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

Chunjuan Zhang; Jongsoo Mok; Yeonwoo Seong; Hui-Chong Lau; Dayeon Kim; Junsik Yoon; Seung Wook Oh; Tae Sub Park; Joonghoon Park

doi:10.1016/j.nano.2021.102448

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

Nanomedicine. 2021 Oct:37:102448. doi: 10.1016/j.nano.2021.102448. Epub 2021 Jul 25.

Authors

Chunjuan Zhang¹, Jongsoo Mok², Yeonwoo Seong¹, Hui-Chong Lau³, Dayeon Kim³, Junsik Yoon³, Seung Wook Oh³, Tae Sub Park⁴, Joonghoon Park⁵

Affiliations

¹ Department of International Agricultural Technology, Graduate School of International Agricultural Technology, Seoul National University, Republic of Korea.
² Institute of GreenBio Science and Technology, Seoul National University, Republic of Korea.
³ Biodrone Research Institute, MDimune Inc., Republic of Korea.
⁴ Department of International Agricultural Technology, Graduate School of International Agricultural Technology, Seoul National University, Republic of Korea; Institute of GreenBio Science and Technology, Seoul National University, Republic of Korea.
⁵ Department of International Agricultural Technology, Graduate School of International Agricultural Technology, Seoul National University, Republic of Korea; Institute of GreenBio Science and Technology, Seoul National University, Republic of Korea. Electronic address: [email protected].

PMID: 34314870
DOI: 10.1016/j.nano.2021.102448

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 10⁷ cells. With 25 μg/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.

Keywords: C2C12; Cell-derived vesicles; Muscular dystrophy; Myogenesis; PROKR1.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / drug effects*
Cell Differentiation / drug effects
Cell-Derived Microparticles / chemistry*
HEK293 Cells
Humans
Mice
Muscle Development / drug effects*
Muscle Development / genetics
Muscle Fibers, Skeletal / drug effects
Myoblasts / drug effects
Receptors, G-Protein-Coupled / chemistry*
Receptors, G-Protein-Coupled / genetics

Substances

PKR1 protein, mouse
Receptors, G-Protein-Coupled