タイトル厚生労働科学研究費補助金（難治性疾患克服研究事業）「Menkes 病・occipital horn 症候群の実態調査、早期診断基準確立、治療法開発に関する研究」平成23-24年度 総合研究報告書

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厚生労働科学研究費補助金（難治性疾患克服研究事業）「Menkes 病・occipital horn 症候群の実態調査、早期診断基準確立、治療法開発に関する研究」平成23-24年度 総合研究報告書

Journal of Trace Elements in Medicine and Biology 26 (2012) 105?108Contents lists available at SciVerse ScienceDirectJournal of Trace Elements in Medicine and Biologyj ourna l homepage: www.elsevier.de/jtembEffect of copper and disulfiram combination therapy on the macular mouse, amodel of Menkes diseaseWattanaporn Bhadhprasit a , Hiroko Kodama a,b,? , Chie Fujisawa a , Tomoko Hiroki a , Eishin Ogawa aa Department of Pediatrics, Teikyo University School of Medicine, Tokyo, Japanb Department of Health and Dietetics, Faculty of Health and Medical Sciences, Teikyo Heisei University, Tokyo, Japana r t i c l e i n f oArticle history:Received 1 March 2012Accepted 30 April 2012Keywords:Menkes diseaseCopperDisulfiramCytochrome c oxidaseBlood?brain barriera b s t r a c tMenkes disease (MD) is a genetic neurodegenerative disorder characterized by copper deficiency due to adefect in ATP7A. Standard treatment involves parenteral copper?histidine administration. However, thetreatment is ineffective if initiated after two months of age, because the administered copper accumulatesin the blood?brain barrier and is not transported to neurons. To resolve this issue, we investigatedthe effects of a combination therapy comprising copper and disulfiram, a lipophilic chelator, in the macularmouse, an animal model of MD. Seven-day-old macular mice treated subcutaneously with 50 ?gof CuCl2 on postnatal day 4 were used. The mice were given a subcutaneous injection of CuCl2 (10 ?g)with oral administration of disulfiram (0.3 mg/g body weight) twice a week until eight weeks of age, andthen sacrificed. Copper concentrations in the cerebellum, liver, and serum of treated macular mice weresignificantly higher than those of control macular mice, which received only copper. Mice treated withthe combination therapy exhibited higher cytochrome c oxidase activity in the brain. The ratios of noradrenalineand adrenaline to dopamine in the brain were also increased by the treatment, suggesting thatdopamine ?-hydroxylase activity was improved by the combination therapy. Liver and renal functionswere almost normal, although renal copper concentration was higher in treated macular mice than incontrols. These results suggest that disulfiram facilitates the passage of copper across the blood?brainbarrier and that copper?disulfiram combination therapy may be an effective treatment for MD patients.c 2012 Elsevier GmbH. All rights reserved.IntroductionMenkes disease (MD) is an X-linked recessive disorder causedby a defect in a copper-transporting ATPase (ATP7A). In humans,ATP7A is expressed in almost all cell types except hepatocytes.In normal cells, ATP7A is localized in the trans-Golgi networkand transports copper from the cytosol into the Golgi apparatus,where copper is incorporated into secretory copper enzymes [1].In MD-affected cells, however, copper accumulates in the cytosoland cannot be excreted. Copper accumulation in the intestinesresults in copper absorption failure, leading to overall copper deficiencyin the body, except in the kidney. Copper also accumulatesin the cells of the blood?brain barrier. Thus, copper cannot bedelivered from the bloodstream to neurons after barrier maturation[2?4]. Copper concentrations in the serum, liver, and brainof MD patients are significantly lower, resulting in reduced activitiesof copper-dependent enzymes such as cytochrome c oxidase,dopamine ?-hydroxylase, and lysyl oxidase. Currently, parenteraladministration of copper?histidine is the standard treatment forMD [1,5]. However, the treatment is ineffective if initiated aftertwo months of age, because the administered copper accumulatesin the blood?brain barrier and is not transported to neurons. Therefore,copper delivery to neurons is the most important objective inthe treatment of MD-associated neurological degeneration [6].We previously reported that a combination therapy comprisingcopper and diethyldithiocarbonate (DEDTC) improved copperconcentrations, cytochrome c oxidase activity, and catecholaminemetabolism in the brain of macular mice [7]. A dimer of DEDTC,disulfiram, has been used for the treatment of alcoholism andcocaine addiction and as a modulator of cisplatin-induced toxicity[8?10]; thus, oral disulfiram is easily applicable in the clinicalsetting. Here, we report the effects of a combination therapy comprisingcopper injection and oral disulfiram on the macular mouse,an animal model of MD [11].Materials and methodsAnimals? Corresponding author at: Department of Health and Dietetics, Teikyo HeiseiUniversity, 2-51-4 Higashiikebukuro, Toshima-ku, Tokyo 170-8445, Japan.E-mail address: kodamah2018@gmail.com (H. Kodama).Male hemizygous macular mice and normal littermate controlswere treated with a single subcutaneous injection of cupric0946-672X/$ ? see front matter c 2012 Elsevier GmbH. All rights reserved.http://dx.doi.org/10.1016/j.jtemb.2012.05.00210