Prevention of radiation-induced oral mucositis after adenoviral vector-mediated transfer of the keratinocyte growth factor cDNA to mouse submandibular glands.
OBJECTIVE
The study aims to evaluate if human keratinocyte growth factor (hKGF), secreted after transduction of murine salivary glands with adenoviral vectors, can prevent oral mucositis resulting from radiation.


METHODS
Two serotype 5 adenoviral vectors encoding hKGF were constructed: AdEF1alpha-hKGF and AdLTR(2)EF1alpha-hKGF. Female C3H mice, 8 weeks old, were irradiated by single (22.5 Gy) or fractionated (5 x 8 Gy for 5 days) doses to induce oral mucositis (ulcers on tongue). One day before irradiation, the above viral vectors or an empty vector, Adcontrol, was given (10(10) particles per gland) to both submandibular glands by retrograde ductal instillation. Each experiment included five groups: no irradiation and irradiation (+/-Adcontrol, AdEF1alpha-hKGF, or AdLTR(2)EF1alpha-hKGF). Blood, saliva, submandibular glands, and tongue were collected on day 7 for single-dose studies or day 10 for fractionated dosing. hKGF levels were measured by ELISA.


RESULTS
In three separate single-dose irradiation experiments, lingual ulcers were dramatically reduced after either KGF-expressing vector. Similarly, in two separate fractionated irradiation experiments, the hKGF-expressing vectors completely prevented ulcer formation. QPCR data indicated that approximately 10(7) to 10(8) particles of each vector remained in the targeted submandibular glands at the terminal time. Transgenic hKGF protein was found at high levels in saliva, serum, and submandibular gland extracts.


CONCLUSIONS
hKGF gene transfer to salivary glands prevented radiation-induced oral mucositis in mice. This proof of concept study suggests that transgenic hKGF secreted from transduced salivary glands may be useful clinically to prevent oral mucositis caused by radiation.
Zheng
Changyu
C
Molecular Physiology and Therapeutics Branch, National Institute of Dental and Craniofacial Research, National Cancer Institute, Bethesda, Maryland, USA.
Cotrim
Ana P
AP
Sunshine
Abraham N
AN
Sugito
Takayuki
T
Liu
Lina
L
Sowers
Anastasia
A
Mitchell
James B
JB
Baum
Bruce J
BJ
D000256:Adenoviridae / Q000235:genetics; D000704:Analysis of Variance; D000818:Animals; D002460:Cell Line; D018076:DNA, Complementary / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D051523:Fibroblast Growth Factor 7 / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D051379:Mice; D008809:Mice, Inbred C3H; D011833:Radiation Injuries, Experimental / Q000150:complications / Q000517:prevention & control*; D012463:Saliva / Q000737:chemistry; D013280:Stomatitis / Q000209:etiology / Q000473:pathology / Q000517:prevention & control*; D013363:Submandibular Gland / Q000378:metabolism / Q000473:pathology / Q000528:radiation effects*; D014161:Transduction, Genetic
D018076:DNA, Complementary; C496453:FGF7 protein, human; D051523:Fibroblast Growth Factor 7