Simplified production of a recombinant human angiostatin derivative that suppresses intracerebral glial tumor growth.
Angiostatin is an endogenous inhibitor of tumor neovascularization that inhibits the proliferation of endothelial cells. Production of sufficient quantities of biologically active angiostatin by the enzymatic cleavage of plasminogen has proven difficult in that it has delayed clinical testing. We have cloned, expressed, and purified a recombinant human angiostatin derivative (K1-3) using a mammalian expression system. Through the addition of a secretory signal and polyhistidine sequence tag, K1-3 can be purified from post-culture medium by simple column chromatography. Purified K1-3 protein is apparently folded in an active conformation, as evidenced by its ability to bind to lysine-Sepharose. In vitro, recombinant K1-3 significantly suppressed endothelial cell proliferation in a dose-dependent manner with an IC50 of 50 nM. Using an animal model of intracranial brain tumors in immune-competent rats, systemic administration of purified recombinant K1-3 resulted in up to 85% suppression of tumor growth (P = 0.011). Growth suppression was accompanied by a 32% decrease (P = 0.01) in tumor neovascularization. This study demonstrates a simple method to produce a biologically active recombinant angiostatin derivative. The ability to suppress intracerebral tumor growth after systemic administration suggests that K1-3 is likely to have therapeutic value in the treatment of malignant glial tumors.
Meneses
P I
PI
Department of Neurosciences, Weill Medical College of Cornell University, New York, New York 10021, USA.
Abrey
L E
LE
Hajjar
K A
KA
Gultekin
S H
SH
Duvoisin
R M
RM
Berns
K I
KI
Rosenfeld
M R
MR
D000818:Animals; D001932:Brain Neoplasms / Q000098:blood supply / Q000188:drug therapy* / Q000473:pathology; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D005260:Female; D005910:Glioma / Q000098:blood supply / Q000188:drug therapy* / Q000473:pathology; D006801:Humans; D008833:Microcirculation / Q000187:drug effects / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000517:prevention & control*; D010446:Peptide Fragments / Q000235:genetics / Q000627:therapeutic use* / Q000633:toxicity; D010958:Plasminogen / Q000235:genetics / Q000627:therapeutic use* / Q000633:toxicity; D051381:Rats; D011916:Rats, Inbred F344; D011994:Recombinant Proteins / Q000627:therapeutic use / Q000633:toxicity; D014162:Transfection; D014471:Umbilical Veins
D010446:Peptide Fragments; D011994:Recombinant Proteins; C402400:recombinant kringle 1-3 (rKI-3) of human plasminogen; D010958:Plasminogen