Genetic insights into the 'sandwich fusion' subtype of Klippel-Feil syndrome: novel FGFR2 mutations identified by 21 cases of whole-exome sequencing.

BACKGROUND: Klippel-Feil syndrome (KFS) is a rare congenital disorder characterized by the fusion of two or more cervical vertebrae during early prenatal development. This fusion results from a failure of segmentation during the first trimester. Although six genes have previously been associated with KFS, they account for only a small proportion of cases. Among the distinct subtypes of KFS, "sandwich fusion" involving concurrent fusion of C0-1 and C2-3 vertebrae is particularly noteworthy due to its heightened risk for atlantoaxial dislocation. In this study, we aimed to investigate novel candidate mutations in patients with "sandwich fusion." METHODS: We collected and analyzed clinical data from 21 patients diagnosed with "sandwich fusion." Whole-exome sequencing (WES) was performed, followed by rigorous bioinformatics analyses. Our focus was on the six known KFS-related genes (GDF3, GDF6, MEOX1, PAX1, RIPPLY2, and MYO18). Suspicious mutations were subsequently validated through in vitro experiments. RESULTS: Our investigation revealed two novel exonic mutations in the FGFR2 gene, which had not previously been associated with KFS. Notably, the c.1750A > G variant in Exon 13 of FGFR2 was situated within the tyrosine kinase domain of the protein, in close proximity to several established post-translational modification sites. In vitro experiments demonstrated that this certain mutation significantly impacted the function of FGFR2. Furthermore, we identified four heterozygous candidate variants in two genes (PAX1 and MYO18B) in two patients, with three of these variants predicted to have potential clinical significance directly linked to KFS. CONCLUSIONS: This study encompassed the largest cohort of patients with the unique "sandwich fusion" subtype of KFS and employed WES to explore candidate mutations associated with this condition. Our findings unveiled novel variants in PAX1, MYO18B, and FGFR2 as potential risk mutations specific to this subtype of KFS.

Preoperative radiological indicators for prediction of difficult laryngoscopy in patients with atlantoaxial dislocation.

Background: Difficult airway remains a great challenge in patients with atlantoaxial dislocation (AAD). Preoperative evaluation and reliable prediction are required to facilitate the airway management. We aimed to screen out reliable radiological indicators for prediction of difficult laryngoscopy in patients with AAD. Methods: A retrospective nested case-control study within a single center longitudinal AAD cohort was conducted to investigate the radiological indicators. All the patients with difficult laryngoscopy from 2010 to 2021 were enrolled as the difficult laryngoscopy group. Others in the cohort without difficult laryngoscopy were randomly selected as the non-difficult laryngoscopy group by individually matching with the same gender, same surgery year, and similar age (±5 years) at a ratio of 6:1. Radiological data on preoperative lateral X-ray images between the two groups were compared. Bivariate logistic regression model was applied to screen out the independent predictive indicators and calculate the odds ratios of indicators associated with difficult laryngoscopy. Receiver operating characteristic curve and area under the curve (AUC) were used to describe the discrimination ability of indicators. Results: A total of 154 patients were finally analyzed in this study. Twenty-two patients with difficult laryngoscopy and matched with 132 controls. Four radiological parameters showed significant difference between the two groups. Among which, ΔC1C2D (the difference of the distance between atlas and axis in the neutral and extension position), owned the largest AUC. Conclusions: ΔC1C2D could be a valuable radiologic predictor for difficult laryngoscopy in patients with AAD.

Overstrain on the longitudinal band of the cruciform ligament during flexion in the setting of sandwich deformity at the craniovertebral junction: a finite element analysis.

