Saihu Mao

Genome-wide association study identifies new susceptibility loci for adolescent idiopathic scoliosis in Chinese girls

Adolescent idiopathic scoliosis (AIS) is a structural deformity of the spine affecting millions of children. As a complex disease, the genetic aetiology of AIS remains obscure. Here we report the results of a four-stage genome-wide association study (GWAS) conducted in a sample of 4,317 AIS patients and 6,016 controls. Overall, we identify three new susceptibility loci at 1p36.32 near AJAP1 (rs241215, Pcombined=2.95 × 10−9), 2q36.1 between PAX3 and EPHA4 (rs13398147, Pcombined=7.59 × 10−13) and 18q21.33 near BCL-2 (rs4940576, Pcombined=2.22 × 10−12). In addition, we refine a previously reported region associated with AIS at 10q24.32 (rs678741, Pcombined=9.68 × 10−37), which suggests LBX1AS1, encoding an antisense transcript of LBX1, might be a functional variant of AIS. This is the first GWAS investigating genetic variants associated with AIS in Chinese population, and the findings provide new insight into the multiple aetiological mechanisms of AIS.

A promoter polymorphism of neurotrophin 3 gene is associated with curve severity and bracing effectiveness in adolescent idiopathic scoliosis.

Study Design. A genetic association study to comprehensively investigate variations of neurotrophin 3 (NTF3) gene polymorphisms in a Chinese Han population. Objective. To explore whether the NTF3 gene polymorphisms are associated with the susceptibility, curve severity, or bracing effectiveness of adolescent idiopathic scoliosis (AIS). Summary of Background Data. Scoliosis has developed in mice with NTF3 deficiency in previous studies. Increased expression of NTF3 mRNA was detected in the paravertebral muscle in AIS. Moreover, linkage study has defined a novel AIS locus on chromosome 12p while NTF3 gene is located exactly in this interval. All evidence indicates a potential role of NTF3 in the pathogenesis of AIS. As for brace treatment of AIS, continuous sensory stimulation caused by an orthosis could help awareness of body misalignment and trigger curve correction through postural reflex. While NTF3 gene is tightly associated with proprioceptive feedback mechanism to adjust postural control, we hypothesized NTF3 as a potential candidate gene associated with the bracing effectiveness. Methods. A total of 362 AIS patients and 377 age-matched healthy controls were recruited. Two single-nucleotide polymorphisms (SNPs) were selected on the basis of the Chinese data from the HapMap project, and genotyping was carried out by polymerase chain reaction–restriction fragment length polymorphism for each SNP, respectively. Case-control study and case-only study were performed to define the contribution of NTF3 gene polymorphisms to predisposition and disease severity of AIS. Another subgroup of 120 skeletally immature AIS patients who received continuous brace treatment for minimal 2 years was genotyped, and bracing effectiveness was assessed to determine its association with NTF3 gene polymorphisms. Results. The genotype and allele frequency distribution were similar between AIS and normal control for these 2 SNPs (&khgr;2 test: P > 0.05). For SNP rs11063714 in the promoter region of NTF3 gene, AIS patients with AA genotype showed significantly lower mean maximum Cobb angle than the patients with AG or GG genotypes (analysis of variance: P = 0.008). In addition, skeletally immature bracing AIS patients with AA genotype possessed significantly higher successful ratio of brace treatment compared with GG genotype (&khgr;2 test: P = 0.043). For SNP rs1805149, no significant association with predisposition or curve severity was detected. Conclusion. The NTF3 gene polymorphisms are not associated with the occurrence of AIS, but the promoter polymorphism (rs11063714) is associated with the curve severity, implicating an alleviating role of NTF3 in the curve progression of AIS. In addition, the promoter polymorphism is also associated with brace responsiveness. These findings indicated that NTF3 gene might be a disease-modifying gene of AIS.

A promoter polymorphism of tissue inhibitor of metalloproteinase-2 gene is associated with severity of thoracic adolescent idiopathic scoliosis.

