Various haploinsufficiency mechanisms in Pitt-Hopkins syndrome

Sparber, Peter; Filatova, Alexandra; Анисимова, И. В.; Маркова, Т Г; Voinova, Viktoria; Chuhrova, A L; Tabakov, Vyacheslav; Skoblov, Mikhail

doi:10.1016/j.ejmg.2020.104088

Cited by 8 publications

(8 citation statements)

References 14 publications

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“…For functional characterization of the c.1035–7A>G variant and due to the fact that SCN2A gene expression is brain restricted, a splicing minigene assay was performed. Exon 9, intron 9, and exon 10 with the adjusted intronic regions of the SCN2A gene were amplified from the proband genomic DNA and cloned into the pSpl3-Flu2 splicing vector ( Sparber et al, 2020 ). Wild-type (WT) plasmid and a plasmid carrying c.1035–7A>G variant (MUT) were separately transfected to HEK293T cells and 48 h post-transfection total RNA was extracted and RT-PCR was performed.…”

Section: Resultsmentioning

confidence: 99%

“…Exon 9, intron 9, and exon 10 with the adjusted intronic regions of the SCN2A gene were amplified from the proband genomic DNA. The PCR product was cloned into a pSpl3-Flu2 plasmid vector as previously described ( Sparber et al, 2020 ). Sanger sequencing was used for the selection of clones carrying wild-type (WT) and NG_008143.1 (NM_021007.2):c.1035–7A>G variant (MUT).…”

Section: Methodsmentioning

confidence: 99%

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Case Report: Phenotype-Driven Diagnosis of Atypical Dravet-Like Syndrome Caused by a Novel Splicing Variant in the SCN2A Gene

et al. 2022

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Febrile-associated epileptic encephalopathy is a large genetically heterogeneous group that is associated with pathogenic variants in SCN1A, PCDH19, SCN2A, SCN8A, and other genes. The disease onset ranges from neonatal or early-onset epileptic encephalopathy to late-onset epilepsy after 18 months. Some etiology-specific epileptic encephalopathies have target therapy which can serve as a clue for the correct genetic diagnosis. We present genetic, clinical, electroencephalographic, and behavioral features of a 4-year-old girl with epileptic encephalopathy related to a de novo intronic variant in the SCN2A gene. Initial NGS analysis revealed a frameshift variant in the KDM6A gene and a previously reported missense variant in SCN1A. Due to lack of typical clinical signs of Kabuki syndrome, we performed X-chromosome inactivation that revealed nearly complete skewed inactivation. Segregation analysis showed that the SCN1A variant was inherited from a healthy father. The proband had resistance to multiple antiseizure medications but responded well to sodium channel inhibitor Carbamazepine. Reanalysis of NGS data by a neurogeneticist revealed a previously uncharacterized heterozygous variant c.1035–7A>G in the SCN2A gene. Minigene assay showed that the c.1035–7A>G variant activates a cryptic intronic acceptor site which leads to 6-nucleotide extension of exon 9 (NP_066287.2:p.(Gly345_Gln346insTyrSer). SCN2A encephalopathy is a recognizable severe phenotype. Its electro-clinical and treatment response features can serve as a hallmark. In such a patient, reanalysis of genetic data is strongly recommended in case of negative or conflicting results of DNA analysis.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Case Report: Phenotype-Driven Diagnosis of Atypical Dravet-Like Syndrome Caused by a Novel Splicing Variant in the SCN2A Gene

et al. 2022

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“…Кальций-фосфат, напротив, высокой эффективности трансфекции не обеспечивал. Хотя другие клеточные линии хорошо переносят трансфекцию кальций- фосфатом [32], использованные нами линии миобластов по какой-то причине не производят эндоцитоз, необходимый для доставки siРНК таким способом. Кроме того, есть предположение, что, будучи молекулой РНК без оболочки из липопротеинов или подобных реагентов, siРНК в составе преципитатов с кальций-фосфатом быстро уничтожается внеклеточными РНК-нуклеазами.…”