BACKGROUND CONTEXT: In the setting of "sandwich deformity" (concomitant C1 occipitalization and C2-3 nonsegmentation), the C1-2 joint becomes the only mobile joint in the craniovertebral junction. Atlantoaxial dislocation develops earlier with severer symptoms in sandwich deformity, which has been hypothesized to be due to the repetitive excessive tension in the ligaments between C1 and C2. PURPOSE: To elucidate whether and how the major ligaments of the C1-2 joint are affected in sandwich deformity, and to find out the ligament most responsible for the earlier development and severer symptoms of atlantoaxial dislocation in sandwich deformity. STUDY DESIGN: A finite element (FE) analysis study. METHODS: A three-dimensional FE model from occiput to C5 was established using anatomical data from a thin-slice CT scan of a healthy volunteer. Sandwich deformity was simulated by eliminating any C0-1 and C2-3 segmental motion respectively. Flexion torque was applied, and the range of motion of each segment and the tension sustained by the major ligaments of C1-2 (including the transverse and longitudinal bands of the cruciform ligament, the alar ligaments, and the apical ligament) were analyzed. RESULTS: Tension sustained by the longitudinal band of the cruciform ligament and the apical ligament during flexion is significantly larger in the FE model of sandwich deformity. In contrast, tension in the other ligaments is not significantly changed in the sandwich deformity model compared with the normal model. CONCLUSIONS: Considering the importance of the longitudinal band of the cruciform ligament to the stability of the C1-2 joint, our findings implicate that the early onset, severe dislocation, and unique clinical manifestations of atlantoaxial dislocation in patients with sandwich deformity are mainly due to the enlarged force loaded on the longitudinal band of the cruciform ligament. CLINICAL SIGNIFICANCE: The enlarged force loaded on the longitudinal band of the cruciform ligament can add to its laxity and thus reducing its ability to restrict the cranial migration of the odontoid process. This is in accordance with our clinical experience that dislocation of the atlantoaxial joint in patients with sandwich deformity is mainly craniocaudal, which means severer cranial neuropathy, Chiari deformity, and syringomyelia, and more difficult surgical treatment.

Clinical and Surgical Characteristics of Patients with Atlantoaxial Dislocation in the Setting of Sandwich Fusion: A Case-Control Analysis of Over 500 Patients with Mid-Term to Long-Term Follow-up.

BACKGROUND: Patients with "sandwich" fusion (concomitant C1 occipitalization and C2-C3 nonsegmentation), a subtype of Klippel-Feil syndrome, are at particular risk for developing atlantoaxial dislocation (AAD). However, the clinical and surgical characteristics of AAD in patients with sandwich fusion have not been clearly defined. METHODS: A retrospective case-control study with a large sample size and a minimum 2-year follow-up was performed. From 2000 to 2018, 253 patients with sandwich AAD underwent a surgical procedure; these patients constituted the case group, and a matching number of patients with non-sandwich AAD were randomly selected to form the control group. Clinical data from electronic medical records and various imaging studies were analyzed and compared. The Japanese Orthopaedic Association (JOA) scale was used to evaluate neurological function. RESULTS: Patients with sandwich AAD, compared with patients with non-sandwich AAD, had symptom onset at a younger age (34.8 compared with 42.8 years; p < 0.001) and had a higher likelihood for myelopathy (87.4% compared with 74.7%; p < 0.001). Patients with sandwich AAD had a higher incidence of lower cranial nerve palsy (7.9% compared with 0.0%; p < 0.001), a lower preoperative JOA score (13.4 compared with 14.2; p < 0.001), and higher incidences of accompanying Type-I Chiari malformation (20.9% compared with 1.2%; p < 0.001) and syringomyelia (21.3% compared with 1.6%; p < 0.001). Finally, patients with sandwich AAD had higher likelihoods of undergoing transoral release (28.5% compared with 5.1%; p < 0.001) and use of salvage fixation techniques (34.4% compared with 6.3%; p < 0.001), and had lower postoperative results for the JOA score (14.9 compared with 15.9; p < 0.001) and improvement rate (43.8% compared with 58.2%; p < 0.001). CONCLUSIONS: Patients with sandwich AAD demonstrated distinct clinical manifestations. Versatility involving the use of various internal fixation techniques and transoral release procedures was frequently required in the surgical management of these patients, and meticulous and personalized preoperative planning would be of paramount importance. LEVEL OF EVIDENCE: Prognostic Level III . See Instructions for Authors for a complete description of levels of evidence.

Strategies to avoid internal carotid artery injury in "sandwich" atlantoaxial dislocation patients during surgery.

