Antagonism of the human epidermal growth factor receptor family controls disease severity in murine collagen-induced arthritis.
OBJECTIVE
To evaluate the therapeutic potential of the human epidermal growth factor receptor (HER) family inhibitor, herstatin, in an animal model of arthritis.


METHODS
Constructs of herstatin and modified tissue plasminogen activator (tPA)-herstatin were expressed in HEK 293T cells, and secreted protein was analyzed by Western blotting. Tissue PA-herstatin adenovirus (Ad-tPA-Her) was prepared, and titers established. Gene expression of Ad-tPA-Her was determined by polymerase chain reaction using HeLa cells. Pharmacokinetics of gene and protein expression in vivo in liver tissue and serum samples were confirmed via intravenous administration of Ad-tPA-Her. Clinical signs of disease were monitored in arthritic DBA/1 mice after therapeutic administration of Ad-tPA-Her, and histologic analysis of hind foot specimens was performed.


RESULTS
Native herstatin was not secreted in supernatants, while modified tPA-herstatin was detected in abundance. HeLa cells stably expressed the tPA-herstatin gene when infected with virus. Additionally, tPA-herstatin gene and protein expression was observed over time in mice treated with virus. Importantly, Ad-tPA-Her, when administered therapeutically to arthritic mice, controlled clinical and histologic signs of disease and reduced the number of joints with severe damage.


CONCLUSIONS
Our results support the notion that the human epidermal growth factor receptor family has a role in the progression of collagen-induced arthritis. The novel tPA-herstatin fusion protein could be used as an effective therapeutic tool for control of inflammatory disorders involving an angiogenic component.
Sumariwalla
Percy F
PF
Kennedy Institute of Rheumatology Division, Faculty of Medicine, Imperial College London, London, UK. p.sumariwalla@imperial.ac.uk
Jin
Pei
P
Zhang
Juan
J
Ni
Irene
I
Crawford
Damian
D
Shepard
H Michael
HM
Paleolog
Ewa M
EM
Feldmann
Marc
M
D000818:Animals; D001169:Arthritis, Experimental / Q000188:drug therapy / Q000473:pathology; D001172:Arthritis, Rheumatoid / Q000188:drug therapy* / Q000473:pathology; D006367:HeLa Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000096:biosynthesis* / Q000493:pharmacokinetics / Q000627:therapeutic use*; D008297:Male; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000493:pharmacokinetics / Q000627:therapeutic use*; D012720:Severity of Illness Index; D010959:Tissue Plasminogen Activator / Q000627:therapeutic use
D036341:Intercellular Signaling Peptides and Proteins; D011993:Recombinant Fusion Proteins; C434665:herstatin; C533324:tissue PA-herstatin adenovirus fusion protein; D010959:Tissue Plasminogen Activator