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Towards next generation maggot debridement therapy: transgenic Lucilia sericata
larvae that produce and secrete a human growth factor
#MMPMID27006073
Linger RJ
; Belikoff EJ
; Yan Y
; Li F
; Wantuch HA
; Fitzsimons HL
; Scott MJ
BMC Biotechnol
2016[Mar]; 16
(?): 30
PMID27006073
show ga
BACKGROUND: Diabetes and its concurrent complications impact a significant
proportion of the population of the US and create a large financial burden on the
American health care system. FDA-approved maggot debridement therapy (MDT), the
application of sterile laboratory-reared Lucilia sericata (green bottle fly)
larvae to wounds, is a cost-effective and successful treatment for diabetic foot
ulcers and other medical conditions. Human platelet derived growth factor-BB
(PDGF-BB) is a secreted dimeric peptide growth factor that binds the PDGF
receptor. PDGF-BB stimulates cell proliferation and survival, promotes wound
healing, and has been investigated as a possible topical treatment for
non-healing wounds. Genetic engineering has allowed for expression and secretion
of human growth factors and other proteins in transgenic insects. Here, we
present a novel concept in MDT technology that combines the established benefits
of MDT with the power of genetic engineering to promote healing. The focus of
this study is to create and characterize strains of transgenic L. sericata that
express and secrete PDGF-BB at detectable levels in adult hemolymph, whole larval
lysate, and maggot excretions/ secretions (ES), with potential for clinical
utility in wound healing. RESULTS: We have engineered and confirmed transgene
insertion in several strains of L. sericata that express human PDGF-BB. Using a
heat-inducible promoter to control the pdgf-b gene, pdgf-b mRNA was detected via
semi-quantitative PCR upon heat shock. PDGF-BB protein was also detectable in
larval lysates and adult hemolymph but not larval ES. An alternative,
tetracycline-repressible pdgf-b system mediated expression of pdgf-b mRNA when
maggots were raised on diet that lacked tetracycline. Further, PDGF-BB protein
was readily detected in whole larval lysate as well as larval ES. CONCLUSIONS:
Here we show robust, inducible expression and production of human PDGF-BB protein
from two conditional expression systems in transgenic L. sericata larvae. The
tetracycline-repressible system appears to be the most promising as PDGF-BB
protein was detectable in larval ES following induction. Our system could
potentially be used to deliver a variety of growth factors and anti-microbial
peptides to the wound environment with the aim of enhancing wound healing,
thereby improving patient outcome in a cost-effective manner.
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