Remapping and mutation analysis of benign adult familial myoclonic epilepsy in a Japanese pedigree.

Benign adult familial myoclonic epilepsy (BAFME), alternatively named familial adult myoclonic epilepsy 1/familial cortical myoclonic tremor with epilepsy 1 (FAME1/FCMTE1), is a hereditary epileptic syndrome characterized by autosomal dominant inheritance, adult-onset tremulous hand movement, myoclonus, infrequent epileptic seizure and non-progressive course without cerebellar ataxia and dementia. We previously reported evidence for linkage of BAFME to the region between D8S1784 and D8S1694 on chromosome 8q. Subsequently, other research groups reported mapping of the same clinical syndrome to different chromosomal loci, 2p and 5p, in Italian (FAME2/FCMTE2) and French (FAME3/FCMTE3) families, respectively. In this study, we performed a genome-wide linkage analysis using 10K single-nucleotide polymorphism arrays and additional microsatellite markers to reconfirm the BAFME-linked region. The BAFME-linked region was mapped to 7.16 Mb spanned by rs1898287 and rs2891799 on chromosomes 8q23.3-8q24.13 with a maximum two-point logarithm of odds score of 6.0 for the marker rs1021897. Sequence analysis and copy-number variant analysis of all 38 genes localized in the candidate region were performed, but no pathogenic mutation was identified. We conclude that the etiology of BAFME remains to be solved, and further genetic studies, which may require analysis in non-coding regions of a gene, introns or intergenic spacer regions, are necessary to reveal its unknown mutations.

Adult-type metachromatic leukodystrophy with compound heterozygous ARSA mutations: a case report and phenotypic comparison with a previously reported case.

Metachromatic leukodystrophy (MLD) is an autosomal recessive lysosomal storage disease caused by a deficiency of arylsulfatase A. MLD is a heterogeneous disease with variable age at onset and variable clinical features. We evaluated a 33-year-old female patient who developed manifestations of disinhibitory behavior. She was diagnosed with MLD by genetic analysis, which revealed compound heterozygous ARSA missense mutations (p.G99D and p.T409I). The same combination of mutations was previously reported in a Japanese patient with similar symptoms. We performed additional, detailed neuropsychological tests with functional imaging on the current patient that demonstrated frontal lobe dysfunction. These results indicate that the mutations have important implications for genotype-phenotype correlation in MLD.

Sustained whole-body functional rescue in congestive heart failure and muscular dystrophy hamsters by systemic gene transfer.

BACKGROUND: The success of muscular dystrophy gene therapy requires widespread and stable gene delivery with minimal invasiveness. Here, we investigated the therapeutic effect of systemic delivery of adeno-associated virus (AAV) vectors carrying human delta-sarcoglycan (delta-SG) gene in TO-2 hamsters, a congestive heart failure and muscular dystrophy model with a delta-SG gene mutation. METHODS AND RESULTS: A single injection of double-stranded AAV serotype 8 vector carrying human delta-SG gene without the need of any physical or pharmaceutical interventions achieved nearly complete gene transfer and tissue-specific expression in the heart and skeletal muscles of the diseased hamsters. Broad and sustained (>12 months) restoration of the missing delta-SG gene in the TO-2 hamsters corrected muscle cell membrane leakiness throughout the body and normalized serum creatine kinase levels (a 50- to 100-fold drop). Histological examination revealed minimal or the absence of central nucleation, fibrosis, and calcification in the skeletal muscle and heart. Whole-body functional analysis such as treadmill running showed dramatic improvement, similar to the wild-type F1B hamsters. Furthermore, cardiac functional studies with echocardiography revealed significantly increased percent fractional shortening and decreased left ventricular end-diastolic and end-systolic dimensions in the treated TO-2 hamsters. The survival time of the animals was also dramatically extended. CONCLUSIONS: Systemic gene transfer of delta-SG by the AAV serotype 8 vector could effectively ameliorate cardiac and skeletal muscle pathology, profoundly improve cardiac and whole-body functions, and significantly prolong the lifespan of the treated TO-2 hamsters.

Differential expression of MMPs and TIMPs in moderate and severe heart failure in a transgenic model.

BACKGROUND: Altered expression of matrix metalloproteinases (MMPs) and their endogenous inhibitors (TIMPs) accompanies the development of heart failure (HF). However, changes in MMP and TIMP protein levels or activity during the progression from compensated to decompensated failure remains incompletely examined. METHODS AND RESULTS: Transgenic mice (Tg) with cardiac-specific overexpression of tumor necrosis factor-alpha (TNF1.6) develop a sex-related, progressive cardiac dilation and HF. Echocardiographic measures were used to categorize HF severity in male (M) and female (F) Tg and wild-type (WT) mice between 4 and 50 weeks of age. Cardiac TIMPs-1, TIMPs-2, and MMP-3 (enzyme-linked immunosorbent assay), and potential (APMA-activated) MMP-9 activity were measured at similar ages. In situ zymography assessed tissue gelatinase activity. Systolic function, ventricular dimensions, and presence of pleural effusions identified severe HF in younger M Tg mice (by 18 weeks) and older F Tg (>34 weeks). Regardless of age, sex, or HF severity, Tg mice expressed significantly more TIMP-1 (Tg 119-193 pg/mg vs. WT 13-24 pg/mg, P < .001) and potential MMP-9 activity (Tg 0.41-0.58 ng/mg vs. WT 0.015-0.028 ng/mg, P < .002). M Tg expressed elevated MMP-3 (4 weeks, 0.16 +/- 0.1 ng/mg protein vs. WT 0.04 +/- 0.01 ng/mg, P < .003), which increased with age and HF severity (18 weeks, 0.51 +/- 0.3 ng/mg P < .01). F Tg showed no increase in MMP-3 at 4 weeks but a progressive increase with age and HF severity (18 weeks 0.09 +/- 0.04 ng/mg, P < .02 vs. Tg M or WT; 34 weeks 0.13 +/- 0.02 ng/mg, P < .001 vs. WT). To test the hypothesis that increased MMP-3 may differentially activate MMP-9 in M Tg, in situ zymography was performed and revealed a significant increase in gelatinase activity in M Tg mice relative to both WT and F Tg. CONCLUSION: MMP-3 may regulate activation of MMP-9/gelatinase, the progression of cardiac remodeling, and development of decompensated heart failure.