scholarly journals Muscle pathophysiology in mouse models of musculocontractural Ehlers-Danlos syndrome due to CHST14 mutations (mcEDS-CHST14), generated through CRISPR/Cas9-mediated genomic editing

2021 ◽  
Author(s):  
Yuko Nitahara-Kasahara ◽  
Shuji Mizumoto ◽  
Yukiko U. Inoue ◽  
Shota Saka ◽  
Guillermo Posadas-Herrera ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Mouse Models ◽  
Genome Engineering ◽  
Dermatan Sulfate ◽  
Treatment Strategies ◽  
Mutant Mice ◽  
Wild Type ◽  
Ehlers Danlos Syndrome ◽  
Pathogenic Variants ◽  
Danlos Syndrome

Musculocontractural Ehlers-Danlos syndrome (mcEDS) is caused by generalized depletion of dermatan sulfate (DS) due to biallelic pathogenic variants in CHST14 encoding dermatan 4-O-sulfotransferase 1 (D4ST1) (mcEDS-CHST14). Here, we generated mouse models for mcEDS-CHST14 carrying homozygous mutations (1 bp deletion or 6 bp insertion/10 bp deletion) in Chst14 through CRISPR/Cas9-genome engineering to overcome perinatal lethality in conventional Chst14-deleted knockout mice. DS depletion was detected in the skeletal muscle of these genome-edited mutant mice, consistent with loss of D4ST1 activity. The mutant mice showed common pathophysiological features, regardless of the variant, including growth impairment and skin fragility. Notably, we identified myopathy-related phenotypes. Muscle histopathology showed variation in fiber size and spread of the muscle interstitium. Decorin localized diffusely in the spread endomysium and perimysium of skeletal muscle, unlike in wild-type mice. The mutant mice showed lower grip strength and decreased exercise capacity compared to wild-type, and morphometric evaluation demonstrated thoracic kyphosis in mutant mice. The established CRISPR/Cas9-engineered Chst14 mutant mice would be a useful model to further our understanding of the mcEDS pathophysiology and in the development of novel treatment strategies.

scholarly journals Myopathy Associated With Dermatan Sulfate-Deficient Decorin and Myostatin in Musculocontractural Ehlers-Danlos Syndrome: A Mouse Model Investigation

2021 ◽  
Vol 9 ◽  
Author(s):  
Yuko Nitahara-Kasahara ◽  
Guillermo Posadas-Herrera ◽  
Shuji Mizumoto ◽  
Aki Nakamura-Takahashi ◽  
Yukiko U. Inoue ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Dermatan Sulfate ◽  
Muscle Growth ◽  
Negative Regulator ◽  
Ehlers Danlos Syndrome ◽  
Pathogenic Variants ◽  
Myogenic Activity ◽  
Small Muscle ◽  
Danlos Syndrome ◽  
Deficient Mice

Carbohydrate sulfotransferase 14 (CHST14) encodes dermatan 4-O-sulfotransferase 1, a critical enzyme for dermatan sulfate (DS) biosynthesis. Musculocontractural Ehlers-Danlos syndrome (mcEDS) is associated with biallelic pathogenic variants of CHST14 and is characterized by malformations and manifestations related to progressive connective tissue fragility. We identified myopathy phenotypes in Chst14-deficient mice using an mcEDS model. Decorin is a proteoglycan harboring a single glycosaminoglycan chain containing mainly DS, which are replaced with chondroitin sulfate (CS) in mcEDS patients with CHST14 deficiency. We studied the function of decorin in the skeletal muscle of Chst14-deficient mice because decorin is important for collagen-fibril assembly and has a myokine role in promoting muscle growth. Although decorin was present in the muscle perimysium of wild-type (Chst14+/+) mice, decorin was distributed in the muscle perimysium as well as in the endomysium of Chst14–/– mice. Chst14–/– mice had small muscle fibers within the spread interstitium; however, histopathological findings indicated milder myopathy in Chst14–/– mice. Myostatin, a negative regulator of protein synthesis in the muscle, was upregulated in Chst14–/– mice. In the muscle of Chst14–/– mice, decorin was downregulated compared to that in Chst14+/+ mice. Chst14–/– mice showed altered cytokine/chemokine balance and increased fibrosis, suggesting low myogenic activity in DS-deficient muscle. Therefore, DS deficiency in mcEDS causes pathological localization and functional abnormalities of decorin, which causes disturbances in skeletal muscle myogenesis.

