Generation of Isogenic Pluripotent Stem Cells Differing Exclusively at Two Early Onset Parkinson Point Mutations

The Whitehead Institute, 9 Cambridge Center, Cambridge, MA 02142, USA.
Cell (Impact Factor: 32.24). 07/2011; 146(2):318-31. DOI: 10.1016/j.cell.2011.06.019
Source: PubMed


Patient-specific induced pluripotent stem cells (iPSCs) derived from somatic cells provide a unique tool for the study of human disease, as well as a promising source for cell replacement therapies. One crucial limitation has been the inability to perform experiments under genetically defined conditions. This is particularly relevant for late age onset disorders in which in vitro phenotypes are predicted to be subtle and susceptible to significant effects of genetic background variations. By combining zinc finger nuclease (ZFN)-mediated genome editing and iPSC technology, we provide a generally applicable solution to this problem, generating sets of isogenic disease and control human pluripotent stem cells that differ exclusively at either of two susceptibility variants for Parkinson's disease by modifying the underlying point mutations in the α-synuclein gene. The robust capability to genetically correct disease-causing point mutations in patient-derived hiPSCs represents significant progress for basic biomedical research and an advance toward hiPSC-based cell replacement therapies.

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    • "mutations in genes such as α1-anti-trypsin (Yusa, et al., 2011), α-synuclein (SCNA;(Ryan, et al., 2013; Soldner, et al., 2011), and tau (MAPT(Fong, et al., 2013). However, the success of this technique is limited due to the challenging design of a robust engineered zinc finger nuclease (Hsu, et al., 2014; Ma, et al., 2015; Sander and Joung, 2014). "
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    ABSTRACT: In the last decade, induced pluripotent stem (iPS) cells have revolutionized the utility of human in vitro models of neurological disease. The iPS-derived and differentiated cells allow researchers to study the impact of a distinct cell type in health and disease as well as performing therapeutic drug screens on a human genetic background. In particular, clinical trials for Alzheimer's disease (AD) have been often failing. Two of the potential reasons are first, the species gap involved in proceeding from initial discoveries in rodent models to human studies, and second, an unsatisfying patient stratification, meaning subgrouping patients based on the disease severity due to the lack of phenotypic and genetic markers. iPS cells overcome this obstacles and will improve our understanding of disease subtypes in AD. They allow researchers conducting in depth characterization of neural cells from both familial and sporadic AD patients as well as preclinical screens on human cells. In this review, we briefly outline the status quo of iPS cell research in neurological diseases along with the general advantages and pitfalls of these models. We summarize how genome-editing techniques such as CRISPR/Cas will allow researchers to reduce the problem of genomic variability inherent to human studies, followed by recent iPS cell studies relevant to AD. We then focus on current techniques for the differentiation of iPS cells into neural cell types that are relevant to AD research. Finally, we discuss how the generation of three-dimensional cell culture systems will be important for understanding AD phenotypes in a complex cellular milieu, and how both two- and three-dimensional iPS cell models can provide platforms for drug discovery and translational studies into the treatment of AD.
    Full-text · Article · Dec 2015 · Molecular and Cellular Neuroscience
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    • "Indeed, such iPSC-based disease models are rapidly developing into a key platform for drug discovery and preclinical testing of candidate compounds (Lee et al., 2009, 2012a; Merkle and Eggan, 2013; Wainger et al., 2014). In the case of PD, recent studies have demonstrated that neurons derived from patient-specific iPSCs could successfully reproduce several disease-related phenotypes such as increased α-synuclein levels, mitochondrial dysfunction and hypersensitivity to toxins such as compounds that trigger mitochondrial stress or ROS (Byers et al., 2011; Devine et al., 2011; Seibler et al., 2011; Soldner et al., 2011; Cooper et al., 2012; Imaizumi et al., 2012; Liu et al., 2012; Sánchez-Danés et al., 2012; Miller et al., 2013; Reinhardt et al., 2013; Su and Qi, 2013; Sanders et al., 2014; Woodard et al., 2014). Most of these models showed evidence of biochemical changes that are directly dependent on the disease-specific genetic defects (Fig. 1). "
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    ABSTRACT: In contrast to the successful modeling of early-onset disorders using patient-specific cells, modeling of late-onset neurodegenerative diseases such as Parkinson's disease remains a challenge. This might be related to the often ignored fact that current induced pluripotent stem cell (iPSC) differentiation protocols yield cells that typically show the behavior of fetal stage cells. Acknowledging aging as a contributing factor in late-onset neurodegenerative disorders represents an important step on the road towards faithfully recreating these diseases in vitro. Here, we summarize progress in the field and review the strategies and challenges for triggering late-onset disease phenotypes.
    Full-text · Article · Sep 2015 · Development
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    • "Thus, if you find something different between the control and the patient-derived iPSCs you have to worry whether the difference is due to the pathology of the disease or to the variability between cell lines. For this reason, years ago, we started generating isogenic cell lines that differed from one another exclusively by a disease-causing mutation in order to generate a molecularly defined system in which we could study genetic diseases (Soldner et al., 2011). However, an important part of medically relevant diseases is that most are sporadic. "
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    ABSTRACT: Rudolf Jaenisch is a Professor of Biology at Massachusetts Institute of Technology, a founding member of the Whitehead Institute for Biomedical Research and the current president of the International Society for Stem Cell Research (ISSCR). His contributions to the stem cell field span from making the first transgenic mouse to seminal advances in the reprogramming field, and much more. In recognition of his pioneering research leading to induced pluripotency, he recently received the 2015 March of Dimes Prize in Developmental Biology. At the recent Keystone Meeting on 'Transcriptional and Epigenetic Influences on Stem Cell States' in Colorado, we had the opportunity to talk to him about his life and work. © 2015. Published by The Company of Biologists Ltd.
    Preview · Article · Jun 2015 · Development
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