Noncanonical structural requirements of neurofibromin SUMOylation reveal a folding-deficiency of several pathogenic mutants

Neurofibromin (Nf1) is a large multidomain protein encoded by the tumour-suppressor gene NF1. NF1 is mutated in a frequently occurring genetic disease, neurofibromatosis type I, and in various cancers. The best described function of Nf1 is its Ras-GTPase activity, carried out by its GAP-related domain (GRD). SecPH, another structurally well-characterized domain of Nf1, is immediately adjacent to the GRD and interacts with lipids and proteins, thus connecting Nf1 to diverse signalling pathways. Here, we demonstrate, for the first time, that Nf1 and SecPH are substrates of the SUMO pathway. We identified a well-defined SUMOylation profile of SecPH and a main SUMOylation event on Lys1731 that appears to play a role in Ras-GAP activity. Our data allowed us to characterize a new set of pathogenic Nf1 missense mutants that exhibits a disrupted SUMOylation profile that may correlate with their unfolding. Accordingly, Lys1731 SUMOylation is mediated by a noncanonical structural motif, therefore allowing a read-out of SecPH conformation and folding status.

Efficacy and safety for combination of t-EMG with O-arm assisted pedicle screw placement in neurofibromatosis type I scoliosis surgery

Background Due to the characteristics of neurofibromatosis type I (NF-1) scoliosis, the precise placement of pedicle screws still remains to be a challenge. Triggered screw electromyography (t-EMG) has been proved to exhibit high sensitivity to identify mal-positioned pedicle screws, but no previous study assessed the combination of t-EMG with O-arm-assisted pedicle screw placement in NF-1 scoliosis surgery. Objective To evaluate efficacy and safety for combination of t-EMG with O-arm-assisted pedicle screw placement in NF-1 scoliosis surgery. Materials and methods From March 2018 to April 2020, sixty-five NF-1 scoliosis patients underwent t-EMG and O-arm-assisted pedicle screw fixation were retrospectively reviewed. The channel classification system was applied to classify the pedicle morphology based on pedicle width measurement by preoperative computed tomography scans. The minimal t-EMG threshold for screw path inspection was used as 8 mA, and operative screw redirection was also recorded. All pedicle screws were verified using a second intraoperative O-arm scan. The correlation between demographic and clinical data with amplitude of t-EMG were also analyzed. Results A total of 652 pedicle screws (T10-S1) in 65 patients were analyzed. The incidence of an absent pedicle (channel classification type C or D morphology) was 150 (23%). Overall, abnormal t-EMG threshold was identified in 26 patients with 48 screws (7.4%), while 16 out of the 48 screws were classified as G0, 14 out of the 48 screws were classified as G1, and 18 out of the 48 screws were classified as G2. The screw redirection rate was 2.8% (18/652). It showed that t-EMG stimulation detected 3 unacceptable mal-positioned screws in 2 patients (G2) which were missed by O-arm scan. No screw-related neurological or vascular complications were observed. Conclusions Combination of t-EMG with O-arm-assisted pedicle screw placement was demonstrated to be a safe and effective method in NF-1 scoliosis surgery. The t-EMG could contribute to detecting the rupture of the medial wall which might be missed by O-arm scan. Combination of t-EMG with O-arm could be recommended for routine use of screw insertion in NF-1 scoliosis surgery.

Case Report: A Synonymous Mutation in NF1 Located at the Non-canonical Splicing Site Leading to Exon 45 Skipping

Synonymous mutations are generally considered non-pathogenic because it did not alter the amino acids of the encoded protein. Publications of the associations between synonymous mutations and abnormal splicing have increased recently, however, not much observations available described the synonymous mutations at the non-canonical splicing sites leading to abnormal splicing. In this pedigree, the proband was diagnosed Neurofibromatosis type I due to the presence of typical cafe’ au lait macules and pectus carinatum. Whole-exome sequencing identified a synonymous mutation c.6795C > T (p.N2265N) of the NF1 gene which was located at the non-canonical splicing sites. Reverse transcription polymerase chain reaction followed by Sanger sequencing was carried out, and the skipping of exon 45 was observed. Therefore, the pathogenicity of the synonymous mutation c.6795C > T was confirmed. Our finding expanded the spectrum of pathogenic mutations in Neurofibromatosis type I and provided information for genetic counseling.

Neurofibromatosis Type I and Stromal Tumor with a Multiple Digestive Localization

Neurofibromatosis type I (NF1) is also known as von Recklinghausen disease. It is a genetic disorder that affects the growth and development of nerve cell tissue, which is characterized by a multisystem disorder and an increased risk for cancer. The incidence of gastroduodenal stromal tumor during Recklinghausen disease can reach 35% in autopsies and 5% in clinical cases. In our case, the diagnosis of neurofibromatosis type I was made in a middle-aged women initially diagnosed with a pancreaticoduodenal tumor.