Study Design. A genetic association study of the tissue inhibitor of metalloproteinase-2 (TIMP-2) gene with adolescent idiopathic scoliosis (AIS) in a Chinese population. Objective. To determine whether a promoter polymorphism of the TIMP-2 gene correlates with the occurrence and curve severity of AIS patients. Summary of Background Data. Previous studies have suggested that genetic factors play an important role in the etiology of AIS. The relative anterior spinal column overgrowth due to abnormal endochondral ossification has been considered to be a significant factor in the etiopathogenesis of AIS. The specific role of matrix metalloproteases (MMPs) and their activity inhibitors, TIMPs, during endochondral ossification has been documented. The TIMP-2 is the major TIMP expressed during bone development and is located in one of the chromosomal regions linked to AIS. Therefore, the TIMP-2 gene is a potential candidate gene for AIS. Methods. This study included a total of 570 female AIS patients, who were divided into 2 groups according to curve patterns. Of them, 354 patients with right thoracic curve were in group A (326 cases with Lenke 1 type and 28 cases with Lenke 2 type), whereas 216 patients with a single lumbar curve were in group B (216 cases with Lenke 5 type). A total of 210 age-matched healthy girls were recruited as normal controls. One single-nucleotide polymorphism, −418G/C (rs8179090), in the promoter region was selected for the TIMP-2 gene. Genotyping was performed by polymerase chain reaction–restriction fragment length polymorphism in each group. Results. No significant differences of genotype and allele frequency distribution were found between AIS patients and normal controls either in group A or in group B. The frequency of C allele was significantly higher in patients with Cobb angle 40° or more than in those with Cobb angle less than 40° in group A (P < 0.05), while this difference was not noted in group B (P > 0.05). Among the patients who reached skeletal maturity without any interference of natural history, a significantly higher average maximum Cobb angle was found in patients with C allele than in those without C allele in group A (P < 0.05). However, in group B, the mean maximum Cobb angle was similar between patients with different genotypes in both cases with left-side curves and cases with right-side curves (P > 0.05). Furthermore, for the patients whose values of thoracic kyphosis were recorded, those with C allele had smaller average thoracic kyphosis than those without C allele in group A (P < 0.05). However, such significant difference was not observed in group B. Conclusion. The single-nucleotide polymorphism SNP-418G/C (rs8179090) in the promoter region of the TIMP-2 gene was not associated with the occurrence of AIS. However, it may predict curve severity of thoracic AIS. Hence, the TIMP-2 gene is a disease-modifier gene of thoracic AIS.

An updated analysis of pubertal linear growth characteristics and age at menarche in ethnic Chinese

Concerns regarding change in the onset and tempo of pubertal growth in ethnic Chinese have posed a need for current information on growth characteristics. This study is to update the normative data of pubertal linear growth characteristics and distribution of age at menarche in healthy Chinese adolescents.

Association between genetic determinants of peak height velocity during puberty and predisposition to adolescent idiopathic scoliosis.

Study Design. An association study to comprehensively clarify variations of genetic determinants of peak height velocity (PHV) during puberty in adolescent idiopathic scoliosis (AIS). Objective. To investigate whether the genetic determinants of timing and magnitude of PHV during puberty are associated with the susceptibility or curve progression of the female patients with AIS. Summary of Background Data. An involvement of abnormal pubertal growth pattern in the etiopathogenesis of AIS has been implicated in previous studies. However, there is no clear consensus on the anthropometric variations of stature or growth rate. The recent advance in the longitudinally identified genetic determinants of PHV offers new opportunities to facilitate analysis of the association of pubertal growth with the susceptibility or curve severity of AIS. Methods. A gene-based association study was conducted using 9 single nucleotide polymorphisms (SNPs) in or near SOCS2, SF3B4/SV2A, C17orf67, CABLES1, DOT1L, CDK6, C6orf106, and LIN28B with confirmed association with PHV, peak growth age, or adult height. A total of 500 patients with AIS and 494 age-matched healthy controls were genotyped using the PCR-based Invader assay. Case-control study and case-only study were performed to define the contribution of the 9 SNPs to predisposition and curve severity of AIS. Results. Strong associations between rs12459350 in DOT1L, rs4794665 in C17orf67, and susceptibility of AIS were found, with the PHV increasing allele G of rs12459350 and PHV/adult height increasing allele A of rs4794665 both being significant predisposition alleles of AIS (P = 0.001 for rs12459350, odds ratio = 1.16, 95% confidence interval = 1.06–1.27; P = 0.006 for rs4794665, odd ratio = 1.33, 95% confidence interval = 1.09–1.62). None of the genotyped SNPs was associated with curve severity in patients with AIS. Conclusion. Polymorphisms of the rs4794665 in C17orf67 and rs12459350 in DOT1L were associated with combined predisposition to AIS susceptibility and higher pubertal PHV, which strongly mirrored the anthropometric findings of taller pubertal stature and accelerated growth rate described in AIS.

