PROKR1 delivery by cell-derived vesicles restores the myogenic potential of Prokr1-deficient C2C12 myoblasts'''

Abstract

Cell-derived vesicles (CDVs) have been investigated as an alternative to exosomes. Here, we generated CDVs from Prokineticin receptor 1 (PROKR1) overexpressing HEK293T cells using micro-extrusion. More than 60 billion PROKR1-enriched CDV (PROKR1(Tg) CDVs) particles with canonical exosome properties were recovered from 10(7) cells. With 25 mu g/mL of PROKR1(Tg) CDVs, we observed delivery of PROKR1, significant reduction of apoptosis, and myotube formation in C2C12(Prokr1-/-) myoblasts that have lost their myogenic potential but underwent apoptosis following myogenic commitment. Expression levels of early and late myogenic marker genes and glucose uptake capacity were restored to equivalent levels with wild-type control. Furthermore, PROKR1(Tg) CDVs were accumulated in soleus muscle comparable to the liver without significant differences. Therefore, CDVs obtained from genetically engineered cells appear to be an effective method of PROKR1 protein delivery and offer promise as an alternative therapy for muscular dystrophy. (C) 2021 The Author(s). Published by Elsevier Inc.