BASIC RESEARCH FOR GENE THERAPY OF AMYOTROPHIC LATERAL SCLEROSIS USING ADENOVIRAL VECTORS
| Project/Area Number |
12670611
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurology
|
|---|
| Research Institution | KUMAMOTO UNIVERSITY |
|---|
Principal Investigator |
MITA Shuji KUMAMOTO UNIVERSITY HOSPITAL, NEUROLOGY, LECTURER, 医学部・附属病院, 講師 (30231616)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
UCHINO Makoto KUMAMOTO UNIVERSITY HOSPITAL, NEUROLOGY, PROFESSOR, 医学部・附属病院, 教授 (20117336)
|
|---|
| Project Period (FY) |
2000 – 2001
|
| Project Status |
Completed (Fiscal Year 2001)
|
| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2000: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | amyotrophic lateral sclerosis(ALS) / mutant SOD1 transgenic mouse / adenoviral vectors / retrograde transport / gene therapy / Bcl-2 / Cre-loxP recombination / protective effect on degeneration of motor neurons / Cre-loxp / 変異SOD1 transgenic mice |
|---|
| Research Abstract |
The possibility of somatic gene therapy as a viable therapeutic approach for motor neuron diseases has been investigated in several ways. We investigated whether adenoviral vectors containing the gene of interest, which has a protective effect on motor neurons, could rescue the affected motor neurons. In our initial study, we found that a marker gene of adenoviral vectors containing LacZ (AdLacZ)is expressed by retrograde transport in the hypoglossal cranial nerve nuclei after inoculation of AdLacZ into the tongue muscles of mutant SOD1 (G93A) transgenic mice (G93A mice). Subsequently, we employed a molecular switching cassette for Bcl-2 designed to express Bcl-2 by Cre-loxP recombination using adenoviral vectors, and examined in COS7 cells and primary neuronal cells with the mutant SOD1 gene. The overexpression of Bcl-2 in both cells and the neuronal protection against staurosporine-induced apoptosis were observed, after dual infection of adenoviral vectors with a cassette for Bcl-2 (
… More
AxCALNLBcl-2) and Cre recombinase (AxCANCre). After inoculation of AxCALNLBcl-2 followed by AxCANCre into the tongue of G93A mice,2 was detected in both the injection site and the hypoglossal nuclei of brainstems, suggesting the result of retrograde transport of AxCALNLBcl-2 and AxCANCre and expression of Bcl-2 by Cre recombinase in the hypoglossal nuclei. We examined the morphological changes of motor neurons in the hypoglossal nuclei of each mouse at 25 weeks of age, at which time the G93A mice manifested clinical signs, after inoculation of both vectors into the tongue of the G93A mice at 10 weeks of age. We found that the number of motor neurons was ignificantly higher in the G93A mice with both vectors than in those with AxCALNLBcl-2 alone or without inoculation. Further, we observed obvious reduction of vacuole formations and reactive astrocytes in and around the hypoglossal nuclei of G93A mice with both vectors in comparison with other groups of G93A mice. These results suggest that expression of Bcl-2 introduced by our system has not only a protective effect on degeneration of motor neurons but also an inhibitory effect on gliosis in G93A mice. This strategy for delivery of exogenous genes such as Bcl-2 will be useful in gene therapy for motor neuron diseases such as ALS. Less
|
Report
(3 results)
Research Products
(10 results)
