通过间充质干细胞调节线粒体 NDUFS4 缺乏症中的氧化磷酸化和氧化还原稳态。
Modulation of oxidative phosphorylation and redox homeostasis in mitochondrial NDUFS4 deficiency via mesenchymal stem cells.
作者信息
Melcher Marlen, Danhauser Katharina, Seibt Annette, Degistirici Özer, Baertling Fabian, Kondadi Arun Kumar, Reichert Andreas S, Koopman Werner J H, Willems Peter H G M, Rodenburg Richard J, Mayatepek Ertan, Meisel Roland, Distelmaier Felix
机构信息
Department of General Pediatrics, Neonatology and Pediatric Cardiology, University Children's Hospital, Heinrich Heine University Düsseldorf, Moorenstraße 5, 40225, Düsseldorf, Germany.
Division of Pediatric Stem Cell Therapy, Clinic for Pediatric Oncology, Hematology and Clinical Immunology, Medical Faculty, Heinrich Heine University Medical Center, Düsseldorf, Germany.
出版信息
Stem Cell Res Ther. 2017 Jun 24;8(1):150. doi: 10.1186/s13287-017-0601-7.
BACKGROUND
Disorders of the oxidative phosphorylation (OXPHOS) system represent a large group among the inborn errors of metabolism. The most frequently observed biochemical defect is isolated deficiency of mitochondrial complex I (CI). No effective treatment strategies for CI deficiency are so far available. The purpose of this study was to investigate whether and how mesenchymal stem cells (MSCs) are able to modulate metabolic function in fibroblast cell models of CI deficiency.
METHODS
We used human and murine fibroblasts with a defect in the nuclear DNA encoded NDUFS4 subunit of CI. Fibroblasts were co-cultured with MSCs under different stress conditions and intercellular mitochondrial transfer was assessed by flow cytometry and fluorescence microscopy. Reactive oxygen species (ROS) levels were measured using MitoSOX-Red. Protein levels of CI were analysed by blue native polyacrylamide gel electrophoresis (BN-PAGE).
RESULTS
Direct cellular interactions and mitochondrial transfer between MSCs and human as well as mouse fibroblast cell lines were demonstrated. Mitochondrial transfer was visible in 13.2% and 6% of fibroblasts (e.g. fibroblasts containing MSC mitochondria) for human and mouse cell lines, respectively. The transfer rate could be further stimulated via treatment of cells with TNF-α. MSCs effectively lowered cellular ROS production in NDUFS4-deficient fibroblast cell lines (either directly via co-culture or indirectly via incubation of cell lines with cell-free MSC supernatant). However, CI protein expression and activity were not rescued by MSC treatment.
CONCLUSION
This study demonstrates the interplay between MSCs and fibroblast cell models of isolated CI deficiency including transfer of mitochondria as well as modulation of cellular ROS levels. Further exploration of these cellular interactions might help to develop MSC-based treatment strategies for human CI deficiency.
背景
氧化磷酸化（OXPHOS）系统紊乱是先天性代谢缺陷中的一大类。最常见的生化缺陷是线粒体复合物I（CI）单独缺乏。目前尚无针对CI缺乏的有效治疗策略。本研究的目的是调查间充质干细胞（MSC）是否以及如何调节CI缺乏的成纤维细胞模型中的代谢功能。
方法
我们使用了核DNA编码的CI亚基NDUFS4有缺陷的人和小鼠成纤维细胞。在不同应激条件下将成纤维细胞与MSC共培养，并通过流式细胞术和荧光显微镜评估细胞间线粒体转移。使用MitoSOX-Red测量活性氧（ROS）水平。通过蓝色原代聚丙烯酰胺凝胶电泳（BN-PAGE）分析CI的蛋白水平。
结果
证明了MSC与人以及小鼠成纤维细胞系之间存在直接的细胞相互作用和线粒体转移。对于人和小鼠细胞系，分别在13.2%和6%的成纤维细胞（例如含有MSC线粒体的成纤维细胞）中可见线粒体转移。通过用TNF-α处理细胞可进一步刺激转移率。MSC有效降低了NDUFS4缺陷成纤维细胞系中的细胞ROS产生（直接通过共培养或间接通过将细胞系与无细胞MSC上清液孵育）。然而，MSC处理未能挽救CI蛋白表达和活性。
结论
本研究证明了MSC与孤立CI缺乏的成纤维细胞模型之间的相互作用，包括线粒体转移以及细胞ROS水平的调节。对这些细胞相互作用的进一步探索可能有助于开发基于MSC的人类CI缺乏治疗策略。
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