An AAV-derived Apaf-1 dominant negative inhibitor prevents MPTP toxicity as antiapoptotic gene therapy for Parkinson's disease

doi:10.1073/pnas.191107398

An AAV-derived Apaf-1 dominant negative inhibitor prevents MPTP toxicity as antiapoptotic gene therapy for Parkinson's disease

Departments of ^*Neurology and ^¶Hematology, Juntendo University School of Medicine, 2-1-1 Hongo, Bunkyo-ku, Tokyo 113-8421, Japan; ^‖Laboratory for Cell Recovery Mechanisms, The Institute of Physical and Chemical Research, Brain Science Institute, 2-1 Hirosawa, Wako, Saitama 351-0198, Japan; ^‡Department of Biochemistry and Molecular Biology, Nippon Medical School, 1-1-5 Sendagi, Bunkyo-ku, Tokyo 113-8602, Japan; and ^§Department of Neurology, Keio University School of Medicine, 35 Shinanomachi, Shinjyuku-ku, Tokyo 160-8582, Japan

Edited by H. Robert Horvitz, Massachusetts Institute of Technology, Cambridge, MA, and approved July 11, 2001 (received for review March 5, 2001)

Abstract

Adeno-associated virus (AAV) vector delivery of an Apaf-1-dominant negative inhibitor was tested for its antiapoptotic effect on degenerating nigrostriatal neurons in a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) model of Parkinson's disease. The wild-type caspase recruitment domain of Apaf-1 was used as a dominant negative inhibitor of Apaf-1 (rAAV-Apaf-1-DN-EGFP). An AAV virus vector was used to deliver it into the striatum of C57 black mice, and the animals were treated with MPTP. The number of tyrosine hydroxylase-positive neurons in the substantia nigra was not changed on the rAAV-Apaf-1-DN-EGFP injected side compared with the noninjected side. We also examined the effect of a caspase 1 C285G mutant as a dominant negative inhibitor of caspase 1 (rAAV-caspase-1-DN-EGFP) in the same model. However, there was no difference in the number of tyrosine hydroxylase-positive neurons between the rAAV-caspase-1-DN-EGFP injected side and the noninjected side. These results indicate that delivery of Apaf-1-DN by using an AAV vector system can prevent nigrostriatal degeneration in MPTP mice, suggesting that it could be a promising therapeutic strategy for patients with Parkinson's disease. The major mechanism of dopaminergic neuronal death triggered by MPTP seems to be the mitochondrial apoptotic pathway.

Footnotes

↵ † To whom reprint requests should be addressed. E-mail: hideki{at}med.juntendo.ac.jp.
This paper was submitted directly (Track II) to the PNAS office.
Abbreviations:

MPTP,

1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine;

TH,

tyrosine hydroxylase;

MTP,

mitochondrial transition pore;

AAV,

adeno-associated virus;

CARD,

caspase-recruitment domain;

EGFP,

enhanced green fluorescent protein;

PD,

Parkinson's disease