated with adolescent idiopathic scoliosis

Adolescent idiopathic scoliosis is a pediatric spinal deformity affecting 2–3% of school-age children worldwide. Genetic factors have been implicated in its etiology. Through a genome-wide association study (GWAS) and replication study involving a total of 1,376 Japanese females with adolescent idiopathic scoliosis and 11,297 female controls, we identified a locus at chromosome 10q24.31 associated with adolescent idiopathic scoliosis susceptibility. The most significant SNP (rs11190870; combined P = 1.24 × 10−19; odds ratio (OR) = 1.56) is located near LBX1 (encoding ladybird homeobox 1). The identification of this susceptibility locus provides new insights into the pathogenesis of adolescent idiopathic scoliosis.

Genetic variants in GPR126 are associated with adolescent idiopathic scoliosis

Adolescent idiopathic scoliosis (AIS) is the most common pediatric skeletal disease. We previously reported a locus on chromosome 10q24.31 associated with AIS susceptibility in Japanese using a genome-wide association study (GWAS) consisting of 1,033 cases and 1,473 controls. To identify additional AIS-associated loci, we expanded the study by adding X-chromosome SNPs in the GWAS and increasing the size of the replication cohorts. Through a stepwise association study including 1,819 cases and 25,939 controls, we identified a new susceptibility locus on chromosome 6q24.1 in Japanese (P = 2.25 × 10−10; odds ratio (OR) = 1.28). The most significantly associated SNP, rs6570507, was in GPR126 (encoding G protein–coupled receptor 126). Its association was replicated in Han Chinese and European-ancestry populations (combined P = 1.27 × 10−14; OR = 1.27). GPR126 was highly expressed in cartilage, and the knockdown of gpr126 in zebrafish caused delayed ossification of the developing spine. Our results should provide insights into the etiology and pathogenesis of AIS.

Lack of association between adolescent idiopathic scoliosis and previously reported single nucleotide polymorphisms in MATN1, MTNR1B, TPH1, and IGF1 in a Japanese population.

Adolescent idiopathic scoliosis (AIS) is a spinal deformity most commonly arising in apparently healthy girls around puberty. AIS has a strong genetic predisposition. Several genetic associations between AIS and single nucleotide polymorphisms (SNPs) have been reported; common SNPs in the genes for matrilin 1 (MATN1), melatonin receptor 1B (MTNR1B), tryptophan hydroxylase 1 (TPH1), and insulin‐like growth factor 1 (IGF1) are reported to be associated with AIS in Chinese. However, these associations have not been replicated so far. To confirm the associations, we compared these SNPs with AIS predisposition and curve severity in a population of Japanese females consisting of 798 AIS patients and 1,239 controls. All the subjects were genotyped using the PCR‐based Invader assay. We found no association of any of the SNPs with AIS predisposition or curve severity. Considering the statistical power and sample size of the present study, we concluded that these SNPs are not associated with either AIS predisposition or curve severity in Japanese.

Replication study of the association between adolescent idiopathic scoliosis and two estrogen receptor genes.

Adolescent idiopathic scoliosis (AIS) is a common disorder with a strong genetic predisposition. Associations between AIS and common single nucleotide polymorphisms (SNPs) in estrogen receptor genes have been reported. rs9340799 in the gene for estrogen receptor α (ESR1) is reported to be associated with curve severity in Japanese and with AIS predisposition and curve severity in Chinese. In addition, rs1256120 in the gene for estrogen receptor β (ESR2) is reported to be associated with AIS predisposition and curve severity in Chinese. However, the sample sizes of these previous studies were small, and the associations of these SNPs have not been replicated. To examine the association between AIS and estrogen receptor genes, we investigated the association of rs9340799 and rs1256120 with AIS predisposition and curve severity using a large Japanese population, consisting of 798 AIS patients and 637 sex‐matched controls. We found no association of either SNP with AIS predisposition or curve severity in the Japanese population. Considering the statistical power of the present study and the limitations of the previous reports, we conclude that the associations of rs9340799 and rs1256120 with AIS predisposition and curve severity are negative.

