Construction of an EGF receptor-mediated histone H1(0)-based gene delivery system.
OBJECTIVE
To construct an EGF receptor (EGF-R)-mediated histone H1(0)-based gene delivery system for gene therapy.


METHODS
A recombinant DNA containing histone H1(0), EGF-R ligand, and endosomalytic domains was constructed in a prokaryotic vector and expressed in E. coli. Expression of the beta-galactosidase (beta-gal) gene in the tumor cells and tissues was observed after transduction of the beta-gal gene packaged by purified fusion proteins in vitro and in vivo.


RESULTS
As an extension of the research on previously reported chemically synthetic composite polypeptide gene delivery systems, this genetically engineered polypeptide has proved to be capable of targeting the beta-galactosidase (beta-gal) gene into EGF-R-positive cancer cells both in vitro and in vivo. We also studied the time course of beta-gal gene expression in tumor tissues delivered in vivo by this polypeptide vector. At 24 h after administration, expression of the beta-galactosidase gene in tumor reached peak levels. The dosage optimization of administered polyplex was also investigated. The optimal dose of polyplex per mouse was 1 microg DNA packaged by 3 microg of composite polypeptide.


CONCLUSIONS
The genetically engineered polypeptide based on histone H1(0) is a promising gene delivery system targeting EGF-R.
Dai
Fei-Han
FH
National Laboratory for Oncogenes and Related Genes, Shanghai Cancer Institute, 25/Ln 2200, Xie Tu Road, 200032, Shanghai, P.R. China.
Chen
Yan
Y
Ren
Chang-Chun
CC
Li
Jin-Jun
JJ
Yao
Min
M
Han
Jun-Song
JS
Gong
Yi
Y
Yang
Sheng-Li
SL
Zhu
Jing-De
JD
Gu
Jian-Ren
JR
D000818:Animals; D001859:Bone Neoplasms / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000628:therapy*; D066246:ErbB Receptors / Q000378:metabolism*; D004926:Escherichia coli; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006657:Histones / Q000235:genetics*; D006801:Humans; D007150:Immunohistochemistry; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D015514:Oncogene Proteins, Fusion / Q000032:analysis / Q000302:isolation & purification; D012516:Osteosarcoma / Q000201:enzymology / Q000235:genetics / Q000378:metabolism* / Q000628:therapy*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured; D001616:beta-Galactosidase / Q000378:metabolism
D006657:Histones; D015514:Oncogene Proteins, Fusion; D066246:ErbB Receptors; D001616:beta-Galactosidase