PURPOSE: To elucidate the anatomic relationship between the internal carotid artery (ICA) and the bony structures of the craniovertebral junction among "sandwich" atlantoaxial dislocation (AAD) patients, and to analyze the risks of injury during surgical procedures. METHODS: The distance from the medial wall of ICA to the midsagittal plane (D1), the shortest distance between the ICA wall and the anterior cortex of the lateral mass of atlas (LMA) (D2) on the most caudal and cranial levels of LMA and the angle (A) between the sagittal plane passing through the screw entry point of C1 lateral mass(C1LM) screw and the medial tangent line of the vessel passing through the entry point were measured. Besides, the location of ICA in front of the atlantoaxial vertebra was divided into 4 categories (Z1-Z4). RESULTS: There was a statistically difference between the male and female patients regarding D1, and the difference between D2 at level a and level b as well as angle A between the left and right sides were statistically different (p < 0.05). Ninety-two ICAs (57.5%) were anteriorly located in Z3, 50 (31.3%) were located in Z4, 17 were located in Z2, and only one ICA was located in Z1 in all 80 patients. CONCLUSIONS: In "sandwich" AAD patients, particular attention should be paid to excessively medialized ICA to avoid ICA injury during trans-oral procedures, and the risk of injuring the ICA with more cranially and medially angulated C1LM screw placement was relatively less during posterior fixation procedures. A novel classification of ICA location was used to describe the relationship between ICA and LMA.

Spinous Process Screw Fixation: A Salvage Technique in Subaxial Cervical Spinal Instrumentation.

OBJECTIVE: We sought to evaluate the feasibility and efficacy of spinous process screws in subaxial cervical fixation. METHODS: A retrospective study was conducted on 7 patients receiving posterior cervical instrumentation for subaxial fracture dislocation or atlantoaxial dislocation from 2014 to 2015. In addition, a biomechanical test was performed on 7 fresh-frozen cadaveric spine samples (from the occiput to C7) from healthy human subjects with no history of spinal trauma. And lastly, the potential trajectories of cervical spinous process screws of 100 adults were measured through axial computed tomography in order to establish the general rule for applicability of this technique. RESULTS: Rigid fixation and solid fusion were achieved in all 7 patients included in the study. Biomechanical test results revealed no significant difference between bilateral lateral mass screw fixation and the hybrid lateral mass and spinous process screw fixation constructs. Measurement of the potential trajectory suggested that spinous process screw fixation was a viable technique in the subaxial cervical spine in the general population. CONCLUSIONS: The spinous process screw fixation technique in the subaxial cervical spine is a viable and effective salvage option for patients in whom conventional posterior fixations were not sufficient and salvage, supplementary techniques were required.

A novel 3D-printed locking cage for anterior atlantoaxial fixation and fusion: case report and in vitro biomechanical evaluation.

BACKGROUND: Treatment of atlantoaxial dislocation is aimed at reduction and stabilization of the atlantoaxial joint. 3D printing refers to a process where additive manufacturing is achieved under precise computer control. Literature on its utilization in anterior atlantoaxial fixation and fusion is rare. This study is the first report on a 3D-printed locking cage used in the anterior procedure for atlantoaxial dislocation. METHODS: A middle-aged male in his 40s presented with weakness and numbness of his extremities for 3 years and could only walk slowly with assistance. Imaging studies revealed severe anterior migration of C1, irreducible atlantoaxial dislocation, and severe cervical-medullary compression. A preoperative plan consisting of trans-oral soft tissue release and fixation using tailor-designed 3D-printed cages was devised. Following fluoroscopic confirmation of reduction of the atlantoaxial joints, two customized 3D-printed cages made of titanium alloy were inserted into the bilateral facet joints, which were then locked by six screws into the lateral masses of C1 and C2. The microstructure of the inserted cages was optimized for improved biomechanical stability and enhanced osseo-integration, without the need for bone grafting. In addition, a biomechanical test was performed on seven human cadaveric specimens comparing the novel implant with the conventional C1 lateral mass-C2 pedicle screw construct in three modes of motion (flexion-extension, lateral bending, axial rotation). RESULTS: Improvement of neurologic function in the patient was evident immediately after surgery. He was able to walk independently 1 month post-operatively. At the 12-month follow-up, coronal reconstruction of CT demonstrated properly-positioned 3D-printed cages, evidence of osseo-integration at the bone-implant interface, and no subsidence or displacement of the implant. Eighteen months out of surgery, the mJOA score improved to 15, and lateral X-ray confirmed reduction of atlanto-axial dislocation. Additionally, the new construct provided strong fixation comparable to that conferred by conventional constructs as there was no significant difference observed between the two groups in all three directions of motion. CONCLUSIONS: The novel implant represents a new option in the treatment of irreducible atlantoaxial dislocation. It can provide strong anterior support for solid fixation and fusion with a low profile and a microstructure that obviates the need for bone grafting.

