Genetic screening of Russian Usher syndrome patients toward selection for gene therapy

2018 ◽  
Vol 39 (6) ◽  
pp. 706-713 ◽  
Author(s):  
Marianna E. Ivanova ◽  
Vladimir N. Trubilin ◽  
Dmitry S. Atarshchikov ◽  
Andrey M. Demchinsky ◽  
Vladimir V. Strelnikov ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Genetic Screening ◽  
Usher Syndrome ◽  
Selection For

scholarly journals Usher syndrome gene mutation spectrum in Russian patients

2020 ◽  
pp. 38-39
Author(s):  
М.Е. Иванова ◽  
А.М. Демчинский ◽  
В.С. Каймонов ◽  
И.В. Миронова ◽  
И.В. Володин ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Retinitis Pigmentosa ◽  
Gene Mutation ◽  
Clinical Diagnosis ◽  
Usher Syndrome ◽  
Gene Mutations ◽  
Mutation Spectrum ◽  
Initial Sample ◽  
New Mutations

Изучение спектра мутаций и совершенствование диагностики синдрома Ашера (СА) особо актуальны в связи с разрабатываемыми подходами к генной терапии заболевания. Среди 46 пациентов с признаками СА патогенные мутации выявлены нами у 40 (87%) пациентов. СА I и II типов определены у 26% и 57% пробандов исходной выборки, соответственно. У пациентов с СА I выявлены мутации в генах MYO7A (73%), CDH23 (7%), PCDH15 (7%), и USH1C (13%). Наибольшую частоту показала мутация MYO7A p.Q18*. Описано 6 новых мутаций в гене MYO7A, и две - в гене PCDH15. У пациентов с СА II выявлена 21 мутация гена USH2A, 5 из которых описаны впервые. Наибольшую частоту показала мутация USH2A p.W3955*. У двух пациентов выявлены мутации в генах несиндромального пигментного ретинита RHO и RPGR, что позволило уточнить клинический диагноз. Studying the mutation spectrum and improvement of molecular verification of the Usher syndrome (USH) are of particular relevance as gene therapy emerges. Among 46 patients with signs of Usher syndrome we identified mutations in 40 (85%) patients, establishing a diagnosis of USH1 and USH2 for 26% and 57% of the probands of the initial sample, respectively. Patients with USH1 showed mutations in the MYO7A (73%), CDH23 (7%), PCDH15 (7%), and USH1C (13%) genes. MYO7A p.Q18* mutation showed the highest frequency. We have identified 6 new mutations in the MYO7A gene, and 2 in the PCDH15 gene. In USH2 patients, 21 USH2A gene mutations were identified, 5 of which are novel. The USH2A mutation p.W3955* was most frequent. Two patients showed mutations in the non-syndromic retinitis pigmentosa genes RHO and RPGR, which made it possible to clarify the clinical diagnosis.

scholarly journals Genetic Screening of the Usher Syndrome in Cuba

2019 ◽  
Vol 10 ◽  
Author(s):  
Elayne E. Santana ◽  
Carla Fuster-García ◽  
Elena Aller ◽  
Teresa Jaijo ◽  
Belén García-Bohórquez ◽  
...  
Keyword(s):  
Genetic Screening ◽  
Usher Syndrome

Limiting numbers of G156A O6-methylguanine–DNA methyltransferase-transduced marrow progenitors repopulate nonmyeloablated mice after drug selection

Blood ◽  
2000 ◽  
Vol 95 (10) ◽  
pp. 3078-3084 ◽  
Author(s):  
Brian M. Davis ◽  
Omer N. Koç ◽  
Stanton L. Gerson
Keyword(s):  
Gene Therapy ◽  
Dna Methyltransferase ◽  
Drug Induced ◽  
Methylguanine Dna Methyltransferase ◽  
Patients With Cancer ◽  
Colony Forming Units ◽  
Dominant Selectable Marker ◽  
Selection For

Abstract The limited efficacy of hematopoietic gene therapy can be improved by in vivo selection for transduced long-term repopulating cells (LTRC). We selected for G156A MGMT (▵MGMT) transduced LTRC present in 5 × 104 to 100 × 104 marrow cells infused into nonmyeloablated mice by the administration of O6-benzylguanine (BG) and BCNU every 3 to 4 weeks. To facilitate engraftment, mice were given a nonablative dose of BG and BCNU before infusion. Without selection, ▵MGMT was not detected in any hematopoietic colony-forming units (CFU) 24 to 30 weeks after infusion. After BG and BCNU, ▵MGMT+ CFU were frequently detected, and their proportions increased with each treatment cycle. After 2 to 3 cycles of BG and BCNU, many mice were stably reconstituted with 75% to 100% ▵MGMT+ CFU for at least 6 months, representing up to 940-fold enrichment. Thus, BG and BCNU stem cell toxicity allows ▵MGMT-transduced LTRC to repopulate the bone marrow. This degree of selection pressure in nonmyeloablated mice is far greater than that observed in previous drug-resistance gene transfer studies. These data support our approved clinical trial to select for drug-resistant, transduced hematopoietic cells, potentially decreasing cumulative drug-induced myelosuppression in patients with cancer. These data also suggest that ▵MGMT may be a potent, dominant, selectable marker for use in dual gene therapy.

