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PMID: 12475933 Published · ppublish English Journal Article

Protection against ischemic brain injury by protein therapeutics.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 99 ·No. 26 ·2002-12-24 ·Pages 17107-12

Asoh S, Ohsawa I, Mori T, Katsura K, Hiraide T, Katayama Y, Kimura M, Ozaki D, Yamagata K, Ohta S

Abstract

Preventing massive cell death is an important therapeutic strategy for various injuries and disorders. Protein therapeutics have the advantage of delivering proteins in a short period. We have engineered the antiapoptotic bcl-x gene to generate the super antiapoptotic factor, FNK, with a more powerful cytoprotective activity. In this study, we fused the protein transduction domain (PTD) of the HIVTat protein to FNK and used the construct in an animal model of ischemic brain injury. When added into culture media of human neuroblastoma cells and rat neocortical neurons, PTD-FNK rapidly transduced into cells and localized to mitochondria within 1 h. It protected the neuroblastomas and neurons against staurosporine-induced apoptosis and glutamate-induced excitotoxicity, respectively. The cytoprotective activity of PTD-FNK was found at concentrations as low as 0.3 pM. Additionally, PTD-FNK affected the cytosolic movement of calcium ions, which may relate to its neuroprotective action. Immunohistochemical analysis revealed that myc-tagged PTD-FNK (PTD-myc-FNK) injected i.p. into mice can have access into brain neurons. When injected i.p. into gerbils, PTD-FNK prevented delayed neuronal death in the hippocampus caused by transient global ischemia. These results suggest that PTD-FNK has a potential for clinical utility as a protein therapeutic strategy to prevent cell death in the brain.

MeSH Terms
Animals Apoptosis Brain Ischemia/drug therapy,pathology Cells, Cultured Gene Products, tat/chemistry Gerbillinae Humans Male Neuroblastoma/metabolism,pathology Neuroprotective Agents/therapeutic use Proto-Oncogene Proteins c-bcl-2/therapeutic use Recombinant Fusion Proteins/therapeutic use Tumor Cells, Cultured bcl-X Protein
Chemicals
BCL2L1 protein, human Gene Products, tat Neuroprotective Agents Proto-Oncogene Proteins c-bcl-2 Recombinant Fusion Proteins bcl-X Protein
Authors & Affiliations
10 authors, click to expand affiliations / ORCID
Asoh Sadamitsu
Department of Biochemistry and Cell Biology, Institute of Gerontology, Nippon Medical School, Kawasaki-city, Kanagawa 211-8533, Japan.
Ohsawa Ikuroh
Mori Takashi
Katsura Ken-Ichiro
Hiraide Tomoharu
Katayama Yasuo
Kimura Megumi
Ozaki Daiya
Yamagata Kumi
Ohta Shigeo
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.262460299
Published
2002-12-24
Epub
2002-00-10
Pages
17107-12
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC139277
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.262460299 PMC Full Text View journal details
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