2003 Fiscal Year Final Research Report Summary
Airway Inflammation due to the Mutated CFTR and Modulation with Macrolide Antibiotics
| Project/Area Number |
13670619
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Respiratory organ internal medicine
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| Research Institution | Okinaka Memorial Institute for Medical Research (2003)
Jikei University School of Medicine (2001-2002) |
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Principal Investigator |
YOSHIMURA Kunihiko Okinaka Memorial Institute for Medical Research, Researcher, 研究員 (60246452)
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| Co-Investigator(Kenkyū-buntansha) |
AKOI Kaoru Jikei University, School of Medicine, Assistant Professor, 医学部, 講師 (90212357)
GHANSHYAM D.Heda 米国Tennessee大学, 健康科学センター, 講師
HEDA Ghanshyam D. Tennessee University, Health Science Center, Assistant Professor
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| Project Period (FY) |
2001 – 2003
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| Keywords | CFTR / macrolides / gene mutations / diffuse panbronchiolitis / ΔF508 / cystic fibrosis / mutagenesis / gene expression |
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| Research Abstract |
Diffuse panbronchiolitis(DPB), a chronic inflammatory airway disease affecting mainly adults in Asian countries such as Japan, presents characteristic pulmonary manifestations similar to those of cystic fibrosis(CF). Since the introduction of macrolide antibiotics(MA) such as erythromycin and clarithromycin to treat patients with DPB has successfully and dramatically improved the prognosis of the disease, and our previous studies demonstrated a high prevalence of CFTR gene mutations in those patients with DPB, we have hypothesized that CFTR might be responsible, at least in part, for the pathogenesis of DPB, and the expression of CFTR could be modulated with MA as well. First, we have analyzed possible modulation of CFTR mRNA expression and splicing of CFTR exon 9 with MA. Interestingly, MA did not alter the levels of CFTR mRNA, but the amounts of exon 9-CFTR mRNA increased in a dose-dependent manner, suggesting the likely negative modulation of functional CFTR mRNA. In terms of CFTR channel property, LLCPK and LLCPKΔF508, which were stably transformed cell lines with normal CFTR cDNA and mutated CFTRΔF508, were utilized for in vitro chloride efflux assay. It seemed that MA could upregulate the function of CFTR channel activity. Next, we have continued to evaluate the CFTR genotypes in Japanese patients with CF, DPB and congenital bilateral absence of vas deferens. Through the study, we have found multiple rare or unique mutations which have not been deposited to the world-wide CF Mutation Database. Finally, the functional properties of the mutations detected in Japanese individuals such as 125C and Q1352H have been conducted by constructing expression plasmid vectors driven by Rous sarcoma virus promoter and stably transfecting those plasmids into CFTR-non-producing cells. The experiments and analyses are currently underway.
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