Section: Discussionunclassified

Development of an approach to knockdown of DUX4 gene by siRNA in myoblasts derived from patients with FSHD

Krivosheeva¹,

Вяхирева²,

Табаков³

et al. 2021

Nauchno-Prakticheskii Zhurnal «Medicinskaia Genetika

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Миодистрофия Ландузи-Дежерина (МЛД) - распространённое наследственное заболевание, вызываемое эктопической экспрессией гена DUX4 в мышечных клетках. Метод подавления экспрессии генов при помощи siРНК зарекомендовал себя как эффективный и безопасный способ генной терапии без вмешательства непосредственно в геном. В данной работе мы продемонстрировали возможность трансфекции миобластов человека целевыми siРНК без ущерба жизнеспособности и дальнейшей дифференцировке этих клеток в миотубы. Также мы показали различия в профиле экспрессии генов-мишеней DUX4 между миобластами, полученными от здоровых людей и от пациентов с МЛД. В дальнейшем эти данные могут помочь при разработке эффективных и специфичных siРНК для терапии МЛД. Facioscapulohumeral dystrophy (FSHD) is a common hereditary disease caused by ectopic expression of the DUX4 gene in muscle cells. The method of suppressing gene expression using siRNA has been shown to be an effective and safe method of gene therapy without interfering directly with the genome. In this work, we demonstrated the possibility of transfecting human myoblasts with targeted siRNAs without compromising the viability and further differentiation of these cells into myotubes. We also showed differences in the expression profile of DUX4 target genes between healthy and patient-derived myoblasts. In the future, these data can be used to develop effective and specific siRNAs for the treatment of FSHD.

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“…Clinically, patients are characterized by intellectual disability, developmental delay, breathing abnormalities, absent or limited speech, motor delay, seizure, constipation, and facial features including wide mouth and a broad nasal base with a high bridge [5,[8][9][10][11]. Seizure activity is observed in approximately 30-50% of patients [4,5,12]. In addition, brain imaging studies iden-tify several abnormalities that vary between patients, including underdevelopment of the corpus callosum, smaller hippocampus, enlarged caudate nuclei, and cerebellar and vermis hypoplasia [9,10,[13][14][15].…”

Section: Tcf4 Mutations In Pitt-hopkins Syndromementioning

confidence: 99%

Molecular and Cellular Function of Transcription Factor 4 in Pitt-Hopkins Syndrome

2021

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Transcription factor 4 (TCF4, also known as ITF2 or E2-2) is a type I basic helix-loop-helix transcription factor. Autosomal dominant mutations in TCF4 cause Pitt-Hopkins syndrome (PTHS), a rare syndromic form of autism spectrum disorder. In this review, we provide an update on the progress regarding our understanding of TCF4 function at the molecular, cellular, physiological, and behavioral levels with a focus on phenotypes and therapeutic interventions. We examine upstream and downstream regulatory networks associated with TCF4 and discuss a range of in vitro and in vivo data with the aim of understanding emerging TCF4-specific mechanisms relevant for disease pathophysiology. In conclusion, we provide comments about exciting future avenues of research that may provide insights into potential new therapeutic targets for PTHS.

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Various haploinsufficiency mechanisms in Pitt-Hopkins syndrome

Cited by 8 publications

References 14 publications

Case Report: Phenotype-Driven Diagnosis of Atypical Dravet-Like Syndrome Caused by a Novel Splicing Variant in the SCN2A Gene

Case Report: Phenotype-Driven Diagnosis of Atypical Dravet-Like Syndrome Caused by a Novel Splicing Variant in the SCN2A Gene

Development of an approach to knockdown of DUX4 gene by siRNA in myoblasts derived from patients with FSHD

Molecular and Cellular Function of Transcription Factor 4 in Pitt-Hopkins Syndrome

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