Phenotypic Drug Screening for Dysferlinopathy Using Patient-Derived Induced Pluripotent Stem Cells

Kokubu, Yuko; Nagino, Tomoko; Sasa, Katsunori; Oikawa, Tatsuo; Miyake, Katsuya; Kume, Akiko; Fukuda, Mikiko; Fuse, Hiromitsu; Tozawa, Ryuichi; Sakurai, Hiroyuki

doi:10.1002/sctm.18-0280

Cited by 34 publications

(43 citation statements)

References 28 publications

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“…Being a member of the ferlin family, dysferlin plays a key role in the active process of repairing muscle membrane lesions and can be widely expressed in a variety of tissues and cells, including skeletal and cardiac muscles, lung, kidney, brain, adipocytes and monocytes 23,24 . Being involved in T‐tubule formation and myogenesis, dysferlin contains a short carboxy‐terminal transmembrane domain and seven amino‐terminal C2 domains (C2A‐C2G) with a DYSF domain localized between C2C and C2D, among which the C2A domain binds to phospholipids in the form of Ca 2+ dependence 25–29 . Patients with LGMD2B may have deficient or absent dysferlin as a result of variants in the DYSF gene.…”

Section: Discussionmentioning

confidence: 99%

Compound heterozygous DYSF variants causing limb‐girdle muscular dystrophy type 2B in a Chinese family

Jing

Cheng

et al. 2020

The Journal of Gene Medicine

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Background: The dysferlin gene or the DYSF gene encodes the Ca 2+-dependent phospholipid-binding protein dysferlin, which belongs to the ferlin family and is associated with muscle membrane regeneration and repair. Variants in the DYSF gene are responsible for limb-girdle muscular dystrophy type 2B (LGMD2B), also called limb-girdle muscular dystrophy recessive 2 (LGMDR2), a rare subtype of muscular dystrophy involving progressive muscle weakness and atrophy. The present study aimed to identify the variants responsible for the clinical symptoms of a Chinese patient with limb girdle muscular dystrophies (LGMDs) and to explore the genotypephenotype associations of LGMD2B. Methods: A series of clinical examinations, including blood tests, magnetic resonance imaging scans for the lower legs, electromyography and muscle biopsy, was performed on the proband diagnosed with muscular dystrophies. Whole exome sequencing was conducted to detect the causative variants, followed by Sanger sequencing to validate these variants. Results: We identified two compound heterozygous variants in the DYSF gene, c.1058 T>C, p.(Leu353Pro) in exon 12 and c.1461C>A/p.Cys487* in exon 16 in this proband, which were inherited from the father and mother, respectively. In silico analysis for these variants revealed deleterious results by PolyPhen-2 (Polymorphism Phenotyping v2; http://genetics.bwh.harvard.edu/pph2), SIFT (Sorting Intolerant From Tolerant; https://sift.bii.a-star.edu.sg), PROVEAN (Protein Variation Effect Analyzer; http://provean.jcvi.org/seq_submit.php) and MutationTaster (http://www. mutationtaster.org). In addition, the two compound heterozygous variants in the proband were absent in 100 control individuals who had an identical ethnic origin and were from the same region, suggesting that these variants may be the pathogenic variants responsible for the LGMD2B phenotypes for this proband. Conclusions: The present study broadens our understanding of the mutational spectrum of the DYSF gene, which provides a deep insight into the pathogenesis of LGMDs and accelerates the development of a prenatal diagnosis.

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Section: Discussionmentioning

confidence: 99%

Compound heterozygous DYSF variants causing limb‐girdle muscular dystrophy type 2B in a Chinese family

Jing

Cheng

et al. 2020

The Journal of Gene Medicine

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“…New lipomuscular models of NNS would help in this regard. PSC-derived muscular disease models have contributed to the elucidation of pathophysiologies and drug discovery [19,35] and should be of benefit to understanding the pathophysiology of the muscular lesions in NNS. The in vivo effects of compounds on animal models should be also evaluated.…”

Section: Discussionmentioning

confidence: 99%

“…High-throughput compound screening (HTS) has been widely adopted as a strategy of drug discovery for intractable diseases including autoimmune disorders [16]. HTS combined with PSC-derived disease models has provided novel drug candidates for several diseases including amyotrophic lateral sclerosis, fibrodysplasia ossificans progressiva and dysferlinopathy [17][18][19]. Here we established an HTS system using the PSC-derived MLs as model of NNS (NNS-MLs) to discover effective therapeutic candidates for the disease.…”

