In vivo delivery of glial cell-derived neurotrophic factor across the blood-brain barrier by gene transfer into brain capillary endothelial cells

Chen Jiang, Noriko Koyabu, Yoshikazu Yonemitsu, Takao Shimazoe, Shigenori Watanabe, Mikihiko Naito, Takashi Tsuruo, Hisakazu Ohtani, Yasufumi Sawada

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The gene encoding mouse glial cell-derived neurotrophic factor (mGDNF gene) was transfected into brain capillary endothelial cells (BCECs) with the aim of delivering the gene product extensively into the brain parenchyma by making use of the secretory function of BCECs. First, we transfected mGDNF gene into cultured BCECs (MBEC4; mouse brain capillary endothelial cells) in vitro. The amount of mGDNF protein secreted from the transfected cells into the medium was 1500 to 3200 pg/mg of cell protein per day, being about sevenfold higher than that accumulated intracellularly. Furthermore, the basolateral-directed secretion of mGDNF protein from the transfected MBEC4 cells was fivefold higher than the apical-directed secretion. Next, the hemagglutination virus of Japan (HVJ)-liposomes encapsulating mGDNF gene were administered to rats in vivo via the internal carotid artery. The transfected rats showed a marked increase in the brain level of GDNF as assessed by means of enzyme-linked immunosorbent assay (ELISA) and Western blotting on day 3 after the administration, and the level remained significantly elevated for at least 12 days. Furthermore, immunohistochemical staining revealed an increase in GDNF immunoreactivity throughout the transfected forebrain. These results indicate that the gene was successfully transferred in vivo from HVJ-liposomes into BCECs, where it was expressed, and the gene product was secreted into the brain. Then, using this delivery method, we evaluated the protective effect for dopamine neuron against a retrograde 6-hydroxydopamine (6-OHDA) lesion, as assessed by behavioral and neurochemical indices.

Original languageEnglish
Pages (from-to)1181-1191
Number of pages11
JournalHuman Gene Therapy
Volume14
Issue number12
DOIs
Publication statusPublished - 2003 Aug 10
Externally publishedYes

Fingerprint

Nerve Growth Factors
Blood-Brain Barrier
Neuroglia
Endothelial Cells
Brain
Genes
Glial Cell Line-Derived Neurotrophic Factor
Oxidopamine
Hemagglutination
Liposomes
Japan
Viruses
Proteins
Dopaminergic Neurons
Internal Carotid Artery
Prosencephalon
Western Blotting
Enzyme-Linked Immunosorbent Assay
Staining and Labeling

ASJC Scopus subject areas

  • Genetics

Cite this

In vivo delivery of glial cell-derived neurotrophic factor across the blood-brain barrier by gene transfer into brain capillary endothelial cells. / Jiang, Chen; Koyabu, Noriko; Yonemitsu, Yoshikazu; Shimazoe, Takao; Watanabe, Shigenori; Naito, Mikihiko; Tsuruo, Takashi; Ohtani, Hisakazu; Sawada, Yasufumi.

In: Human Gene Therapy, Vol. 14, No. 12, 10.08.2003, p. 1181-1191.

Research output: Contribution to journalArticle

Jiang, Chen ; Koyabu, Noriko ; Yonemitsu, Yoshikazu ; Shimazoe, Takao ; Watanabe, Shigenori ; Naito, Mikihiko ; Tsuruo, Takashi ; Ohtani, Hisakazu ; Sawada, Yasufumi. / In vivo delivery of glial cell-derived neurotrophic factor across the blood-brain barrier by gene transfer into brain capillary endothelial cells. In: Human Gene Therapy. 2003 ; Vol. 14, No. 12. pp. 1181-1191.
@article{1cf668308d00408a89a75c7c2485ec46,
title = "In vivo delivery of glial cell-derived neurotrophic factor across the blood-brain barrier by gene transfer into brain capillary endothelial cells",
abstract = "The gene encoding mouse glial cell-derived neurotrophic factor (mGDNF gene) was transfected into brain capillary endothelial cells (BCECs) with the aim of delivering the gene product extensively into the brain parenchyma by making use of the secretory function of BCECs. First, we transfected mGDNF gene into cultured BCECs (MBEC4; mouse brain capillary endothelial cells) in vitro. The amount of mGDNF protein secreted from the transfected cells into the medium was 1500 to 3200 pg/mg of cell protein per day, being about sevenfold higher than that accumulated intracellularly. Furthermore, the basolateral-directed secretion of mGDNF protein from the transfected MBEC4 cells was fivefold higher than the apical-directed secretion. Next, the hemagglutination virus of Japan (HVJ)-liposomes encapsulating mGDNF gene were administered to rats in vivo via the internal carotid artery. The transfected rats showed a marked increase in the brain level of GDNF as assessed by means of enzyme-linked immunosorbent assay (ELISA) and Western blotting on day 3 after the administration, and the level remained significantly elevated for at least 12 days. Furthermore, immunohistochemical staining revealed an increase in GDNF immunoreactivity throughout the transfected forebrain. These results indicate that the gene was successfully transferred in vivo from HVJ-liposomes into BCECs, where it was expressed, and the gene product was secreted into the brain. Then, using this delivery method, we evaluated the protective effect for dopamine neuron against a retrograde 6-hydroxydopamine (6-OHDA) lesion, as assessed by behavioral and neurochemical indices.",
author = "Chen Jiang and Noriko Koyabu and Yoshikazu Yonemitsu and Takao Shimazoe and Shigenori Watanabe and Mikihiko Naito and Takashi Tsuruo and Hisakazu Ohtani and Yasufumi Sawada",
year = "2003",
month = "8",
day = "10",
doi = "10.1089/104303403322168019",
language = "English",
volume = "14",
pages = "1181--1191",
journal = "Human Gene Therapy",
issn = "1043-0342",
publisher = "Mary Ann Liebert Inc.",
number = "12",