Vertebral Artery Variations at the Craniovertebral Junction in "Sandwich" Atlantoaxial Dislocation Patients.

OBJECTIVE: To summarize the vertebral artery (VA) pattern of 96 "sandwich" atlantoaxial dislocation (AAD) patients and to describe the strategies of reducing the injury of VA during surgery. METHODS: From 2009 to 2020, we retrospectively reviewed the 3-dimensional computed tomography angiography data of 96 AAD patients combined with atlas occipitalization and C2-3 fusion, which were diagnosed as "sandwich" AAD and 96 patients as control group patients who were without atlas occipitalization, C2-3 fusion and any other cervical bone deformity at our institution. The variations of each side of VA were described in 3 different parts (C0-1, C1-2, and C2-3) according to the characteristics of the 3-part pathological structures in "sandwich" subgroup. RESULTS: One hundred ninety-two sides of VAs in every group of patients were analyzed and every VA was described separately at 3 different level regions. There were different variations in these 3 different regions: 4 variations in the upper fusion region, 5 variations in the sandwiched region, and 6 variations in the lower fusion region in sandwich AAD patients. And the rate of VA deformity in sandwich AAD patients was much higher and more types of VA variations existed. CONCLUSION: In "sandwich" AAD patients, deformities of vertebral arteries in craniovertebral junction are more common, and the same VA may have deformities at different levels that severely affect surgical procedures. Therefore, preoperative imaging examination of VA for "sandwich" AAD patients is vital of guiding surgeons to avoid injury of VA during surgery.

Atlantoaxial dislocation with congenital "sandwich fusion" in the craniovertebral junction: a retrospective case series of 70 patients.

BACKGROUND: In the setting of congenital C1 occipitalization and C2-3 fusion, significant strain is placed on the atlantoaxial joint. Vertebral fusion both above and below the atlantoaxial joint (i.e., a "sandwich") creates substantial instability. We retrospectively report on a case series of "sandwich fusion" atlantoaxial dislocation (AAD), describing the associated clinical characteristics and detailing surgical treatment. To the best of our knowledge, the present study is the largest investigation to date of this congenital subgroup of AAD. METHODS: Seventy consecutive patients with sandwich fusion AAD, from one senior surgeon, were retrospectively reviewed. The clinical features and the surgical treatment results were assessed using descriptive statistics. No funding sources or potential conflict of interest-associated biases exist. RESULTS: The mean patient age was 42.2 years (range: 5-77 years); 36 patients were male, and 34 were female. Fifty-eight patients (82.9%) had myelopathy, with Japanese Orthopaedic Association (JOA) scores ranging 4-16 (mean: 12.9). Cranial neuropathy was involved in 10 cases (14.3%). The most common presentation age group was 31 to 40 years (24 cases, 34.3%). Radiological findings revealed brainstem and/or cervical-medullar compression (58 cases, 82.9%), syringomyelia (16 cases, 22.9%), Chiari malformation (12 cases, 17.1%), cervical spinal stenosis (10 cases, 14.3%), high scapula deformity (1 case, 1.4%), os odontoideum (1 case, 1.4%), and dysplasia of the atlas (1 case, 1.4%). Computed tomography angiography was performed in 27 cases, and vertebral artery (VA) anomalies were identified in 14 cases (51.9%). All 70 patients underwent surgical treatment, without spinal cord or VA injury. Four patients (5.7%) suffered complications, including 1 wound infection, 1 screw loosening, and 2 cases of bulbar paralysis. In the 58 patients with myelopathy, the mean JOA score increased from 12.9 to 14.5. The average follow-up time was 50.5 months (range: 24-120 months). All 70 cases achieved solid atlantoaxial fusion at the final follow-up. CONCLUSIONS: Sandwich fusion AAD, a unique subgroup of AAD, has distinctive clinical features and associated malformations such as cervical-medullar compression, syringomyelia, and VA anomalies. Surgical treatment of AAD was associated with myelopathy improvement and minimal complication occurrence.