Different proximal thoracic curve patterns have different relative positions of esophagus to spine in adolescent idiopathic scoliosis: a computed tomography study.

Study Design. A computed tomography (CT) study. Objective. To evaluate the changed relative positions of esophagus in proximal thoracic (PT) curves of adolescent idiopathic scoliosis (AIS) patients and analyze the potential risks of esophageal injuries from thoracic pedicle screw (TPS) insertion. Summary of Background Data. Translation and rotation of the vertebrae could lead to altered relative positions of surrounding vital structures in AIS patients. The changed positions of aorta and spinal cord in main thoracic (MT) curve have been comprehensively investigated; however, no studies have analyzed the relative position of esophagus in PT curve. Methods. Twenty patients with complete proximal thoracic (CPT group) curve, 22 patients with fractional proximal thoracic (FPT group) curve, and 14 normal patients with a straight spine (normal group) were included. Axial CT images from T2 to T5 at the midvertebral body level were obtained to evaluate esophagus-vertebral angle (EVA, defined as 0° when the esophagus was located directly lateral to the left, 90° when strictly anterior, and 180° when directly lateral to the right). The percentages of esophagus in the direction of screw passage were calculated to analyze potential risks of esophageal injuries during TPS insertion. Results. EVA in the FPT group was significantly smaller than that in the normal group (P < 0.05), whereas EVA in the CPT group was significantly greater than that in the normal group (P < 0.05) at each level. The esophagus was located approximately anterior to the vertebral body in the normal group but shifted anterolaterally to the right in the CPT group and anterolaterally to the left in the FPT group. The esophagus was at a high risk of injury with right anterior penetrated TPS in the CPT group and was at a high risk of injury with left anterior penetrated TPS in the FPT group. Conclusion. Different anatomic patterns of PT curves could cause different altered positions of esophagus relative to spine and result in different potential risks of esophageal injuries during TPS insertion. Spine surgeons should choose appropriate pedicle screw length to avoid anterior cortical perforation in the PT region of AIS patients.

Outcomes and predictors of brace treatment for girls with adolescent idiopathic scoliosis.

Objective:  To evaluate the effectiveness and to identify the predictive factors of standardized brace treatment for girls with adolescent idiopathic scoliosis (AIS).

Mechanisms, Predisposing Factors, and Prognosis of Intraoperative Vertebral Subluxation During Pedicle Subtraction Osteotomy in Surgical Correction of Thoracolumbar Kyphosis Secondary to Ankylosing Spondylitis

Study Design. A retrospective study. Objective. To analyze the mechanisms, predisposing factors, and prognosis of the intraoperative vertebral subluxation (VS) during pedicle subtraction osteotomy (PSO) for thoracolumbar kyphosis secondary to ankylosing spondylitis (AS). Summary of Background Data. VS is one of the most daunting challenges that surgeons encounter during PSO closure, especially in patients with AS with ankylosed and mostly osteoporotic spine. Unfortunately, there is a paucity of research designed to conceptualize the mechanisms, predisposing factors, and discuss the complication-avoidance strategies and prognosis. Methods. A retrospective single-center review was performed for a consecutive series of 153 patients with AS with rigid thoracolumbar kyphosis who underwent one-level PSO from April 2000 to December 2013. The incidence of the VS at the level of PSO during correction was analyzed and the potential causative factors were investigated. Results. VS occurred in six patients with the incidence being 3.9% in this patient cohort. The predisposing factors were (1) early fracture of the anterior cortex of the osteotomized vertebra (OV); (2) excessive decancellation from vertebral body causing parallel collapse of the vertebral column with significant loss of the ability to create local lordosis; (3) improper manual osteoclasis due to insufficient decancellation of the OV; and (4) inappropriate application of cantilever technique and concomitant long instrumentation. The early surgical complication involved one patient with cerebrospinal fluid leakage at the osteotomized site, but no devastating neurological deficits. During follow-up, bone healing and adaptive vertebral remodeling with no rod breakage were observed for all these six patients. Conclusion. Intraoperative VS was a rare occurrence associated with inappropriate manual manipulation of osteotomy, gap closure, and rod insertion. Neurological complication was a potential risk, but could be well prevented with extensive laminectomy and emergency actions favoring partial subluxation reduction. Moreover, adaptive bone remodeling and fusion at the level of VS ensured the maintenance of kyphosis correction and avoidance of instrumentation failure. Level of Evidence: 4

Investigation of the 53 Markers in a DNA-Based Prognostic Test Revealing New Predisposition Genes for Adolescent Idiopathic Scoliosis.

