Modified Adenovirus Reduces De Novo Peritoneal Adhesions in Rats and Limits Off-Target Transfection. Role of EZH2 in Adhesion Formation.
OBJECTIVE
Adenovector encoding tissue plasminogen activator (tPA) was shown to reduce experimental peritoneal adhesion. We investigated the targeting potential of our modified adenovector, its ability to reduce adhesions and the epigenetic role of histone methyltransferase EZH2 in adhesion formation.


METHODS
Control lacZ, nonmodified tPA or modified tPA vectors were instilled in the peritoneal cavity after injury in de novo adhesions or after lysis of adhesions in recurrent adhesions. Adhesion severity was scored and adhesions and liver tissues were examined for adenovirus E4 gene and tPA mRNA expression. Levels of tPA, plasminogen activator inhibitor-1 (PAI-1), transforming growth factor-β1 (TGF-β1), and EZH2 expression were measured.


RESULTS
E4 transcripts were detected in adhesions of nonmodified and modified and in livers of nonmodified but not in livers of modified de novo adhesions. Both nonmodified (p = 0.021) and modified vectors (p = 0.036) reduced the severity of de novo adhesions compared to lacZ vector. Levels of tPA in nonmodified (p = 0.021) and modified adhesions (p = 0.001) were elevated while PAI-1 (p = 0.013 and p = 0.001, respectively) and TGF-β1 levels (p = 0.002 and p = 0.016, respectively) were reduced compared with lacZ group. All vectors were not expressed in recurrent adhesions and severity score were not different among groups. EZH2 levels were elevated in de novo nontreated (p = 0.001) and was further increased in recurrent (p = 0.001) nontreated adhesions compared with noninjured peritoneum.


CONCLUSIONS
Modified adenovirus successfully targeted de novo adhesions but not liver tissues and reduced the severity of de novo adhesions. EZH2 is involved in the development and progression of peritoneal adhesions.
a Department of Surgery , Faculty of Medicine, Minia University , El-Minia , Egypt.
Hussein M
HM
Atta
b Department of Obstetrics and Gynecology , Georgia Regents University , Augusta , Georgia , USA.
Ayman A
AA
Al-Hendy
c Department of Biochemistry , Faculty of Medicine, Minia University , El-Minia , Egypt.
Salama R
SR
Abdel Raheim
c Department of Biochemistry , Faculty of Medicine, Minia University , El-Minia , Egypt.
Hend
H
Abdel-Ghany
c Department of Biochemistry , Faculty of Medicine, Minia University , El-Minia , Egypt.
Khalid A
KA
Nasif
c Department of Biochemistry , Faculty of Medicine, Minia University , El-Minia , Egypt.
Ahlam M
AM
Abdellah
c Department of Biochemistry , Faculty of Medicine, Minia University , El-Minia , Egypt.
Nagwa M
NM
Zenhom
c Department of Biochemistry , Faculty of Medicine, Minia University , El-Minia , Egypt.
Heba S
HS
Kamel
D000256:Adenoviridae / Q000235:genetics*; D017504:Adenovirus E4 Proteins / Q000378:metabolism; D000818:Animals; D013505:Digestive System Surgical Procedures / Q000009:adverse effects; D004195:Disease Models, Animal; D000071221:Enhancer of Zeste Homolog 2 Protein / Q000235:genetics / Q000378:metabolism*; D044127:Epigenesis, Genetic; D005822:Genetic Vectors / Q000008:administration & dosage / Q000009:adverse effects*; D006801:Humans; D007322:Instillation, Drug; D008099:Liver / Q000473:pathology*; D008297:Male; D010532:Peritoneal Diseases / Q000209:etiology / Q000378:metabolism* / Q000473:pathology*; D017395:Plasminogen Activator Inhibitor 1 / Q000378:metabolism; D011183:Postoperative Complications / Q000209:etiology / Q000378:metabolism* / Q000473:pathology; D051381:Rats; D017208:Rats, Wistar; D000267:Tissue Adhesions / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D010959:Tissue Plasminogen Activator / Q000235:genetics / Q000378:metabolism; D014162:Transfection / Q000295:instrumentation / Q000379:methods; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
D017504:Adenovirus E4 Proteins; D017395:Plasminogen Activator Inhibitor 1; C547314:Serpine1 protein, rat; D053773:Transforming Growth Factor beta1; C578253:EZH2 protein, rat; D000071221:Enhancer of Zeste Homolog 2 Protein; D010959:Tissue Plasminogen Activator
gene therapy; histone methyltransferase; peritoneal adhesions; plasminogen activator inhibitor-1; tissue plasminogen activator; transforming growth factor beta-1