Moyamoya disease patient mutations in the RING domain of RNF213 reduce its ubiquitin ligase activity and enhance NFκB activation and apoptosis in an AAA+ domain-dependent manner

Moyamoya disease (MMD) is a cerebrovascular disease characterized by progressive occlusion of the internal carotid arteries. Genetic studies originally identified RNF213 as an MMD susceptibility gene that encodes a large 591 kDa protein with a functional RING domain and dual AAA+ ATPase domains. As the functions of RNF213 and its relationship to MMD onset are unknown, we set out to characterize the ubiquitin ligase activity of RNF213, and the effects of MMD patient mutations on these activities and on other cellular processes. In vitro ubiquitination assays, using the RNF213 RING domain, identified Ubc13/Uev1A as a key ubiquitin conjugating enzyme that together generate K63-linked polyubiquitin chains. However, nearly all MMD patient mutations in the RING domain greatly reduced this activity. When full-length proteins were overexpressed in HEK293T cells, patient mutations that abolished the ubiquitin ligase activities conversely enhanced nuclear factor κB (NFκB) activation and induced apoptosis accompanied with Caspase-3 activation. These induced activities were dependent on the RNF213 AAA+ domain. Our results suggest that the NFκB- and apoptosis-inducing functions of RNF213 may be negatively regulated by its ubiquitin ligase activity and that disruption of this regulation could contribute towards MMD onset

Increased abundance of Ruminococcus gnavus in gut microbiota is associated with moyamoya disease and non-moyamoya intracranial large artery disease

Moyamoya disease (MMD) is a rare cerebrovascular disease endemic in East Asia. The p.R4810K mutation in RNF213 gene confers a risk of MMD, but other factors remain largely unknown. We tested the association of gut microbiota with MMD. Fecal samples were collected from 27 patients with MMD, 7 patients with non-moyamoya intracranial large artery disease (ICAD) and 15 control individuals with other disorders, and 16S rRNA were sequenced. Although there was no difference in alpha diversity or beta diversity between patients with MMD and controls, the cladogram showed Streptococcaceae was enriched in patient samples. The relative abundance analysis demonstrated that 23 species were differentially abundant between patients with MMD and controls. Among them, increased abundance of Ruminococcus gnavus > 0.003 and decreased abundance of Roseburia inulinivorans < 0.002 were associated with higher risks of MMD (odds ratio 9.6, P = 0.0024; odds ratio 11.1, P = 0.0051). Also, Ruminococcus gnavus was more abundant and Roseburia inulinivorans was less abundant in patients with ICAD than controls (P = 0.046, P = 0.012). The relative abundance of Ruminococcus gnavus or Roseburia inulinivorans was not different between the p.R4810K mutant and wildtype. Our data demonstrated that gut microbiota was associated with both MMD and ICAD

頚動脈管の経時的狭小化と内頚動脈陰性リモデリング評価への応用

京都大学新制・課程博士博士(医学)甲第23807号医博第4853号新制||医||1058(附属図書館)京都大学大学院医学研究科医学専攻(主査)教授 湊谷 謙司, 教授 YOUSSEFIAN Shohab, 教授 石見 拓学位規則第4条第1項該当Doctor of Medical ScienceKyoto UniversityDFA

Morphological Patterns of the Anterior Median Fissure in the Cervical Spinal Cord Evaluated by Computed Tomography After Myelography

Objective Computed tomography following myelography (CTM) revealed an unusual flow of contrast dye into the anterior median fissure (AMF) in a patient with cervical spondylotic myelopathy. Since then, several AMF configurations have been observed on CTM. Therefore, we evaluated morphological patterns of the AMF on CTM and investigated the significance and mechanisms of contrast dye flow into the AMF. Methods Morphological patterns of the AMF on CTM were examined in 79 patients. Group A (24 patients) underwent surgery because of symptomatic cervical myelopathy. Group B (43 patients) had no clinical symptoms but showed spinal cord compression on CTM. Group C (12 patients), who showed neither clinical symptoms nor cord changes, underwent CTM for lumbar lesion evaluation. AMF patterns were classified into 4 types according to their configurations on CTM (reversed T, Y, V, and O types). Results In group B, the reversed T type and Y type appeared significantly more often near the compressed portion (p<0.001). A similar tendency was seen in group A. The V and O types were most frequently observed in group C (p<0.001). Conclusion On CTM, contrast dye tends to flow into the AMF of the cervical cord when the spinal cord is compressed. We speculate that there may be 3 possible mechanisms for this phenomenon: deformation of the epipial layer of the AMF due to cervical cord compression, AMF dilatation due to atrophy of the anterior funiculus or anterior horn, and temporary AMF dilatation when it becomes an alternative route for cerebrospinal fluid circulation

Characterization of Moyamoya and Middle Cerebral Artery Diseases by Carotid Canal Diameter and RNF213 p.R4810K Genotype

[Objectives] It is sometimes difficult to differentiate middle cerebral artery disease from moyamoya disease because the two can present similarly yet have different treatment strategies. We investigated whether the presence of a narrow carotid canal and the RNF213 mutation can help differentiate between the two phenotypes. [Population and Methods] We analyzed 78 patients with moyamoya disease, 27 patients with middle cerebral artery disease, and 79 controls from 2 facilities. The carotid canal diameter was measured using computed tomography. The p.R4810K mutation was genotyped by TaqMan assay. A receiver operating characteristics analysis was performed to assess the significance of the carotid canal diameter for the accurate diagnosis of moyamoya disease. [Results] The carotid canal diameter was significantly narrower in patients with moyamoya disease than in controls. The optimal cutoff values were 5.0 mm for adult males and 4.5 mm for adult females and children (sensitivity: 0.82; specificity: 0.92). Among the patients with middle cerebral artery disease, 18.5% and 25.0% of the affected hemispheres had the p.R4810K mutation and narrow canal (i.e., below the cutoff), respectively, whereas only 3.1% of those had both. Contrastingly, 68.8% of the affected hemispheres in patients with moyamoya disease had both these characteristics. Among the patients with moyamoya disease, those with the p.R4810K mutation tended to have narrower carotid canals. [Conclusions] Although the presence of a narrow carotid canal or the p.R4810K mutation alone could not be used to distinguish those with moyamoya disease from those with middle cerebral artery disease, the combination of these factors could better characterize the two phenotypes