Protection Against Ischemic Brain Injury by Protein Therapeutics

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 2002 Dec 12

PMID 12475933

Citations 36

Authors

Sadamitsu Asoh

Ikuroh Ohsawa

Takashi Mori

Ken-ichiro Katsura

Tomoharu Hiraide

Yasuo Katayama

Megumi Kimura

Daiya Ozaki

Kumi Yamagata

Shigeo Ohta

Affiliations

Soon will be listed here.

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.

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References

Choi D . Glutamate neurotoxicity and diseases of the nervous system. Neuron. 1988; 1(8):623-34. DOI: 10.1016/0896-6273(88)90162-6. View

Busto R, Dietrich W, Globus M, Ginsberg M . Postischemic moderate hypothermia inhibits CA1 hippocampal ischemic neuronal injury. Neurosci Lett. 1989; 101(3):299-304. DOI: 10.1016/0304-3940(89)90549-1. View

Boise L, Gonzalez-Garcia M, Postema C, Ding L, LINDSTEN T, Turka L . bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death. Cell. 1993; 74(4):597-608. DOI: 10.1016/0092-8674(93)90508-n. View

Fawell S, Seery J, Daikh Y, Moore C, Chen L, Pepinsky B . Tat-mediated delivery of heterologous proteins into cells. Proc Natl Acad Sci U S A. 1994; 91(2):664-8. PMC: 43009. DOI: 10.1073/pnas.91.2.664. View

Nitatori T, Sato N, Waguri S, Karasawa Y, Araki H, Shibanai K . Delayed neuronal death in the CA1 pyramidal cell layer of the gerbil hippocampus following transient ischemia is apoptosis. J Neurosci. 1995; 15(2):1001-11. PMC: 6577848. View

Linnik M, Zahos P, Geschwind M, Federoff H . Expression of bcl-2 from a defective herpes simplex virus-1 vector limits neuronal death in focal cerebral ischemia. Stroke. 1995; 26(9):1670-4; discussion 1675. DOI: 10.1161/01.str.26.9.1670. View

Mizuguchi M, Sohma O, Takashima S, Ikeda K, Yamada M, Shiraiwa N . Immunochemical and immunohistochemical localization of Bcl-x protein in the rat central nervous system. Brain Res. 1996; 712(2):281-6. DOI: 10.1016/0006-8993(95)01453-5. View

Reed J . Double identity for proteins of the Bcl-2 family. Nature. 1997; 387(6635):773-6. DOI: 10.1038/42867. View

Tsujimoto Y, Shimizu S, Eguchi Y, Kamiike W, Matsuda H . Bcl-2 and Bcl-xL block apoptosis as well as necrosis: possible involvement of common mediators in apoptotic and necrotic signal transduction pathways. Leukemia. 1997; 11 Suppl 3:380-2. View

10.

Ohsawa I, Takamura C, Kohsaka S . The amino-terminal region of amyloid precursor protein is responsible for neurite outgrowth in rat neocortical explant culture. Biochem Biophys Res Commun. 1997; 236(1):59-65. DOI: 10.1006/bbrc.1997.6903. View

11.

Lawrence M, McLaughlin J, Sun G, Ho D, McIntosh L, Kunis D . Herpes simplex viral vectors expressing Bcl-2 are neuroprotective when delivered after a stroke. J Cereb Blood Flow Metab. 1997; 17(7):740-4. DOI: 10.1097/00004647-199707000-00003. View

12.

ARITOMI M, Kunishima N, Inohara N, Ishibashi Y, Ohta S, Morikawa K . Crystal structure of rat Bcl-xL. Implications for the function of the Bcl-2 protein family. J Biol Chem. 1997; 272(44):27886-92. DOI: 10.1074/jbc.272.44.27886. View

13.

Chen J, Nagayama T, Jin K, Stetler R, Zhu R, Graham S . Induction of caspase-3-like protease may mediate delayed neuronal death in the hippocampus after transient cerebral ischemia. J Neurosci. 1998; 18(13):4914-28. PMC: 6792571. View

14.

Montal M . Mitochondria, glutamate neurotoxicity and the death cascade. Biochim Biophys Acta. 1998; 1366(1-2):113-26. DOI: 10.1016/s0005-2728(98)00124-8. View

15.

Nagahara H, Snyder E, Ho A, Latham D, Lissy N, Ezhevsky S . Transduction of full-length TAT fusion proteins into mammalian cells: TAT-p27Kip1 induces cell migration. Nat Med. 1998; 4(12):1449-52. DOI: 10.1038/4042. View

16.

Yamada M, Oligino T, Mata M, Goss J, Glorioso J, Fink D . Herpes simplex virus vector-mediated expression of Bcl-2 prevents 6-hydroxydopamine-induced degeneration of neurons in the substantia nigra in vivo. Proc Natl Acad Sci U S A. 1999; 96(7):4078-83. PMC: 22423. DOI: 10.1073/pnas.96.7.4078. View

17.

Antonawich F, Federoff H, Davis J . BCL-2 transduction, using a herpes simplex virus amplicon, protects hippocampal neurons from transient global ischemia. Exp Neurol. 1999; 156(1):130-7. DOI: 10.1006/exnr.1998.7004. View

18.

Ohsawa I, Takamura C, Morimoto T, Ishiguro M, Kohsaka S . Amino-terminal region of secreted form of amyloid precursor protein stimulates proliferation of neural stem cells. Eur J Neurosci. 1999; 11(6):1907-13. DOI: 10.1046/j.1460-9568.1999.00601.x. View

19.

Schwarze S, Ho A, Dowdy S . In vivo protein transduction: delivery of a biologically active protein into the mouse. Science. 1999; 285(5433):1569-72. DOI: 10.1126/science.285.5433.1569. View

20.

Asoh S, Mori T, Ohta S . Practicable recovery of plasmids carrying cDNAs encoding anti-apoptotic factors that had been introduced by particle bombardment into 5-week old rat hippocampal slices. Neurosci Lett. 1999; 272(1):62-6. DOI: 10.1016/s0304-3940(99)00485-1. View