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Probing sporadic and familial Alzheimer's disease using induced pluripotent stem cells

机译:使用诱导性多能干细胞探查偶发性和家族性阿尔茨海默氏病

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摘要

Our understanding of Alzheimer's disease pathogenesis is currently limited by difficulties in obtaining live neurons from patients and the inability to model the sporadic form of the disease. It may be possible to overcome these challenges by reprogramming primary cells from patients into induced pluripotent stem cells (iPSCs). Here we repro-grammed primary fibroblasts from two patients with familial Alzheimer's disease, both caused by a duplication of the amyloid-β precursor protein gene (APP; termed APP~(Dp)), two with sporadic Alzheimer's disease (termed sAD1, sAD2) and two non-demented control individuals into iPSC lines. Neurons from differentiated cultures were purified with fluorescence-activated cell sorting and characterized. Purified cultures contained more than 90% neurons, clustered with fetal brain messenger RNA samples by microarray criteria, and could form functional synaptic contacts. Virtually all cells exhibited normal electrophysiological activity. Relative to controls, iPSC-derived, purified neurons from the two APP~(Dp) patients and patient sAD2 exhibited significantly higher levels of the pathological markers amyloid-P(l-40), phospho-tau(Thr 231) and active glycogen synthase kinase-3P (aGSK-3P). Neurons from APP~(Dp) and sAD2 patients also accumulated large RAB5-positive early endosomes compared to controls. Treatment of purified neurons with P-secretase inhibitors, but not γ-secretase inhibitors, caused significant reductions in phospho-Tau(Thr 231) and aGSK-3β levels. These results suggest a direct relationship between APP proteolytic processing, but not amyloid-P, in GSK-3β activation and tau phosphorylation in human neurons. Additionally, we observed that neurons with the genome of one sAD patient exhibited the phenotypes seen in familial Alzheimer's disease samples. More generally, we demonstrate that iPSC technology can be used to observe phenotypes relevant to Alzheimer's disease, even though it can take decades for overt disease to manifest in patients.
机译:目前,我们对阿尔茨海默氏病发病机理的理解受到从患者获得活神经元的困难以及无法对疾病的散发形式进行建模的限制。通过将患者的原代细胞重编程为诱导性多能干细胞(iPSC),有可能克服这些挑战。在这里,我们对两名患有家族性阿尔茨海默氏病的患者的原代成纤维细胞进行了重新编程,均由淀粉样蛋白-β前体蛋白基因(APP;称为APP〜(Dp))的重复引起;两名患有零星的阿尔茨海默氏病(称为sAD1,sAD2)和两个非痴呆的对照个体进入iPSC品系。来自分化培养物的神经元通过荧光激活细胞分选进行纯化和表征。纯化的培养物中含有超过90%的神经元,按照微阵列标准与胎儿脑信使RNA样本聚集在一起,并且可以形成功能性的突触接触。实际上,所有细胞均表现出正常的电生理活性。相对于对照,来自两名APP〜(Dp)患者和患者sAD2的iPSC纯化神经元显示出更高水平的病理标志物淀粉样蛋白P(l-40),磷酸化tau(Thr 231)和活性糖原合酶激酶3P(aGSK-3P)。与对照组相比,APP〜(Dp)和sAD2患者的神经元也积累了大量RAB5阳性的早期内体。用P分泌酶抑制剂而非γ分泌酶抑制剂处理纯化的神经元,可导致磷酸化Tau(Thr 231)和aGSK-3β水平显着降低。这些结果表明在人类神经元中GSK-3β激活中​​APP的蛋白水解过程而非淀粉样蛋白P与tau磷酸化之间存在直接关系。此外,我们观察到具有一名sAD患者基因组的神经元表现出在家族性阿尔茨海默氏病样本中看到的表型。更广泛地说,我们证明了iPSC技术可用于观察与阿尔茨海默氏病相关的表型,即使在患者身上出现明显的疾病可能需要数十年的时间。

著录项

  • 来源
    《Nature》 |2012年第7384期|p.216-220|共5页
  • 作者单位

    Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA,Biomedical Sciences Graduate Program, University of California, San Diego, La Jolla, California 92093, USA;

    Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA,Department of Neurosciences, University of California, San Diego, La Jolla, California, USA;

    The Salk Institute for Biological Studies, La Jolla, California 92037, USA;

    Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA,Biomedical Sciences Graduate Program, University of California, San Diego, La Jolla, California 92093, USA;

    The Salk Institute for Biological Studies, La Jolla, California 92037, USA;

    Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA;

    Department of Anesthesiology, University of California, San Diego, La Jolla, California 92093, USA;

    Department of Anesthesiology, Maastricht University Medical Center, Maastricht 6202 AZ, Netherlands;

    Departmentof Chemical Physiology, The Scripps Research Institute, La Jolla, California 92037, USA;

    Department of Reproductive Medicine, University of California, San Diego, La Jolla, California 92093, USA;

    BD Biosciences, La Jolla, California 92037, USA;

    Department of Reproductive Medicine, University of California, San Diego, La Jolla, California 92093, USA;

    Department of Anesthesiology, University of California, San Diego, La Jolla, California 92093, USA, Institute of Neurobiology, Slovak Academy of Sciences, Kosice SK-04001, Slovakia;

    The Salk Institute for Biological Studies, La Jolla, California 92037, USA;

    Department of Clinical Medicine, Neurology and Clinical Research Center, University of Oulu, Oulu FIN-90015, Finland;

    Department of Neurosciences, University of California, San Diego, La Jolla, California, USA;

    Howard Hughes Medical Institute and Department of Cellular and Molecular Medicine, University of California, San Diego, La Jolla, California 92093, USA,Department of Neurosciences, University of California, San Diego, La Jolla, California, USA;

  • 收录信息 美国《科学引文索引》(SCI);美国《工程索引》(EI);美国《生物学医学文摘》(MEDLINE);美国《化学文摘》(CA);
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