[Developmental and epileptic encephalopathy produced by the ATP1A2 mutation]. / Entsefalopatiya s narusheniem razvitiya i epilepsiei, vyzvannaya mutatsiei gena ATP1A2.

A case of DEE98, a rare developmental and epileptic encephalopathy related to previously reported the de novo missense mutation p.Arg908Gln in the ATP1A2 gene, is described. A girl examined first time in 11 months had microcephaly, severe mental and motor delay, strabismus, spastic paraparesis and pachypolymicrogyria on brain MRI that is atypical for DEE98. Epilepsy with polymorphic seizures started at the age of 15 months. There was a remission lasting 9 months, after which seizures renewed. DEE98 was diagnosed at the age of 2 years 9 months by exome sequencing verified by trio Sanger sequencing. Another finding from high-throughput exome sequencing were two previously undescribed heterozygous variants of uncertain pathogenicity in the SPART gene, which causes autosomal recessive spastic paraplegia type 20 (SPG20); Sanger sequencing confirmed the trans position of the variants. The common clinical sign with typical SPG20 was early spastic paraparesis with contractures; other symptoms did not coincide. Considering the phenotypic diversity of SPG20 and the possibility of a combination of two independent diseases, we performed an additional study of the pathogenicity of SPART variants at the mRNA level: pathogenicity was not confirmed, and there were no grounds to diagnose SPG20.

The novel synonymous variant in LIPA gene affects splicing and causes lysosomal acid lipase deficiency.

Lysosomal acid lipase deficiency (LALD; MIM#278000) is a continuum of autosomal recessive diseases caused by defects in the gene LIPA and historically divided into two phenotypes: severe infantile-onset form called Wolman disease (WD) and childhood/adult-onset form known as cholesteryl ester storage disease (CESD). We report a novel synonymous homozygous variant c.600G > A in LIPA of a patient with LALD. Functional analysis of the patient cDNA and minigene assay revealed this variant as the cause of exonic cryptic splice site activation and 63 b.p. deletion in exon 6. To investigate the impact of this in-frame deletion on protein function, we performed 3D modeling of the human lysosomal acid lipase and showed the alteration of highly conservative region in close proximity to protein active site, which may completely eliminate the enzymatic activity. Using transcript specific real-time quantitative PCR method, we evaluated the relative ratio of the patient's wild type transcript isoform which is significantly reduced and correlates with severe childhood-onset variant of LALD.

Isolation and Characterization of Human Myoblast Culture In Vitro for Technologies of Cell and Gene Therapy of Skeletal Muscle Pathologies.

We analyzed cultures of 5 independent myoblast lines from human skeletal muscles. It was shown that the content of desmin-positive cells in cultures at early passages exceeds 90%. Typical morphofunctional signs of myogenic differentiation disturbances were identified and their dynamics was studied. Signs of alternative adipogenic and chondrogenic differentiation of cells were revealed. Based on these data, limitations for the use of myoblast cultures of certain passages for biomedical research and cell therapy were evaluated.

[Clinical and genetic characteristics of facioscapulohumeral muscular dystrophy Landuzi-Dezherina type 1]. / Kliniko-geneticheskie kharakteristiki litse-plechelopatochnoi miodistrofii Landuzi-Dezherina tipa 1.

AIM: To describe clinical and genetic characteristics of patients from the Russian population with a variety of phenotypic variants of facioscapulohumeral muscular dystrophy Landuzi-Dezherina type 1 (FSHD 1). MATERIAL AND METHODS: The material for the study were blood samples of 16 patients from 15 unrelated families residing in the territory of the Russian Federation, between the ages of 6 to 66 years, with symptoms of FSHD. Diagnosis was based on genealogical data analysis, neurological examination, electroneuromyographic study, indicators of activity of creatine phosphokinase (CPK) in the blood serum and molecular genetic analysis of the results, aimed at the analysis of macrosatellite D4Z4 repeats on chromosome 4. RESULTS AND CONCLUSION: The study established the diagnosis of FSHD1 in 75% of patients. The correlation of the severity and phenotypic spectrum of FSHD1 with the age of onset was found. There was the significant clinical heterogeneity even among the 1st degree relatives in the same family. The correlation between macrosatellite D4Z4 repeats and clinical features of FSHD1 described previously in the literature was not observed.

[The Mechanisms of Transgenerational Inheritance and Their Potential Contribution to Human Phenotypes].

As of today, classical genetics has already completed the majority of groundwork to describe the laws of inheritance, identify the causes of many human diseases, and dissect the mechanisms of transfer of genetic information from parents to offspring. However, recent studies indicate that inheritance of phenotypic traits may also occur through nongenetic factors, in particular, through epigenetic factors, that manifest their effects in a transgenerational fashion. This review discusses findings in the area of transgenerational inheritance that open a new era in modern genetics. We discuss the mechanisms of transgenerational inheritance, including DNA methylation, histone modifications, and noncoding RNA transfer, and give an overview of the approaches to detect transgenerational effects in humans.

[Cloning of alternative isoforms of the catalytic subunit of the human thelomerase (htert)].

Most human tumors, including cervical cancer, are characterized by telomerase activation (cell proliferation activation enzyme). Such activation is implemented in the elongation of the terminal segments (telomeres) of the telomerase chromosome. The gene of the enzyme is RNA-encoded, the RNA in tumors being observed in a few isoforms. The hTERT RNA role in cell activation and control was simulated using cervical cancer, as well as its pretumoral states (CIN), as a model object. The goal of this work was to clone of the human hTERT isoforms (normal, α-, ß-, and α+ß-splice-variants). The genetic constructions containing normal hTERT sequence, α- and ß-deletion variants based on the lentivirus vector pR780 were obtained. The α- and ß-deletion variants were not obtained in this variant because of methodological problems. In further research, we plan to implement splice-variants of hTERT in eukaryotic human cells.