Risk factors for complications associated with growing-rod surgery for early-onset scoliosis

Study Design. A retrospective multicenter study. Objective. To identify risk factors for postoperative complications associated with growing-rod (GR) surgery for early-onset scoliosis (EOS). Summary of Background Data. Results and complications of GR surgery for EOS have not been adequately studied. Methods. We evaluated clinical and radiographical results from 88 patients with EOS who underwent GR surgery in 12 spine centers in Japan. The mean age at the time of initial surgery was 6.5 ± 2.2 years (range, 1.5–9.8 yr) and the mean follow-up period was 3.9 ± 2.6 years (range, 2.0–12.0 yr). Risk factors for postoperative complications were analyzed using binomial multiple logistic regression analysis. We considered the potential factors of sex, age, number of rod-lengthening procedures, whether a pedicle screw foundation was used, the uppermost level of the proximal foundation and lowermost level of the distal foundation, Cobb angles of the proximal thoracic, main thoracic, and lumbar curves, and the kyphosis angles in the proximal, main thoracic, thoracolumbar, and lumbar spine. Kaplan-Meier analysis was used to determine the complication-free survival rate of GR surgery as a function of the number of surgical procedures. Results. Complications affected 50 of the patients (57%) and were associated with 119 of 538 surgical procedures, with 86 implant-related failures (72%), 19 infections (16%), 3 neurological impairments (3%), and 11 other complications. The most frequent implant-related failure was dislodged implant (71%) and 95% of the dislodgements occurred at the proximal foundation. Kaplan-Meier analysis demonstrated a linear decrease in complication-free rates as the number of rod-lengthening procedures increased. Binomial multiple logistic regression analysis found the following significant independent risk factors: 6 or more rod-lengthening procedures (odds ratio [OR], 6.534), an increase of every 20° in the proximal thoracic Cobb angle (OR, 3.091), and an increase of every 25° in the lumbar lordosis angle (OR, 2.607) in the preoperative condition. Conclusion. Increases in the upper thoracic scoliotic curve, thoracic kyphosis, and number of rod-lengthening procedures are positively associated with an increased risk of complications after GR surgery for EOS. Level of Evidence: 4

A Functional SNP in BNC2 Is Associated with Adolescent Idiopathic Scoliosis

Adolescent idiopathic scoliosis (AIS) is the most common spinal deformity. We previously conducted a genome-wide association study (GWAS) and detected two loci associated with AIS. To identify additional loci, we extended our GWAS by increasing the number of cohorts (2,109 affected subjects and 11,140 control subjects in total) and conducting a whole-genome imputation. Through the extended GWAS and replication studies using independent Japanese and Chinese populations, we identified a susceptibility locus on chromosome 9p22.2 (p = 2.46 × 10(-13); odds ratio = 1.21). The most significantly associated SNPs were in intron 3 of BNC2, which encodes a zinc finger transcription factor, basonuclin-2. Expression quantitative trait loci data suggested that the associated SNPs have the potential to regulate the BNC2 transcriptional activity and that the susceptibility alleles increase BNC2 expression. We identified a functional SNP, rs10738445 in BNC2, whose susceptibility allele showed both higher binding to a transcription factor, YY1 (yin and yang 1), and higher BNC2 enhancer activity than the non-susceptibility allele. BNC2 overexpression produced body curvature in developing zebrafish in a gene-dosage-dependent manner. Our results suggest that increased BNC2 expression is implicated in the etiology of AIS.

Identification of a susceptibility locus for severe adolescent idiopathic scoliosis on chromosome 17q24.3.

Adolescent idiopathic scoliosis (AIS) is the most common spinal deformity, affecting around 2% of adolescents worldwide. Genetic factors play an important role in its etiology. Using a genome-wide association study (GWAS), we recently identified novel AIS susceptibility loci on chromosomes 10q24.31 and 6q24.1. To identify more AIS susceptibility loci relating to its severity and progression, we performed GWAS by limiting the case subjects to those with severe AIS. Through a two-stage association study using a total of ∼12,000 Japanese subjects, we identified a common variant, rs12946942 that showed a significant association with severe AIS in the recessive model (P = 4.00×10−8, odds ratio [OR] = 2.05). Its association was replicated in a Chinese population (combined P = 6.43×10−12, OR = 2.21). rs12946942 is on chromosome 17q24.3 near the genes SOX9 and KCNJ2, which when mutated cause scoliosis phenotypes. Our findings will offer new insight into the etiology and progression of AIS.

Long-term clinical outcomes of surgery for adolescent idiopathic scoliosis 21 to 41 years later.