scholarly journals Delineation of musculocontractural Ehlers–Danlos Syndrome caused by dermatan sulfate epimerase deficiency

2020 ◽  
Vol 8 (5) ◽  
Author(s):  
Charlotte K. Lautrup ◽  
Keng W. Teik ◽  
Ai Unzaki ◽  
Shuji Mizumoto ◽  
Delfien Syx ◽  
...  
Keyword(s):  
Dermatan Sulfate ◽  
Ehlers Danlos Syndrome ◽  
Danlos Syndrome

scholarly journals Transcriptome Profiling of Primary Skin Fibroblasts Reveal Distinct Molecular Features Between PLOD1- and FKBP14-Kyphoscoliotic Ehlers–Danlos Syndrome

Genes ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 517 ◽  
Author(s):  
Lim ◽  
Lindert ◽  
Opitz ◽  
Hausser ◽  
Rohrbach ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Protein Trafficking ◽  
Treatment Strategies ◽  
Transcriptome Profiling ◽  
Joint Hypermobility ◽  
Skin Fibroblasts ◽  
Ehlers Danlos Syndrome ◽  
Molecular Features ◽  
Extracellular Matrix Components ◽  
Danlos Syndrome

Kyphoscoliotic Ehlers–Danlos Syndrome (kEDS) is a rare genetic heterogeneous disease clinically characterized by congenital muscle hypotonia, kyphoscoliosis, and joint hypermobility. kEDS is caused by biallelic pathogenic variants in either PLOD1 or FKBP14. PLOD1 encodes the lysyl hydroxylase 1 enzyme responsible for hydroxylating lysyl residues in the collagen helix, which undergo glycosylation and form crosslinks in the extracellular matrix thus contributing to collagen fibril strength. FKBP14 encodes a peptidyl-prolyl cis–trans isomerase that catalyzes collagen folding and acts as a chaperone for types III, VI, and X collagen. Despite genetic heterogeneity, affected patients with mutations in either PLOD1 or FKBP14 are clinically indistinguishable. We aim to better understand the pathomechanism of kEDS to characterize distinguishing and overlapping molecular features underlying PLOD1-kEDS and FKBP14-kEDS, and to identify novel molecular targets that may expand treatment strategies. Transcriptome profiling by RNA sequencing of patient-derived skin fibroblasts revealed differential expression of genes encoding extracellular matrix components that are unique between PLOD1-kEDS and FKBP14-kEDS. Furthermore, we identified genes involved in inner ear development, vascular remodeling, endoplasmic reticulum (ER) stress, and protein trafficking that were differentially expressed in patient fibroblasts compared to controls. Overall, our study presents the first transcriptomics data in kEDS revealing distinct molecular features between PLOD1-kEDS and FKBP14-kEDS, and serves as a tool to better understand the disease.

scholarly journals Duchenne muscular dystrophy patients lacking the dystrophin isoforms Dp140 and Dp71 and mouse models lacking Dp140 have a more severe motor phenotype