}

TY - JOUR

T1 - In vivo delivery of glial cell-derived neurotrophic factor across the blood-brain barrier by gene transfer into brain capillary endothelial cells

AU - Jiang, Chen

AU - Koyabu, Noriko

AU - Yonemitsu, Yoshikazu

AU - Shimazoe, Takao

AU - Watanabe, Shigenori

AU - Naito, Mikihiko

AU - Tsuruo, Takashi

AU - Ohtani, Hisakazu

AU - Sawada, Yasufumi

PY - 2003/8/10

Y1 - 2003/8/10

N2 - The gene encoding mouse glial cell-derived neurotrophic factor (mGDNF gene) was transfected into brain capillary endothelial cells (BCECs) with the aim of delivering the gene product extensively into the brain parenchyma by making use of the secretory function of BCECs. First, we transfected mGDNF gene into cultured BCECs (MBEC4; mouse brain capillary endothelial cells) in vitro. The amount of mGDNF protein secreted from the transfected cells into the medium was 1500 to 3200 pg/mg of cell protein per day, being about sevenfold higher than that accumulated intracellularly. Furthermore, the basolateral-directed secretion of mGDNF protein from the transfected MBEC4 cells was fivefold higher than the apical-directed secretion. Next, the hemagglutination virus of Japan (HVJ)-liposomes encapsulating mGDNF gene were administered to rats in vivo via the internal carotid artery. The transfected rats showed a marked increase in the brain level of GDNF as assessed by means of enzyme-linked immunosorbent assay (ELISA) and Western blotting on day 3 after the administration, and the level remained significantly elevated for at least 12 days. Furthermore, immunohistochemical staining revealed an increase in GDNF immunoreactivity throughout the transfected forebrain. These results indicate that the gene was successfully transferred in vivo from HVJ-liposomes into BCECs, where it was expressed, and the gene product was secreted into the brain. Then, using this delivery method, we evaluated the protective effect for dopamine neuron against a retrograde 6-hydroxydopamine (6-OHDA) lesion, as assessed by behavioral and neurochemical indices.

AB - The gene encoding mouse glial cell-derived neurotrophic factor (mGDNF gene) was transfected into brain capillary endothelial cells (BCECs) with the aim of delivering the gene product extensively into the brain parenchyma by making use of the secretory function of BCECs. First, we transfected mGDNF gene into cultured BCECs (MBEC4; mouse brain capillary endothelial cells) in vitro. The amount of mGDNF protein secreted from the transfected cells into the medium was 1500 to 3200 pg/mg of cell protein per day, being about sevenfold higher than that accumulated intracellularly. Furthermore, the basolateral-directed secretion of mGDNF protein from the transfected MBEC4 cells was fivefold higher than the apical-directed secretion. Next, the hemagglutination virus of Japan (HVJ)-liposomes encapsulating mGDNF gene were administered to rats in vivo via the internal carotid artery. The transfected rats showed a marked increase in the brain level of GDNF as assessed by means of enzyme-linked immunosorbent assay (ELISA) and Western blotting on day 3 after the administration, and the level remained significantly elevated for at least 12 days. Furthermore, immunohistochemical staining revealed an increase in GDNF immunoreactivity throughout the transfected forebrain. These results indicate that the gene was successfully transferred in vivo from HVJ-liposomes into BCECs, where it was expressed, and the gene product was secreted into the brain. Then, using this delivery method, we evaluated the protective effect for dopamine neuron against a retrograde 6-hydroxydopamine (6-OHDA) lesion, as assessed by behavioral and neurochemical indices.

UR - http://www.scopus.com/inward/record.url?scp=0042768492&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0042768492&partnerID=8YFLogxK

U2 - 10.1089/104303403322168019

DO - 10.1089/104303403322168019

M3 - Article

C2 - 12908969

AN - SCOPUS:0042768492

VL - 14

SP - 1181

EP - 1191

JO - Human Gene Therapy

JF - Human Gene Therapy

SN - 1043-0342

IS - 12

ER -