Study Design. A case control study. Objective. To determine the clinical outcome of middle-aged patients surgically treated for adolescent idiopathic scoliosis and to compare their outcomes with assessments of age- and sex-matched healthy controls. Summary of Background Data. Several long-term follow-up studies have been published on the clinical outcomes of surgical treatment for adolescent idiopathic scoliosis in patients who have reached their 20s or 30s. However, clinical outcomes in patients who have reached middle age remain unknown. Methods. This study included 256 patients surgically treated for adolescent idiopathic scoliosis (AIS) between 1968 and 1988. The Scoliosis Research Society Patient Questionnaire (SRS-22) and Roland-Morris Disability Questionnaire (RDQ) were used for evaluating long-term clinical outcomes. Sixty-six (25.8%; 62 females, 4 males; mean age, 46.0 years [range 34–56]) of the 256 patients responded to the questionnaires. The mean follow-up period was 31.5 (range 21–41) years. Seventy-six healthy age- and sex-matched individuals with neither a history of spinal surgery nor scoliosis were selected as a control (CTR) group. Results. On the basis of the SRS-22 responses, AIS patients had significantly decreased function (AIS: 4.3 ± 0.6, CTR: 4.7 ± 0.5, P < 0.01) and decreased self-image (AIS: 3.0 ± 0.8, CTR: 3.7 ± 0.5, P < 0.01) in comparison with the controls, but the 2 groups were similar with respect to pain (AIS: 4.3 ± 0.6, CTR: 4.2 ± 0.5, P = 0.14) and mental health (AIS: 3.9 ± 0.9, CTR: 3.7 ± 0.7, P = 0.14). The RDQ responses indicated that low back pain was not significantly increased in the AIS group compared with the CTR group (AIS: 1.8 ± 3.5, CTR: 1.4 ± 3.1, P = 0.36). Conclusion. Surgery had no demonstrable adverse effects on pain or mental health in these middle-aged AIS patients 21–41 years after surgery, although the AIS patients did have significantly lower function and lower self-image than the controls.

Accuracy of Pedicle Screw Placement in Scoliosis Surgery: A Comparison between Conventional Computed Tomography-Based and O-Arm-Based Navigation Techniques.

Study Design Retrospective study. Purpose We compared the accuracy of O-arm-based navigation with computed tomography (CT)-based navigation in scoliotic surgery. Overview of Literature No previous reports comparing the results of O-arm-based navigation with conventional CT-based navigation in scoliotic surgery have been published. Methods A total of 222 pedicle screws were implanted in 29 patients using CT-based navigation (group C) and 416 screws were implanted in 32 patients using O-arm-based navigation (group O). Postoperative CT was performed to assess the screw accuracy, using the established Neo classification (grade 0: no perforation, grade 1: perforation <2 mm, grade 2: perforation ≥2 and <4, and grade 3: perforation ≥4 mm). Results In group C, 188 (84.7%) of the 222 pedicle screw placements were categorized as grade 0, 23 (10.4%) were grade 1, 11 (5.0%) were grade 2, and 0 were grade 3. In group O, 351 (84.4%) of the 416 pedicle screw placements were categorized as grade 0, 52 (12.5%) were grade 1, 13 (3.1%) were grade 2, and 0 were grade 3. Statistical analysis showed no significant difference in the prevalence of grade 2.3 perforations between groups C and O. The time to position one screw, including registration, was 10.9±3.2 minutes in group C, but was significantly decreased to 5.4±1.1 minutes in group O. Conclusions O-arm-based navigation facilitates pedicle screw insertion as accurately as conventional CT-based navigation. The use of O-arm-based navigation successfully reduced the time, demonstrating advantages in the safety and accuracy of pedicle screw placement for scoliotic surgery.

Tumor Necrosis Factor-α-Immunoreactive Cells in Nucleus Pulposus in Adolescent Patients With Lumbar Disc Herniation

Study Design. Immunohistochemistry for tumor necrosis factor (TNF)–&agr; in nucleus pulposus of adolescent patients with lumbar disc herniation. Objective. To examine whether an inflammatory cytokine is expressed in the nucleus pulposus of adolescent patients with lumbar disc herniation. Summary of Background Data. TNF&agr; is thought to play a crucial role in the radicular pain caused by lumbar disc herniation in adult patients. However, the expression of TNF&agr; in the nucleus pulposus of adolescent patients with lumbar disc herniation has not been explored. Methods. Five samples of nucleus pulposus from adolescent patients with lumbar disc herniation (age, 12–16 yr; n = 5) or controls requiring surgery for other back problems (age, 12–16 yr; n = 4; nonpainful scoliosis) were harvested during surgery. Nucleus pulposus specimens were immunostained using TNF&agr; antibodies and immunostained cells in the nucleus pulposus were counted. We compared the expression of TNF&agr; between the 2 groups. Results. In patients with lumbar disc herniation, more TNF&agr;-immunoreactive cells were seen in the nucleus pulposus in comparison with patients with nonpainful scoliosis (P < 0.01). Conclusion. The results suggest that TNF&agr; may play a role in adolescent patients with lumbar disc herniation. The TNF&agr; expression may be related with disc degeneration and pain in adolescent patients with lumbar disc herniation.