2021 ◽  
Author(s):  
Mary Chesshyre ◽  
Deborah Ridout ◽  
Yasumasa Hashimoto ◽  
Yoko Ookubo ◽  
Silvia Torelli ◽  
...  
Keyword(s):  
Skeletal Muscle ◽  
Duchenne Muscular Dystrophy ◽  
Muscular Dystrophy ◽  
Grip Strength ◽  
Mouse Models ◽  
Motor Function ◽  
Wild Type ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Background Duchenne muscular dystrophy (DMD) is caused by DMD mutations leading to dystrophin loss. Full length Dp427 is the primary dystrophin isoform expressed in skeletal muscle and is also expressed in the central nervous system (CNS). Two shorter isoforms, Dp140 and Dp71, are highly expressed in the CNS. While a role for Dp140 and Dp71 on DMD CNS co-morbidities is well known, relationships between lack of Dp140 and Dp71 and DMD motor outcomes are not. We have conducted a series of investigations addressing this. Methods Functional outcome data from 387 DMD boys aged 4.0-15.4 years was subdivided by DMD mutation expected effect on isoform expression; Group 1 (Dp427 absent, Dp140/Dp71 present, n=201); group 2 (Dp427/Dp140 absent, Dp71 present, n=152); and group 3 (Dp427/Dp140/Dp71 absent, n=34). Relationships between isoform group and North Star ambulatory assessment (NSAA) scores, 10m walk/run and rise times were explored using regression analysis. We used Capillary Western immunoassay (Wes) analysis to study Dp427, Dp140 and Dp71 production in wild-type and DMD skeletal muscle and myogenic cultures. Grip strength was studied in wild-type, mdx (Dp427 absent, Dp140/Dp71 present), mdx52 (Dp427/Dp140 absent, Dp71 present) and DMD-null (lacking all isoforms) mice. Results In DMD boys, we found a strong association between isoform group and motor function. In DMD boys, mean NSAA scores at 5 years of age were 6.1 points lower in group 3 than group 1 (p<0.01) and 4.9 points lower in group 3 than group 2 (p=0.05). Mean peak NSAA scores were 4.0 points lower in group 3 than group 1 (p<0.01), 2.4 points lower in group 3 than group 2 (p=0.09) and 1.6 points lower in group 2 than group 1 (p=0.04). Average grip strength in peak force at 3 months of age was higher in mdx than mdx52 mice (p=0.01). Dp427, but not Dp71, was produced in normal skeletal muscle; low levels of Dp71 were detected in DMD skeletal muscle. High Dp71 levels were present in wild-type and DMD myogenic cultures. Conclusions DMD boys lacking Dp140 and Dp140/Dp71 displayed worse motor function with a cumulative effect of isoform loss. DMD mouse models lacking Dp427 and Dp140 had lower grip strength than those lacking Dp427 but not Dp140. Our results highlight the importance of considering the effects of dystrophin isoform loss on DMD motor impairment, with important implications for understanding the complex relationship between brain and muscle function in DMD and patient stratification for clinical trials.

Genetic Heterogeneity and Clinical Variability in Musculocontractural Ehlers-Danlos Syndrome Caused by Impaired Dermatan Sulfate Biosynthesis

2015 ◽  
Vol 36 (5) ◽  
pp. 535-547 ◽  
Author(s):  
Delfien Syx ◽  
Tim Van Damme ◽  
Sofie Symoens ◽  
Merel C. Maiburg ◽  
Ingrid van de Laar ◽  
...  
Keyword(s):  
Genetic Heterogeneity ◽  
Dermatan Sulfate ◽  
Ehlers Danlos Syndrome ◽  
Clinical Variability ◽  
Danlos Syndrome

scholarly journals Recent Advances in the Pathophysiology of Musculocontractural Ehlers-Danlos Syndrome

Genes ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 43 ◽  
Author(s):  
Tomoki Kosho ◽  
Shuji Mizumoto ◽  
Takafumi Watanabe ◽  
Takahiro Yoshizawa ◽  
Noriko Miyake ◽  
...  
Keyword(s):  
Connective Tissue ◽  
Dermatan Sulfate ◽  
Close Contact ◽  
Normal Skin ◽  
Collagen Fibrils ◽  
Ehlers Danlos Syndrome ◽  
Reticular Dermis ◽  
Pathogenic Variants ◽  
Recent Advances ◽  
Skin Stretching