Section: Introductionmentioning

confidence: 99%

High-throughput Screening with Pluripotent Stem Cells Identifies CUDC-907 as an Effective Compound for Restoring the Proinflammatory Phenotype of Nakajo-Nishimura Syndrome

Kase

Terashima

Ohta

et al. 2020

Preprint

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Nakajo-Nishimura syndrome (NNS) is an autoinflammatory disorder caused by a homozygous mutations in PSMB8 gene. The administration of systemic corticosteroids is partially effective, but continuous treatment causes severe side effects. We previously established a pluripotent stem cell (PSC)-derived NNS disease model that reproduces several inflammatory phenotypes including the overproduction of monocyte chemoattractant protein-1 (MCP-1) and interferon gamma-induced protein-10 (IP-10). Here we performed high-throughput compound screening (HTS) using this PSC-derived NNS model to find potential therapeutic candidates and identified CUDC-907 as an effective inhibitor of the release of MCP-1 and IP-10.CUDC-907 did not induce cell death within therapeutic concentrations and was also effective on primary patient cells. Further analysis indicated that the inhibitory effect was post-transcriptional. These findings suggest that HTS with PSC-derived disease models is useful for finding drug candidates for autoinflammatory diseases. Significance statementIn this study, we identified a histone deacetylase inhibitor CUDC-907 as a potential effective compound for ameliorating overproduction of inflammatory chemokines in an autoinflammatory disease named Nakajo-Nishimura syndrome. We performed high-throughput screening using pluripotent stem cell-derived monocytic cell lines. Our data prove the validity of screening system as a versatile platform for seeking candidate compounds for the treatment of congenital immunological disorders associated with monocytic lineage cells.

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“…Furthermore, these researchers demonstrated that it was possible to rescue the phenotype of MM by overexpressing DYSFERLIN using plasmid transfection [ 90 ]. Avoiding the use of genetic manipulation, Kokubu et al [ 91 ] developed a drug screening platform using iPSC-derived myocytes. They found that nocodazole was able to increase DYSFERLIN expression in cells.…”

Section: Disease Modelingmentioning

confidence: 99%

Versatility of Induced Pluripotent Stem Cells (iPSCs) for Improving the Knowledge on Musculoskeletal Diseases

Sanjurjo‐Rodríguez

Castro-Viñuelas

Piñeiro-Ramil

et al. 2020

IJMS

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Induced pluripotent stem cells (iPSCs) represent an unlimited source of pluripotent cells capable of differentiating into any cell type of the body. Several studies have demonstrated the valuable use of iPSCs as a tool for studying the molecular and cellular mechanisms underlying disorders affecting bone, cartilage and muscle, as well as their potential for tissue repair. Musculoskeletal diseases are one of the major causes of disability worldwide and impose an important socio-economic burden. To date there is neither cure nor proven approach for effectively treating most of these conditions and therefore new strategies involving the use of cells have been increasingly investigated in the recent years. Nevertheless, some limitations related to the safety and differentiation protocols among others remain, which humpers the translational application of these strategies. Nonetheless, the potential is indisputable and iPSCs are likely to be a source of different types of cells useful in the musculoskeletal field, for either disease modeling or regenerative medicine. In this review, we aim to illustrate the great potential of iPSCs by summarizing and discussing the in vitro tissue regeneration preclinical studies that have been carried out in the musculoskeletal field by using iPSCs.

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Phenotypic Drug Screening for Dysferlinopathy Using Patient-Derived Induced Pluripotent Stem Cells

Cited by 34 publications

References 28 publications

Compound heterozygous DYSF variants causing limb‐girdle muscular dystrophy type 2B in a Chinese family

Compound heterozygous DYSF variants causing limb‐girdle muscular dystrophy type 2B in a Chinese family

High-throughput Screening with Pluripotent Stem Cells Identifies CUDC-907 as an Effective Compound for Restoring the Proinflammatory Phenotype of Nakajo-Nishimura Syndrome

Versatility of Induced Pluripotent Stem Cells (iPSCs) for Improving the Knowledge on Musculoskeletal Diseases

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