Potential usefulness of D2R reporter gene imaging by IBF as gene therapy monitoring for cerebellar neurodegenerative diseases.
We investigated a gene expression imaging method to examine the level of therapeutic gene expression in the cerebellum. Using a human immunodeficiency virus derived lentivial vector, we expressed the dopamine D(2) receptor (D(2)R) as a reporter protein to mouse cerebellar Purkinje cells. Biodistribution and ex vivo autoradiography studies were performed by giving [(125)I]5-iodo-7-N-[(1-ethyl-2-pyrrolidinyl)methyl]carboxamide-2,3-dihydrobenzofuran ([(125)I]IBF) (1.85 MBq), as a radioactive D(2)R ligand, to model mice expressing the D(2)R with an HA tag (HA-D(2)R) in the cerebellum. In this study, [(125)I]IBF was bound to the D(2)R expressed in the cerebellum of the model mice selectively. Immunostaining was performed to confirm the HA-D(2)R expression in the cerebellum of the model mice. A significant correlation (r=0.900, P<0.001) between areas that expressed HA-D(2)R by immunostaining and areas in which [(125)I]IBF accumulated by the ex vivo autoradiograms was found. These results indicated that radioiodinated IBF is useful as a reporter probe to detect D(2)R reporter gene expression, which can be used for monitoring therapeutic gene expression in the cerebellum.
Shiba
Kazuhiro
K
Division of Tracer Kinetics, Advanced Science Research Center, Kanazawa University, Kanazawa, Ishikawa, Japan. shiba@med.kanazawa-u.ac.jp
Torashima
Takashi
T
Hirai
Hirokazu
H
Ogawa
Kazuma
K
Akhter
Nasima
N
Nakajima
Kenichi
K
Kinuya
Seigo
S
Mori
Hirofumi
H
D000818:Animals; D001572:Benzofurans; D002526:Cerebellar Diseases / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D016086:Lentivirus / Q000235:genetics; D051379:Mice; D008810:Mice, Inbred C57BL; D019636:Neurodegenerative Diseases / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D009930:Organic Chemicals; D011759:Pyrrolidines; D017448:Receptors, Dopamine D2 / Q000032:analysis* / Q000235:genetics / Q000378:metabolism
D001572:Benzofurans; D009930:Organic Chemicals; D011759:Pyrrolidines; D017448:Receptors, Dopamine D2; C062439:5-iodo-N-((1-ethyl-2-pyrrolidinyl)methyl)-2,3-dihydrobenzofurancarboxamide