In-vivo particle mediated delivery of mRNA to mammalian tissues: ballistic and biologic effects.
Biolistic transmission of mRNA provides transient gene therapy to in vivo organs. This study documents particle mediated mRNA transmission to a solid organ and wound healing model using the mRNA of Green Fluorescent Protein to determine optimal delivery parameters. Renal function, bullet penetration, cellular injury, and Green Fluorescent Protein synthesis were quantified. Chimeric human epidermal growth factor-FLAG epitope cDNA or mRNA was transmitted to wounds in normal or steroid treated animals. Wound bursting strength, human epidermal growth factor-FLAG, and collagen synthesis were determined. Injury and bullet penetration correlated with the delivery velocity and bullet size. Optimal delivery parameters were established which provided widespread Green Fluorescent Protein synthesis. Human epidermal growth factor-FLAG treatment significantly increased collagen content and wound breaking strength in normal and steroid treated animals. FLAG protein synthesis was evident in mRNA treated fascia following treatment. We found the gene gun provides a novel method for efficient, in vivo delivery of mRNA-based therapeutic strategies to mammalian organs with minimal histologic damage allowing transient expression of protein in in vivo target tissues. Co-delivery of Green Fluorescent Protein mRNA may provide a useful positive control to determine effective transmission. Biolistic transmission of human epidermal growth factor-FLAG mRNA provides increased tissue epidermal growth factor levels and accelerates wound healing in normal and steroid exposed animals.
Sohn
R L
RL
Department of Surgery, Wayne State University School of Medicine, Detroit, Michigan, USA.
Murray
M T
MT
Schwarz
K
K
Nyitray
J
J
Purray
P
P
Franko
A P
AP
Palmer
K C
KC
Diebel
L N
LN
Dulchavsky
S A
SA
D000818:Animals; D001682:Biological Availability; D004195:Disease Models, Animal; D016503:Drug Delivery Systems; D004815:Epidermal Growth Factor / Q000494:pharmacology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000502:physiology*; D008164:Luminescent Proteins; D008297:Male; D012333:RNA, Messenger / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012038:Regeneration; D012680:Sensitivity and Specificity; D014945:Wound Healing / Q000502:physiology; D014947:Wounds and Injuries / Q000235:genetics / Q000473:pathology / Q000628:therapy*
D008164:Luminescent Proteins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D004815:Epidermal Growth Factor