Study Design. A genetic association study of 53 single nucleotide polymorphisms (SNPs) with adolescent idiopathic scoliosis (AIS). Objective. To explore new predisposition genes of AIS in Chinese Han population Summary of Background Data. A panel of 53 SNPs were reported to be associated with curve severity of AIS. However, there is still a lack of knowledge concerning the association of these SNPs with the susceptibility of AIS in the Chinese Han population. Methods. A gene-based association study was conducted by genotyping the 53 SNPs of a prognostic test. DNA samples of 990 female patients with AIS and 1188 age-matched healthy controls were analyzed using the polymerase chain reaction-based Invader assay. The &khgr;2 test was carried out to compare the differences of genotype and allele distributions between patients with AIS and healthy controls. Results. A total of 4 SNPs were found to present significant differences in allele or genotype frequencies between the 2 groups. Compared with normal controls, patients were found to have significantly higher allele G of rs12618119 and allele A of rs9945359. Besides, patients were found to have significantly lower allele T of rs4661748 and allele C of rs4782809 than the normal controls. BIN1, CDH13, SETBP1, and SPATA21 genes could be associated with the susceptibility of AIS. Conclusion. Four new predisposition genes of AIS were identified on the basis of a large-scale case-control study. Putting all these findings together, it suggests that AIS is a multifactorial disease possibly involving different pathways such as development of central neural system and bone formation. Further studies exploring more predisposition gene are essential to illustrate the etiology of AIS and to guide the prevention or prognosis of the disease. Level of Evidence: 3

Change in Abdominal Morphology After Surgical Correction of Thoracolumbar Kyphosis Secondary to Ankylosing Spondylitis: A Computed Tomographic Study.

Study Design. A computed tomographic study. Objective. To investigate the change in abdominal morphology in surgically treated patients with ankylosing spondylitis (AS) and thoracolumbar kyphosis. Summary of Background Data. Severe thoracolumbar kyphosis in patients with AS exerts pressure on the abdominal cavity and subsequently causes intra-abdominal complications. Several spinal osteotomy techniques have been widely used to correct AS-related thoracolumbar kyphosis. To date, the changed abdominal morphology in patients with AS undergoing surgical correction of thoracolumbar kyphosis has not been addressed. Methods. A total of 29 patients with AS undergoing lumbar pedicle subtraction osteotomy for correction of thoracolumbar kyphosis were retrospectively reviewed. Computed tomographic scans of the spine were used to measure the longitudinal, transverse, and anterior–posterior diameters of the abdominal cavity. Furthermore, the abdominal cavity was considered as an ellipsoid structure, thereby allowing calculation of its volume. Radiographical evaluations included global kyphosis (GK), thoracic kyphosis, lumbar lordosis (LL), and angle of fusion levels (AFL). Results. The longitudinal diameter of abdominal cavity significantly increased (P < 0.01), whereas the transverse and anterior–posterior diameters of the abdominal cavity did not change, postoperatively (P > 0.05). Significant changes in GK, LL, and AFL were observed (P < 0.01). The abdominal cavity volume (ACV) increased by an average of 652 mL. The change in ACV was significantly correlated with the changes in GK (r = 0.453, P = 0.014), LL (r = 0.42, P = 0.023), and AFL (r = 0.388, P = 0.037). Conclusion. The increased ACV after correction of thoracolumbar kyphosis was quantitatively confirmed by this study. Thus, the improvement in digestive function after correction of thoracolumbar kyphosis secondary to AS, which has been previously documented, may be because of an increase in ACV. Moreover, spine surgeons should be aware of the potential risk for the development of abdominal complications caused by the lengthening of longitudinal diameter of the abdominal cavity. Level of Evidence: 3