Efficient drug screening and gene correction for treating liver disease using patient-specific stem cells.
Efficient drug screening and gene correction for treating liver disease using patient-specific stem cells.
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DOI：
10.1002/hep.26237
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发表时间：
2013-06
期刊：
影响因子：
13.5
通讯作者：
Jang, Yoon-Young
中科院分区：
文献类型：
--
作者：
Choi, Su Mi;Kim, Yonghak;Shim, Joong Sup;Park, Joon Tae;Wang, Rui-Hong;Leach, Steven D.;Liu, Jun O.;Deng, Chuxia;Ye, Zhaohui;Jang, Yoon-Young
患者特异性诱导多能干细胞(IPSCs)是开发新型药物和细胞疗法的潜在来源。尽管产生了越来越多的疾病特异性IPSCs，但基于IPSC的肝病药物筛选/发现进展有限，且人IPSCs的基因靶向效率较低，需要进一步改进。使用来自α-1抗胰蛋白酶(AAT)缺乏症患者的IPSC株，目前尚无药物或基因治疗可用，我们建立了一个平台，以发现新的候选药物并高效纠正致病突变。基于我们的肝脏分化方案实施了高通量格式筛选试验，以便于使用96孔免疫荧光读取器自动定量细胞AAT积聚。为了加快先导化合物对患者的最终应用，我们利用我们建立的临床化合物库--约翰·霍普金斯药物图书馆--进行了药物筛选，该库具有广泛的安全性概况。通过盲目大规模药物筛选，确定了五种临床药物，以减少不同患者IPSC来源的肝细胞样细胞中AAT的积聚。此外，利用最近开发的转录激活物样效应核酸酶(TALEN)技术，我们在AAT缺乏症患者IPSCs中实现了高基因靶向效率，25%-33%的克隆同时靶向两个患病等位基因。从基因校正的ipSCs获得的肝细胞样细胞具有功能，没有突变的AAT积聚。这种高效率和高性价比的目标定位技术将使基础和翻译应用程序广泛受益。结论：我们的结果证明了利用基于IPSC的疾病模型进行有效的大规模药物筛选的可行性，以及在人类IPSC中高度稳定的基因靶向；这两者对于将IPSC技术转化为治疗无法治疗的疾病的新疗法至关重要。
Patient-specific induced pluripotent stem cells (iPSCs) represent a potential source for developing novel drugand cell- therapies. Although increasing numbers of disease-specific iPSCs have been generated, there has been limited progress in iPSC-based drug screening/discovery for liver diseases, and the low gene targeting efficiency in human iPSCs warrants further improvement. Using iPSC lines from patients with alpha-1 antitrypsin (AAT) deficiency, for which there is currently no drug- or gene- therapy available, we established a platform to discover new drug candidates and to correct disease-causing mutation with a high efficiency. A high-throughput format screening assay based on our hepatic differentiation protocol was implemented to facilitate automated quantification of cellular AAT accumulation using a 96-well immunofluorescence reader. To expedite the eventual application of lead compounds to patients, we conducted drug screening utilizing our established library of clinical compounds, the Johns Hopkins Drug Library, with extensive safety profiles. Through a blind large-scale drug screening, five clinical drugs were identified to reduce AAT accumulation in diverse patient iPSC-derived hepatocyte-like cells. In addition, using the recently developed transcription activator-like effector nuclease (TALEN) technology, we achieved high gene targeting efficiency in AAT-deficiency patient iPSCs with 25–33% of the clones demonstrating simultaneous targeting at both diseased alleles. The hepatocyte-like cells derived from the gene-corrected iPSCs were functional without the mutant AAT accumulation. This highly efficient and cost-effective targeting technology will broadly benefit both basic and translational applications. Conclusions: Our results demonstrated the feasibility of effective large-scale drug screening using an iPSC-based disease model and highly robust gene targeting in human iPSCs; both of which are critical for translating the iPSC technology into novel therapies for untreatable diseases.
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影响因子：
46.9
作者：
Hockemeyer, Dirk;Soldner, Frank;Beard, Caroline;Gao, Qing;Mitalipova, Maisam;DeKelver, Russell C.;Katibah, George E.;Amora, Ranier;Boydston, Elizabeth A.;Zeitler, Bryan;Meng, Xiangdong;Miller, Jeffrey C.;Zhang, Lei;Rebar, Edward J.;Gregory, Philip D.;Urnov, Fyodor D.;Jaenisch, Rudolf
通讯作者：
Jaenisch, Rudolf
影响因子：
14.9
作者：
Cade L;Reyon D;Hwang WY;Tsai SQ;Patel S;Khayter C;Joung JK;Sander JD;Peterson RT;Yeh JR
通讯作者：
Yeh JR
影响因子：
17.1
作者：
Liu H;Kim Y;Sharkis S;Marchionni L;Jang YY
通讯作者：
Jang YY
影响因子：
4.8
作者：
Bouchecareilh, Marion;Hutt, Darren M.;Balch, William E.
通讯作者：
Balch, William E.
使用表达易于聚集的丝氨酸蛋白酶抑制剂 α1-抗胰蛋白酶 Z 的秀丽隐杆线虫进行自动化高内涵活体动物药物筛选。
影响因子：
3.7
作者：
Gosai SJ;Kwak JH;Luke CJ;Long OS;King DE;Kovatch KJ;Johnston PA;Shun TY;Lazo JS;Perlmutter DH;Silverman GA;Pak SC
通讯作者：
Pak SC