"Sandwich Deformity" in Klippel-Feil syndrome: A "Full-Spectrum" presentation of associated craniovertebral junction abnormalities.

Klippel-Feil syndrome (KFS) is defined as congenital fusion of two or more cervical vertebrae resulting from a segmentation failure in the developing spine. According to Samartzis et al., the most commonly fused segments are found at C2/3 (74.1%) and C6/7 (70.4%). In patients with C2/3 fusion, especially when there is additional C1 occipitalization, several secondary anomalies including atlantoaxial dislocation (AAD), basilar invagination (BI), Chiari malformation, and syringomyelia can be identified. In this report, we present a case of a 12-year-old patient with C2/3 and occipitalization and a "Full-Spectrum" presentation of associated CVJ abnormalities including C0/1 fusion, AAD, BI, Chiari malformation, syringomyelia, myelopathy and cranial neuropathy received neurological decompression of the cervico-medullary junction by posterior reduction of the AAD and reconstruction of her CVJ using an unconventional hybrid construct due to a high-riding right vertebral artery in C2. To our knowledge, her "Full-Spectrum" presentation may include the most categories of concomitant abnormalities in the literature. In addition, She received neurological decompression of the cervico-medullary junction using an unconventional hybrid construct due to a high-riding vertebral artery in C2. Three months after the surgery, all of her symptoms recovered significantly. Neither Chiari malformation nor syringomyelia could be identified by MRI two years after the surgery. At the last follow-up (4 years), the patient became completely asymptomatic.

Treatment of atlantoaxial dislocations among patients with cervical osseous or vascular abnormalities utilizing hybrid techniques.

OBJECTIVE Most cervical fixations for atlantoaxial dislocation (AAD) are bilateral and symmetric; however, in the setting of osseous and vascular deformity at the craniovertebral junction, asymmetrical and hybrid fixations are used as "salvage" techniques. Because of the rarity of these cases, hybrid cervical fixations for AAD have not been fully explored. The aim of this study was to evaluate the clinical feasibility and outcomes of posterior hybrid cervical fixations for AAD. METHODS Twenty-one AAD cases were retrospectively studied; 18 had cervical myelopathy with Japanese Orthopaedic Association (JOA) scores ranging from 9 to 16 (mean 13.5). Hybrid fixation techniques included unilateral pedicle screws, transarticular screws, C-2 laminar screws, cervical lateral mass screws, and spinous process screws. During the same period, 82 AAD cases, treated using symmetric traditional fixations, were analyzed as controls. RESULTS Atlantoaxial fixation was performed in 11 cases, while occiput-cervical fixation was used in 10 cases. All cases achieved solid osseous fusion. Anatomical reduction was achieved in 20 cases (95.2%). All 18 cases with myelopathy showed postoperative improvement, with JOA scores ranging from 13 to 17 (mean 15.5). Three cases (14.2%) experienced complications, including delayed wound healing, CSF leakage, and fixation loosening. Hybrid fixation techniques showed significantly greater estimated blood loss when compared with controls (208.1 ± 19.30 ml vs 139.63 ± 8.75 ml, p = 0.001). Operative duration (125.38 ± 6.29 min vs 119.41 ± 3.77 min, p = 0.464), complication rates (14.3% vs 4.9%, p = 0.148), and JOA improvement rates (61% ± 7% vs 49% ± 4%, p = 0.161) showed no significant differences. CONCLUSIONS For ADD with osseous or vascular deformity, posterior cervical reduction and stabilization can be achieved using hybrid techniques, resulting in comparable clinical results to symmetric traditional fixation.