Large family cohorts of lymphoblastoid cells provide a new cellular model for investigating facioscapulohumeral muscular dystrophy

Jones, Takako I.; Himeda, Charis L.; Pérez, Daniel; Jones, Peter L.

doi:10.1016/j.nmd.2016.12.007

Cited by 21 publications

(34 citation statements)

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“…We identified 3 FSHD1 patients with gender and first degree relative matched controls, which displayed large reported differences in DUX4 expression from the LCL cohort described by Jones et al, 2017 33 . Namely: GSM16283 (FSHD1, 6RU, female, family 2) and matched control GSM16281; GSM16414 (FSHD1, 6RU, female, family 11) and matched control GSM16320; GSM16278 (FSHD1, 6RU, male, family 2) and matched control GSM16412.…”

Section: Resultsmentioning

confidence: 97%

“…Lymphocyte lines used were described in Jones et al, 2017 33 , where ethical permission is detailed. Primary FSHD and control myoblasts were described in Barro et al 2010 16 where ethical permission is detailed.…”

Section: Methodsmentioning

confidence: 99%

“…DUX4 can induce expression of immune system-related genes in myoblasts 20 , and inflammatory genes are dysregulated in FSHD muscle biopsies 21,32 . Recently a large library of 114 control and FSHD lymphoblastoid cell lines (LCLs) (EBV immortalised B-lymphocytes) from 12 FSHD1 affected families was published 33 . In contrast to studies of FSHD myoblasts all 61 FSHD LCLs displayed robust DUX4 expression, as well as DUX4 target genes (although only a few were evaluated).…”

Section: Introductionmentioning

confidence: 99%

“…In contrast to studies of FSHD myoblasts all 61 FSHD LCLs displayed robust DUX4 expression, as well as DUX4 target genes (although only a few were evaluated). Curiously, a small number of control LCLs also express DUX4, albeit at significantly lower levels to FSHD LCLs 33 . Of further relevance, an in frame DUX4-IGH fusion gene in which the homeodomains of DUX4 are fused to a clamp-like transactivation domain of IGH, presents in a significant subset of B-cell Acute Lymphoblastic Leukaemia (B-ALL).…”

Section: Introductionmentioning

confidence: 99%

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Lymphocytes contribute to DUX4 target genes in FSHD muscle biopsies

Banerji

Panamarova

Zammit

2019

Preprint

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dystrophy (FSHD) is an incurable myopathy linked to 22 overexpression of DUX4. However, DUX4 is difficult to detect in FSHD myoblasts and 23 target gene expression is not a consistent FSHD muscle biopsy biomarker, displaying 24 efficacy only on pathologically inflamed samples. Immune gene misregulation occurs in 25 FSHD muscle biopsies with DUX4 targets enriched for inflammatory processes. However, 26 assessment of the FSHD immune cell transcriptome, and the evaluation of DUX4 and target 27 gene expression has not yet been performed. We show that FSHD lymphoblastoid cell lines 28 (LCLs) display robust DUX4 expression, and express early and late DUX4 targets. 29 Moreover, genes elevated on FSHD LCLs are elevated in FSHD muscle biopsies, correlating 30 with DUX4 target activation and histological inflammation. These genes are importantly 31 unaltered in FSHD myoblasts/myotubes, implying a non-muscle source in biopsies. Our 32 results indicate an immune cell source of DUX4 and target gene expression in FSHD muscle 33 biopsies. 34 35 36Key Words: DUX4//FSHD/facioscapulohumeral muscular dystrophy/lymphocyte/skeletal 37 muscle 38 39 40Facioscapulohumeral muscular dystrophy (FSHD) is a prevalent (12/100,000 1 ) inherited 41 skeletal myopathy. Clinically, FSHD manifests as a descending skeletal muscle atrophy, 42 commencing in the facial muscles before progressing to the shoulder girdle, associating with 43 scapular winging, and latterly the muscles of the lower limb, resulting in foot drop 2,3 . The 44 pattern of muscle involvement in FSHD is characteristically left/right asymmetric 4 . 45Heterogeneity in pathology progression among first degree relatives, including monozygotic 46 twins, is also well described [5][6][7] . Moreover, systemic disruption in FSHD pathology is 47 suggested by several extra-muscular features including retinal telangiectasia similar to Coat's 48 disease 8-10 and sensorineural hearing loss 11,12 . 49 50 FSHD shows an autosomal dominant pattern of inheritance linked to hypomethylation of the 51 D4Z4 macro-satellite at chromosome 4q35 2 . This epigenetic modification can be achieved in 52 two ways: ~95% of cases (FSHD1 -MIM 158900) have truncation of the D4Z4 region to 1-53 10 repeats 13 , while the remaining ~5% (FSHD2 -MIM 158901) carry mutations in chromatin 54 modifying genes such as SMCHD1 14 , or more rarely, DNMT3B 15 . In addition to D4Z4 55 hypomethylation, FSHD patients also carry a permissive 4qA haplotype, encoding a poly(A) 56 signal 13 . Each 3.3kb D4Z4 repeat unit encodes an open reading frame for the transcription 57 factor double homeobox 4 (DUX4). Epigenetic derepression of D4Z4 via hypomethylation 58 permits expression of DUX4 transcripts from the most distal repeat, which are then stabilised 59 by the poly(A) signal for translation. Mis-expression of DUX4 protein is thus proposed to 60 underlie FSHD pathology. 61 62 How DUX4 drives pathology in FSHD is poorly understood. DUX4 expression in myoblasts 63 has been shown to induce a set of genes which are pro-apoptotic 16-18 ...

