Mitochondrial iron and energetic dysfunction distinguish fibroblasts and induced neurons from pantothenate kinase-associated neurodegeneration patients

详细信息    查看全文

• 作者：Paolo Santambrogio^a ; Sabrina Dusi^b ; Michela Guaraldo^a ; ^c ; Luisa Ida Rotundo^a ; Vania Broccoli^a ; Barbara Garavaglia^b ; Valeria Tiranti^b ; Sonia Levi^a ; ^c ; ^{levi.sonia@hsr.it" class="auth_mail" title="E-mail the corresponding author}
• 关键词：DCF ; dichlorofluorescein ; DFO ; deferoxamine ; DHR-123 ; dihydrorhodamine 123 ; FAC ; ferric ammonium citrate ; iNs ; induced neurons ; IRP1 ; iron regulatory protein 1 ; LIP ; labile iron pool ; NBIA ; neurodegeneration with brain iron accumulation ; PKAN ; pantothenate kinase associated neurodegeneration ; PIH ; pyridoxal isonicotinoyl hydrazone ; ROS ; reactive oxygen species ; RPA ; rhodamine B-[(1 ; 10-phenanthroline-5-yl)-aminocarbonyl]benzyl ester ; RPAC ; rhodamine B-[(phenanthren-9-yl)-aminocarbonyl]-benzyleste
• 刊名：Neurobiology of Disease
• 出版年：2015
• 出版时间：September 2015
• 年：2015
• 卷：81
• 期：Complete
• 页码：144-153
• 全文大小：1303 K

文摘

Pantothenate kinase-associated neurodegeneration is an early onset autosomal recessive movement disorder caused by mutation of the pantothenate kinase-2 gene, which encodes a mitochondrial enzyme involved in coenzyme A synthesis. The disorder is characterised by high iron levels in the brain, although the pathological mechanism leading to this accumulation is unknown. To address this question, we tested primary skin fibroblasts from three patients and three healthy subjects, as well as neurons induced by direct fibroblast reprogramming, for oxidative status, mitochondrial functionality and iron parameters. The patients' fibroblasts showed altered oxidative status, reduced antioxidant defence, and impaired cytosolic and mitochondrial aconitase activities compared to control cells. Mitochondrial iron homeostasis and functionality analysis of patient fibroblasts indicated increased labile iron pool content and reactive oxygen species development, altered mitochondrial shape, decreased membrane potential and reduced ATP levels. Furthermore, analysis of induced neurons, performed at a single cell level, confirmed some of the results obtained in fibroblasts, indicating an altered oxidative status and signs of mitochondrial dysfunction, possibly due to iron mishandling. Thus, for the first time, altered biological processes have been identified in vitro in live diseased neurons. Moreover, the obtained induced neurons can be considered a suitable human neuronal model for the identification of candidate therapeutic compounds for this disease.