Musculocontractural Ehlers–Danlos Syndome (mcEDS) is a type of EDS caused by biallelic pathogenic variants in the gene for carbohydrate sulfotransferase 14/dermatan 4-O-sulfotransferase 1 (CHST14/D4ST1, mcEDS-CHST14), or in the gene for dermatan sulfate epimerase (DSE, mcEDS-DSE). Thus far, 41 patients from 28 families with mcEDS-CHST14 and five patients from four families with mcEDS-DSE have been described in the literature. Clinical features comprise multisystem congenital malformations and progressive connective tissue fragility-related manifestations. This review outlines recent advances in understanding the pathophysiology of mcEDS. Pathogenic variants in CHST14 or DSE lead to reduced activities of relevant enzymes, resulting in a negligible amount of dermatan sulfate (DS) and an excessive amount of chondroitin sulfate. Connective tissue fragility is presumably attributable to a compositional change in the glycosaminoglycan chains of decorin, a major DS-proteoglycan in the skin that contributes to collagen fibril assembly. Collagen fibrils in affected skin are dispersed in the papillary to reticular dermis, whereas those in normal skin are regularly and tightly assembled. Glycosaminoglycan chains are linear in affected skin, stretching from the outer surface of collagen fibrils to adjacent fibrils; glycosaminoglycan chains are curved in normal skin, maintaining close contact with attached collagen fibrils. Homozygous (Chst14−/−) mice have been shown perinatal lethality, shorter fetal length and vessel-related placental abnormalities. Milder phenotypes in mcEDS-DSE might be related to a smaller fraction of decorin DS, potentially through residual DSE activity or compensation by DSE2 activity. These findings suggest critical roles of DS and DS-proteoglycans in the multisystem development and maintenance of connective tissues, and provide fundamental evidence to support future etiology-based therapies.

scholarly journals Gene Defect of Dermatan Sulfate Proteoglycan of Cattle Affected With a Variant Form of Ehlers-Danlos Syndrome

1999 ◽  
Vol 13 (3) ◽  
pp. 202-205 ◽  
Author(s):  
Motoshi Tajima ◽  
Sachiko Miyake ◽  
Kazushige Takehana ◽  
Ataru Kobayashi ◽  
Osamu Yamato ◽  
...  
Keyword(s):  
Dermatan Sulfate ◽  
Variant Form ◽  
Sulfate Proteoglycan ◽  
Gene Defect ◽  
Ehlers Danlos Syndrome ◽  
Dermatan Sulfate Proteoglycan ◽  
Danlos Syndrome

scholarly journals Biochemical and thermodynamic characterization of mutated β1,4-galactosyltransferase 7 involved in the progeroid form of the Ehlers–Danlos syndrome

2010 ◽  
Vol 432 (2) ◽  
pp. 303-311 ◽  
Author(s):  
Sophie Rahuel-Clermont ◽  
Franck Daligault ◽  
Marie-Helene Piet ◽  
Sandrine Gulberti ◽  
Patrick Netter ◽  
...  
Keyword(s):  
Molecular Basis ◽  
Ex Vivo ◽  
Soluble Proteins ◽  
Titration Calorimetry ◽  
Wild Type ◽  
Gene Encoding ◽  
Ehlers Danlos Syndrome ◽  
Chain Initiation ◽  
Danlos Syndrome

Three mutations of the B4GALT7 gene [encoding β1,4-GalT7 (β1,4-galactosyltransferase 7)], corresponding to A186D, L206P and R270C, have been identified in patients with the progeroid form of the Ehlers–Danlos syndrome and are described as being associated with the reduction or loss of β1,4-GalT7 activity. However, the molecular basis of the reduction or loss of activity remained to be determined. In the present study, wild-type, A186D, L206P and R270C β1,4-GalT7 were expressed in CHO618 cells as membrane proteins and in Escherichia coli as soluble proteins fused to MBP (maltose-binding protein). The ability of the expressed proteins to transfer galactose from donor to acceptor substrates was systematically characterized by kinetic analysis. The physicochemical properties of soluble proteins were explored by isothermal titration calorimetry, which is a method of choice when determining the thermodynamic parameters of the binding of substrates. Together, the results showed that: (i) the L206P mutation abolished the activity when L206P β1,4GalT7 was either inserted in the membrane or expressed as a soluble MBP–full-length fusion protein; (ii) the A186D mutation weakly impaired the binding of the donor substrate; and (iii) the R270C mutation strongly impaired the binding of the acceptor substrate. Moreover, the ex vivo consequences of the mutations were investigated by evaluating the priming efficiency of xylosides on GAG (glycosaminoglycan) chain initiation. The results demonstrate a quantitative effect on GAG biosynthesis, depending on the mutation; GAG biosynthesis was fully inhibited by the L206P mutation and decreased by the R270C mutation, whereas the A186D mutation did not affect GAG biosynthesis severely.

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