PMC5870259_01
a Hematoxylin Eosin (HE) staining shows prominent perifascicular atrophy (asterisk). b HE staining highlights mononuclear infiltration around a blood vessel and infiltrating the endomysium. c CD163/CD4 immunostaining highlights several CD4 T-cells (red) in the perivascular and endomysial space with associated macrophages (brown) (d) MHC1 immunostaining shows abnormal sarcolemmal membrane overexpression in myocytes, note the normal control below should only highlight capillaries (e) C5b9 immunostaining highlights complement deposition in capillaries associated with affected fibers; note normal control does not show any capillary immunostain (f) Lamin A/C staining shows nuclear staining comparable to normal control. Scale bars: 50 mum
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Fig1
A muscle biopsy was obtained from her right quadriceps muscle which confirmed the earlier diagnosis of JDM, including perifascicular atrophy, perivascular mononuclear cell infiltrates and upregulation of sarcolemmal MHC1 (Fig. 1).
p.T10I LMNA mutation disrupts nuclear structure. a Integrated Genomics Viewer (IGV) screenshot of 130 bp paired-end whole exome sequencing reads, represented by horizontal gray bars, demonstrated a single nucleotide change (C to T) at genomic position of chromosome 1 g.156,084,738 (GRCh37/hg19) resulting in a missense alteration at codon 29 of LMNA replacing a threonine (T) with an isoleucine (I): NM_170707.3(LMNA):c.29C > T (p.T10I) (b) Skin fibroblast cell cultures of our patient with heterozygous p.T10I LMNA mutation and a Duchenne muscular dystrophy patient were analyzed by immunoflourescence using an antibody to lamin A. Cell cultures were stained with primary lamin A antibody, followed by 4',6-diamidino-2 phenylindole (DAPI) nuclear staining and with the Alexa 488 goat anti-rabbit IgG. Scale bar: 200 mum. c Higher magnification images of the nuclear staining shown in (b); scale bar: 50 mum. Arrows in panel (b) indicate fragmented nuclei; arrows in panel (c) indicate examples of atypically shaped nuclei and/or nuclei with ruffles of lamin A staining
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Fig2
T) in LMNA resulting in p.T10I substitution which is considered to be pathogenic (Fig. 2). Immunofluorescence staining of skin fibroblasts from the patient and a control patient with Duchenne muscular dystrophy which is used as a non- LMNA related control both displayed positive nuclear staining for lamin A (Fig. 2). Our results in Fig. 2 are consistent with previous reports that scored atypical nuclei which reported that control fibroblasts have occaisonal atypical nuclei in ~ 19% of cells while the number of atypical nuclei in fibroblasts from patients with lamin A/C mutations were much more common (33-92%).