Matrix immobilization enhances the tissue repair activity of growth factor gene therapy vectors.
Although growth factor proteins display potent tissue repair activities, difficulty in sustaining localized therapeutic concentrations limits their therapeutic activity. We reasoned that enhanced histogenesis might be achieved by combining growth factor genes with biocompatible matrices capable of immobilizing vectors at delivery sites. When delivered to subcutaneously implanted sponges, a platelet-derived growth factor B-encoding adenovirus (AdPDGF-B) formulated in a collagen matrix enhanced granulation tissue deposition 3- to 4-fold (p < or = 0.0002), whereas vectors encoding fibroblast growth factor 2 or vascular endothelial growth factor promoted primarily angiogenic responses. By day 8 posttreatment of ischemic excisional wounds, collagen-formulated AdPDGF-B enhanced granulation tissue and epithelial areas up to 13- and 6-fold (p < 0.009), respectively, and wound closure up to 2-fold (p < 0.05). At longer times, complete healing without excessive scar formation was achieved. Collagen matrices were shown to retain both vector and transgene products within delivery sites, enabling the transduction and stimulation of infiltrating repair cells. Quantitative PCR and RT-PCR demonstrated both vector DNA and transgene mRNA within wound beds as late as 28 days posttreatment. By contrast, aqueous formulations allowed vector seepage from application sites, leading to PDGF-induced hyperplasia in surrounding tissues but not wound beds. Finally, repeated applications of PDGF-BB protein were required for neotissue induction approaching equivalence to a single application of collagen-immobilized AdPDGF-B, confirming the utility of this gene transfer approach. Overall, these studies demonstrate that immobilizing matrices enable the controlled delivery and activity of tissue promoting genes for the effective regeneration of injured tissues.
Doukas
J
J
Selective Genetics, San Diego, CA 92121, USA. jdoukas@selectivegenetics.com
Chandler
L A
LA
Gonzalez
A M
AM
Gu
D
D
Hoganson
D K
DK
Ma
C
C
Nguyen
T
T
Printz
M A
MA
Nesbit
M
M
Herlyn
M
M
Crombleholme
T M
TM
Aukerman
S L
SL
Sosnowski
B A
BA
Pierce
G F
GF
D000256:Adenoviridae / Q000235:genetics / Q000378:metabolism; D000818:Animals; D000077214:Becaplermin; D002921:Cicatrix / Q000139:chemically induced; D003094:Collagen / Q000378:metabolism*; D016503:Drug Delivery Systems / Q000009:adverse effects / Q000295:instrumentation / Q000379:methods*; D004423:Ear / Q000473:pathology; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D015316:Genetic Therapy / Q000009:adverse effects / Q000295:instrumentation / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics / Q000378:metabolism; D006099:Granuloma / Q000139:chemically induced; D006801:Humans; D006965:Hyperplasia / Q000139:chemically induced; D007150:Immunohistochemistry; D008297:Male; D009928:Organ Specificity; D010982:Platelet-Derived Growth Factor / Q000009:adverse effects / Q000235:genetics* / Q000378:metabolism / Q000627:therapeutic use; D019736:Prostheses and Implants; D020574:Proto-Oncogene Proteins c-sis / Q000009:adverse effects / Q000235:genetics / Q000378:metabolism / Q000627:therapeutic use; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D014161:Transduction, Genetic; D019076:Transgenes / Q000235:genetics; D014945:Wound Healing
D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D012333:RNA, Messenger; D000077214:Becaplermin; D003094:Collagen