CD20-related signaling pathway is differently activated in normal and dystrophic circulating CD133+ stem cells

Abstract

Among the heterogeneous population of circulating hematopoietic and endothelial progenitors, we identified a subpopulation of CD133(+) cells displaying myogenic properties. Unexpectedly, we observed the expression of the B-cell marker CD20 in blood-derived CD133(+) stem cells. The CD20 antigen plays a role in the modulation of intracellular calcium homeostasis through signaling pathways activation. Several observations suggest that an increase in intracellular calcium concentration ([Ca(2+)](i)) could be involved in the etiology of the Duchenne muscular dystrophy (DMD). Here, we show that a CD20-related signaling pathway able to induce an increase in [Ca(2+)](i) is differently activated after brain derived neurotrophic factor (BDNF) stimulation of normal and dystrophic blood-derived CD133(+) stem cells, supporting the assumption of a "CD20-related calcium impairment" affecting dystrophic cells. Presented findings represent the starting point toward the expansion of knowledge on pathways involved in the pathology of DMD and in the behavior of dystrophic blood-derived CD133(+) stem cells.

Defective dystrophic thymus determines degenerative changes in skeletal muscle

“…Total proteins from skeletal muscles isolated from transplanted nude mice (nu PBS n = 4; Tnu MDX n = 4; Tnu C57BL n = 4) were obtained as in ref. 84 . Samples were resolved on polyacrylamide gels (ranging from 6 to 10%) and transferred to nitrocellulose membranes (Bio-Rad Laboratories, CA, USA).…”

Therapeutic Potential of Immunoproteasome Inhibition in Duchenne Muscular Dystrophy

“…WB analysis. Total protein concentration was obtained as previously described 61 from muscle tissue samples of treated and untreated mdx mice, and from freshly isolated and IFN-γ supplemented myoblasts-isolated from C57Bl mice. Samples were resolved on an 6% polyacrylamide gel, transferred to supported nitrocellulose membranes (Bio-Rad Laboratories, Berkeley, CA) and the filters incubated overnight at 4°C with the following primary antibodies: anti-PSMB5 (1:500; Abcam, Cambridge, Great Britain); anti-PSMB8 (1:500; Abcam, Cambridge, UK); anti-PSMB9 (1:500; Abcam); anti-β tubulin III (1:400; Sigma, Saint-Louis); anti-GAPDH (1:1,000; Santa Cruz, Dallas, TX); anti-IL1β (1:1,000; Abcam-ab9722); antidystrophin (1:50; Novocastra); anti-β-sarcoglycan (1:500; Santa Cruz).…”

Fibrosis Rescue Improves Cardiac Function in Dystrophin-Deficient Mice and Duchenne Patient–Specific Cardiomyocytes by Immunoproteasome Modulation

“…Hearts were isolated from ONX-0914etreated and untreated mdx mice and total protein concentration obtained as previously described. 25 Samples were resolved on polyacrylamide gels (ranging from 6% to 12%) and transferred to nitrocellulose membranes (Bio-Rad Laboratories, Hercules, CA). The membranes were incubated overnight with the primary antibodies against: PSMB5 (1:500; Abcam); PSMB8 (1:500; Abcam); PSMB9 (1:500; Abcam); b tubulin III (1:400; Sigma-Aldrich); vinculin (1:1000; Santa Cruz Biotechnology, Santa Cruz, TX); osteopontin (1:500; R&D Systems, Minneapolis, MN); FoxP3 (1:500; eBioscience, San Diego, CA); TNF-a (1:500; eBioscience); transient receptor potential channel 1 (TRPC1; 1:500; Santa Cruz Biotechnology); STAT1 (1:400; Cell Signaling Technology); phosphorylated STAT1 (1:400; Cell Signaling Technology); STAT-3 (1:400; Cell Signaling Technology); and phosphorylated STAT3 (1:400; Cell Signaling Technology).…”