Disease-modifying bioactivity of intravenous cardiosphere-derived cells and exosomes in mdx mice
Russell G. Rogers, Mario Fournier, Lizbeth Sanchez, Ahmed G. Ibrahim, Mark A. Aminzadeh, Michael I. Lewis, Eduardo Marbán
Russell G. Rogers, Mario Fournier, Lizbeth Sanchez, Ahmed G. Ibrahim, Mark A. Aminzadeh, Michael I. Lewis, Eduardo Marbán
View: Text | PDF | Corrigendum
Research Article Muscle biology Stem cells

Disease-modifying bioactivity of intravenous cardiosphere-derived cells and exosomes in mdx mice

Abstract

Dystrophin deficiency leads to progressive muscle degeneration in Duchenne muscular dystrophy (DMD) patients. No known cure exists, and standard care relies on the use of antiinflammatory steroids, which are associated with side effects that complicate long-term use. Here, we report that a single intravenous dose of clinical-stage cardiac stromal cells, called cardiosphere-derived cells (CDCs), improves the dystrophic phenotype in mdx mice. CDCs augment cardiac and skeletal muscle function, partially reverse established heart damage, and boost the regenerative capacity of skeletal muscle. We further demonstrate that CDCs work by secreting exosomes, which normalize gene expression at the transcriptome level, and alter cell signaling and biological processes in mdx hearts and skeletal muscle. The work reported here motivated the ongoing HOPE-2 clinical trial of systemic CDC delivery to DMD patients, and identifies exosomes as next-generation cell-free therapeutic candidates for DMD.

Authors

Russell G. Rogers, Mario Fournier, Lizbeth Sanchez, Ahmed G. Ibrahim, Mark A. Aminzadeh, Michael I. Lewis, Eduardo Marbán

×

Figure 3

Skeletal muscle improvements by systemic CDC or EXO delivery.

Options: View larger image (or click on image) Download as PowerPoint
Skeletal muscle improvements by systemic CDC or EXO delivery. (A) In vit...
(A) In vitro force-frequency relationship of WT, vehicle-, cardiosphere-derived cell–treated (CDC-treated), or CDC-derived exosome–treated (EXO-treated) mdx solei (n = 5–8 per group). Both CDC and EXO treatment shifted the force-frequency curve up and to the left. (B) Twitch and (C) tetanic force derived from A demonstrate that CDC and EXO treatment boosts the developed force by mdx solei (n = 5–8 per group). (D) Pooled data from E reveal less interstitial fibrosis in CDC- and EXO-treated mdx solei (n = 5–6 per group). (E) Representative Masson’s trichrome–stained micrographs from vehicle-, CDC-, and EXO-treated mdx solei. Scale bars: 100 μm. (F) Pooled data from E reveal more total myofibers in CDC- and EXO-treated mdx solei (n = 5–6 per group). (G) Transcriptome analysis of vehicle-, CDC-, and EXO-treated mdx solei using 2-dimensional hierarchical clustering of genes with at least 1.5-fold change between vehicle/WT, CDC/vehicle, and EXO/vehicle. Both CDC and EXO treatment reversed the transcriptomic profile and partially normalized gene expression. (H) Representative NF-κB immunoblot and quantification of NF-κB phosphorylation in mdx solei. CDC treatment normalized NF-κB phosphorylation (n = 5 per group). In contrast, NF-κB phosphorylation was preserved by EXO treatment (n = 5 per group). (I) Immunohistochemical staining for CD68 and α-sarcomeric actinin (α-SA) in vehicle-, CDC-, EXO-treated mdx solei. Scale bars: 50 μm. (J) Pooled data from I reveal a greater abundance of CD68+ pixels in CDC- and EXO-treated mdx solei (n = 6–7 per group). (K) Immunohistochemical staining for neonatal myosin heavy chain (nMHC) and laminin in vehicle-, CDC-, and EXO-treated mdx solei. Scale bars: 50 μm. (L) Pooled data from K reveal more nMHC+ myofibers in CDC- and EXO-treated mdx solei (n = 7 per group). Bar graphs depict mean ± SEM. Statistical significance was determined by ANOVA with P ≤ 0.05. When appropriate, a Newman-Keuls correction for multiple comparisons was applied. *Significantly different from WT; #significantly different from vehicle. Dashed lines in H represent splice sites among lanes run on the same immunoblot.