scholarly journals Gene therapy approaches for prevention of retinal degeneration in Usher syndrome

Gene Therapy ◽  
2017 ◽  
Vol 24 (2) ◽  
pp. 68-71 ◽  
Author(s):  
D S Williams ◽  
A Chadha ◽  
R Hazim ◽  
D Gibbs
Keyword(s):  
Gene Therapy ◽  
Retinal Degeneration ◽  
Usher Syndrome

scholarly journals Gene Therapy Restores Balance and Auditory Functions in a Mouse Model of Usher Syndrome

2017 ◽  
Vol 25 (3) ◽  
pp. 780-791 ◽  
Author(s):  
Kevin Isgrig ◽  
Jack W. Shteamer ◽  
Inna A. Belyantseva ◽  
Meghan C. Drummond ◽  
Tracy S. Fitzgerald ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Mouse Model ◽  
Usher Syndrome ◽  
Auditory Functions

scholarly journals Local gene therapy durably restores vestibular function in a mouse model of Usher syndrome type 1G

2017 ◽  
Vol 114 (36) ◽  
pp. 9695-9700 ◽  
Author(s):  
Alice Emptoz ◽  
Vincent Michel ◽  
Andrea Lelli ◽  
Omar Akil ◽  
Jacques Boutet de Monvel ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Mouse Model ◽  
Inner Ear ◽  
Usher Syndrome ◽  
Vestibular Function ◽  
Gene Replacement ◽  
Syndrome Type ◽  
Mechanoelectrical Transduction ◽  
Efficient Gene ◽  
Usher Syndrome Type

Our understanding of the mechanisms underlying inherited forms of inner ear deficits has considerably improved during the past 20 y, but we are still far from curative treatments. We investigated gene replacement as a strategy for restoring inner ear functions in a mouse model of Usher syndrome type 1G, characterized by congenital profound deafness and balance disorders. These mice lack the scaffold protein sans, which is involved both in the morphogenesis of the stereociliary bundle, the sensory antenna of inner ear hair cells, and in the mechanoelectrical transduction process. We show that a single delivery of the sans cDNA by the adenoassociated virus 8 to the inner ear of newborn mutant mice reestablishes the expression and targeting of the protein to the tips of stereocilia. The therapeutic gene restores the architecture and mechanosensitivity of stereociliary bundles, improves hearing thresholds, and durably rescues these mice from the balance defects. Our results open up new perspectives for efficient gene therapy of cochlear and vestibular disorders by showing that even severe dysmorphogenesis of stereociliary bundles can be corrected.

scholarly journals Genetic screening revealed usher syndrome in a paediatric Chinese patient

2017 ◽  
Vol 15 (2) ◽  
pp. 98-106 ◽  
Author(s):  
Chunyan Qu ◽  
Fenghe Liang ◽  
Qin Long ◽  
Min Zhao ◽  
Haiqiong Shang ◽  
...  
Keyword(s):  
Genetic Screening ◽  
Usher Syndrome ◽  
Chinese Patient

scholarly journals Generation and Genetic Correction of USH2A c.2299delG Mutation in Patient-Derived Induced Pluripotent Stem Cells

Genes ◽  
2021 ◽  
Vol 12 (6) ◽  
pp. 805
Author(s):  
Xuezhong Liu ◽  
Justin Lillywhite ◽  
Wenliang Zhu ◽  
Zaohua Huang ◽  
Anna M. Clark ◽  
...  
Keyword(s):  
Stem Cells ◽  
Gene Therapy ◽  
Induced Pluripotent Stem Cells ◽  
Genome Editing ◽  
Pluripotent Stem Cells ◽  
Usher Syndrome ◽  
The United States ◽  
Specific Marker ◽  
Recessive Trait ◽  
Induced Pluripotent

Usher syndrome (USH) is the leading cause of inherited combined hearing and vision loss. As an autosomal recessive trait, it affects 15,000 people in the United States alone and is responsible for ~21% of inherited blindness and 3 to 6% of early childhood deafness. Approximately 2/3 of the patients with Usher syndrome suffer from USH2, of whom 85% have mutations in the USH2A gene. Patients affected by USH2 suffer from congenital bilateral progressive sensorineural hearing loss and retinitis pigmentosa which leads to progressive loss of vision. To study the molecular mechanisms of this disease and develop a gene therapy strategy, we generated human induced pluripotent stem cells (iPSCs) from peripheral blood mononuclear cells (PBMCs) obtained from a patient carrying compound heterozygous variants of USH2A c.2299delG and c.1256G>T and the patient’s healthy sibling. The pluripotency and stability were confirmed by pluripotency cell specific marker expression and molecular karyotyping. Subsequent CRISPR/Cas9 genome editing using a homology repair template was used to successfully correct the USH2A c.2299delG mutation back to normal c.2299G in the generated patient iPSCs to create an isogenic pair of lines. Importantly, this manuscript describes the first use of the recombinant Cas9 and synthetic gRNA ribonucleoprotein complex approach to correct the USH2A c.2299delG without additional genetic effects in patient-derived iPSCs, an approach that is amenable for therapeutic genome editing. This work lays a solid foundation for future ex vivo and in vivo gene therapy investigations and these patient’s iPSCs also provide an unlimited resource for disease modeling and mechanistic studies.

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