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Section: Resultsmentioning

confidence: 97%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Lymphocytes contribute to DUX4 target genes in FSHD muscle biopsies

Banerji

Panamarova

Zammit

2019

Preprint

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“…Total RNA was extracted from dissected mouse muscles homogenized in 10 volumes of TRIzol (ThermoFisher) using the TissueLyser LT (Qiagen), as per manufacturer's instructions followed by on-column DNaseI treatment and clean-up using the RNeasy mini kit (Qiagen). Quantitative DUX4-fl mRNA expression was analyzed using nested qRT-PCR, as described [41,50].…”

Section: Gene Expression Analysis By Rt-pcrmentioning

confidence: 99%

Transgenic mice expressing tunable levels of DUX4 develop characteristic facioscapulohumeral muscular dystrophy-like pathophysiology ranging in severity

Jones

Chew

Barraza-Flores

et al. 2018

Preprint

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AbstractBackgroundAll types of facioscapulohumeral muscular dystrophy (FSHD) are caused by the aberrant myogenic activation of the somatically silent DUX4 gene, which initiates a cascade of cellular events ultimately leading to FSHD pathophysiology. Therefore, FSHD is a dominant gain-of-function disease that is amenable to modeling by DUX4 overexpression. However, there is large variability in the patient population. Typically, progressive skeletal muscle weakness becomes noticeable in the second or third decade of life, yet there are many genetically FSHD individuals who develop symptoms much later in life or remain relatively asymptomatic throughout their lives. Conversely, in rare cases, FSHD may present clinically prior to 5-10 yrs of age, ultimately manifesting as a very severe early onset form of the disease. Thus, there is a need to control the timing and severity of pathology in FSHD-like models.MethodsWe have recently described a line of conditional DUX4 transgenic mice, FLExDUX4, that develop a myopathy upon induction of human DUX4-fl expression in skeletal muscle. Here, we use the FLExDUX4 mouse crossed with the skeletal muscle-specific and tamoxifen inducible line ACTAl-MerCreMer to generate a highly versatile bi-transgenic mouse model with chronic, low-level DUX4-fl expression and mild pathology, that can be induced to develop more severe FSHD-like pathology in a dose-dependent response to tamoxifen. We identified conditions to reproducibly generate models exhibiting mild, moderate, or severe DUX4-dependent pathophysiology, and characterized their progression.ResultsWe assayed DUX4-fl mRNA and protein levels, fitness, strength, global gene expression, histopathology, and immune response, all of which are consistent with an FSHD-like myopathic phenotype. Importantly, we identified sex-specific and muscle-specific differences that should be considered when using these models for preclinical studies.ConclusionsThe ACTA1-MCM;FLExDUX4 bi-transgenic mouse model expresses a chronic low level of DUX4-fl and has mild pathology and detectable muscle weakness. The onset and progression of moderate to severe pathology can be controlled via tamoxifen injection to provide consistent and readily screenable phenotypes for assessing therapies targeting DUX4-fl mRNA and protein. Thus, these FSHD-like mouse models can be used to study a range of DUX4-fl expression and pathology dependent upon investigator need, through controlled mosaic expression of DUX4.

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The evolution of DUX4 gene regulation and its implication for facioscapulohumeral muscular dystrophy

Jagannathan¹

2022

Biochimica et Biophysica Acta (BBA) - Molecular Basis of Diseas

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Large family cohorts of lymphoblastoid cells provide a new cellular model for investigating facioscapulohumeral muscular dystrophy

Cited by 21 publications

References 64 publications

Lymphocytes contribute to DUX4 target genes in FSHD muscle biopsies

Lymphocytes contribute to DUX4 target genes in FSHD muscle biopsies

Transgenic mice expressing tunable levels of DUX4 develop characteristic facioscapulohumeral muscular dystrophy-like pathophysiology ranging in severity

The evolution of DUX4 gene regulation and its implication for facioscapulohumeral muscular dystrophy

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