A novel dominant-negative mutation of the CSF1R gene causes adult-onset leukoencephalopathy with axonal spheroids and pigmented glia.

Adult-onset leukoencephalopathy with axonal spheroids and pigmented glia (ALSP) is a rare autosomal dominant disorder that is caused by mutations in the colony-stimulating factor 1 receptor (CSF1R) gene. Functional haplo-insufficiency of the CSF1R gene has been considered for the underlying genetic mechanisms. A novel mutation of CSF1R and its effects on CSF1R expression or clinical characteristics were explored in an ALSP family. Clinical data and imaging data were collected from the family members with ALSP. Peripheral blood samples were collected for DNA and RNA extraction. Whole-exome sequencing and quantitative PCR were used to identify mutations and to determine the expression of CSF1R. The family had a history of a dominant hereditary pattern. Patients in this family presented motor symptoms, emotional abnormality, or memory impairment at onset. MRI findings showed high hyperintensity signals of T2-weighted imaging in the white matter and atrophy of the corpus callosum. NOTCH3 gene sequencing ruled out the diagnosis of CADASIL. Whole-exome sequencing identified a novel splice-site mutation (c.2319+1C>A) in intron 16 of the CSF1R gene. CSF1R mRNA was significantly decreased (~15%) in the peripheral blood samples of affected patients, which was much lower than the expected 50%. Our findings not only supported the pathological implication of this splice-site mutation but also demonstrated for the first time a dominant-negative effect on CSF1R expression. This report extends the genetic spectrum of ALSP with CSF1R mutations and provides evidence for the clinical heterogeneity of ALSP.

A novel Alu-mediated microdeletion in the RUNX2 gene in a Chinese patient with cleidocranial dysplasia.

Cleidocranial dysplasia (CCD; OMIM: 119600) is a rare autosomal dominant skeletal dysplasia caused by RUNX2 gene mutations. The present study described a sporadic case with CCD. The clinical data of the proband with CCD was reported and genetic analysis was performed. The proband presented with typical CCD features including supernumerary impacted teeth, bilateral clavicle dysplasia, delayed closure of cranial sutures, and short stature; while his hands were normal. Sequencing analysis of the entire coding region of the RUNX2 gene revealed no pathogenic changes; however, copy-number analysis with the Affymetrix HD array found ~500 kb genomicmicrodeletion. Real-time quantitative PCR validated this microdeletion in the 1-4 exons of the RUNX2 gene. The junction point of the breaking DNA was located in the directly oriented AluSz6 and AluSx repetitive elements, indicating that this microdeletion might be generated through an Alu-Alu mediated mechanism. In addition, this microdeletion existed in 21.8% of the asymptomatic mother's peripheral blood cells, demonstrating that the mosaicism was not associated with CCD phenotypes. In summary, a pathogenic microdeletion in the RUNX2 gene located on chromosome 6 was responsible for CCD.

Serum cystatin C is associated with large cerebral artery stenosis in acute ischemic stroke.

Large cerebral artery stenosis is a major cause of acute ischemic stroke (AIS); however, the correlation between serum cystatin C (CysC) and the stenosis of large cerebral arteries in patients with AIS has not been established. We performed a retrospective review of acute ischemic stroke patients, who were examined by cerebral digital subtraction angiography(DSA). Participants (252 cases) included 131 patients without stenosis and 121 patients with large cerebral artery stenosis. Serum CysC levels in patients with large cerebral artery stenosis were much higher than that of control subjects (p<0.001). However, serum CysC levels were not related to the location of stenosis. Further, logistic regression analyses showed that increased serum CysC was an independent risk factor of large cerebral artery stenosis in patients with acute ischemic stroke. Total participants were subdivided into quintiles based on serum CysC levels. Compared with the first quintile, the odds ratios of risk for large cerebral artery stenosis in the fourth and the fifth quintile were 1.26 (p<0.05) and 4.71(p<0.05) respectively, after the adjustment for age, sex, and smoking, hypertension, type 2 diabetes mellitus(DM), dyslipidemia, creatinine(Cr), urea, uric acid, and C reactive protein(CRP). Therefore, a significant positive correlation was observed between elevated serum CysC levels and large cerebral artery stenosis in patients with acute ischemic stroke. In summary, our findings provide new insights into the correlation between increased serum CysC and large cerebral artery stenosis in patients with acute ischemic stroke.

A novel mutation of α-galactosidase A gene causes Fabry disease mimicking primary erythromelalgia in a Chinese family.

PURPOSE: Fabry disease is an X-linked genetic disorder caused by the mutations of α-galactosidase A (GLA, MIM 300644) gene presenting with various clinical symptoms including small-fiber peripheral neuropathy and limb burning pain. Here, we reported a Chinese pedigree with the initial diagnosis of primary erythromelalgia in an autosomal dominant (AD)-inherited pattern. METHODS: Mutation analysis of SCN9A and GLA genes by direct sequencing and functional analysis of a novel mutation of GLA in cells were performed. RESULTS: Our data did not show any pathological mutations in SCN9A gene; however, a novel missense mutation c.139T>C (p.W47R) of GLA was identified in a male proband as well as two female carriers in this family. Enzyme assay of α-galactosidase A activity showed deficient enzyme activity in male patients and female carriers, further confirming the diagnosis of Fabry disease. Finally, a functional analysis indicated that the replacement of the 47th amino acid tryptophan (W47) with arginine (W47R) or glycine (W47G) led to reduced activity of α-galactosidase A in 293T cells. Therefore, these findings demonstrated that the novel mutation p.W47R of GLA is the cause of Fabry disease. CONCLUSIONS: Because Fabry disease and primary erythromelalgia share similar symptoms, it is a good strategy for clinical physicians to perform genetic mutation screenings on both SCN9A and GLA genes in those patients with limb burning pain but without a clear inheritant pattern.

5-hydroxymethylcytosine is detected in RNA from mouse brain tissues.

5-hydroxymethylcytosine (5hmC) is considered as a novel DNA modification and plays an important role in cancer, stem cells, and developmental diseases. In this study, we demonstrated the existence of RNA 5hmC modification in mouse brain RNA by using a dot blot analysis method. Our data indicated that 5hmC modification in RNA samples was less than that in DNA samples. Further, we optimized the conditions for 5hmC detection in RNA samples such as DNase treatment, denature reagents, denature time, sample air-dry time, and the cross-linking time between RNA and membrane. Our results demonstrated that DNase treatment and denature reagents were two important factors that affected the 5hmC detection in RNA samples. By using the optimal conditions for RNA 5hmC detection, we found that the brainstem, the hippocampus, and the cerebellum had high levels of 5hmC modification and 5mC modification in RNA. Finally, we found that RNA 5hmC modification decreased in MPTP-induced Parkinson's disease model in mice. These suggest that 5hmC modification in RNA might play an important regulative role on protein or microRNA expression in these brain tissues. Because DNA 5hmC modification plays an important role in neural differentiation and development as well as neurological diseases, the significance of 5hmC modification in RNA in different neurological diseases needs further investigation. In summary, our study demonstrated for the first time the abundance of 5hmC modification in brain RNA by using a dot blot analysis method and proved that dot blot analysis is a useful method for 5hmC detection in RNA samples.