Tabar V, Studer L: Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson's disease

Center for Stem Cell Biology, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York 10065, USA.
Nature (Impact Factor: 41.46). 11/2011; 480(7378):547-51. DOI: 10.1038/nature10648
Source: PubMed


Human pluripotent stem cells (PSCs) are a promising source of cells for applications in regenerative medicine. Directed differentiation of PSCs into specialized cells such as spinal motoneurons or midbrain dopamine (DA) neurons has been achieved. However, the effective use of PSCs for cell therapy has lagged behind. Whereas mouse PSC-derived DA neurons have shown efficacy in models of Parkinson's disease, DA neurons from human PSCs generally show poor in vivo performance. There are also considerable safety concerns for PSCs related to their potential for teratoma formation or neural overgrowth. Here we present a novel floor-plate-based strategy for the derivation of human DA neurons that efficiently engraft in vivo, suggesting that past failures were due to incomplete specification rather than a specific vulnerability of the cells. Midbrain floor-plate precursors are derived from PSCs 11 days after exposure to small molecule activators of sonic hedgehog (SHH) and canonical WNT signalling. Engraftable midbrain DA neurons are obtained by day 25 and can be maintained in vitro for several months. Extensive molecular profiling, biochemical and electrophysiological data define developmental progression and confirm identity of PSC-derived midbrain DA neurons. In vivo survival and function is demonstrated in Parkinson's disease models using three host species. Long-term engraftment in 6-hydroxy-dopamine-lesioned mice and rats demonstrates robust survival of midbrain DA neurons derived from human embryonic stem (ES) cells, complete restoration of amphetamine-induced rotation behaviour and improvements in tests of forelimb use and akinesia. Finally, scalability is demonstrated by transplantation into parkinsonian monkeys. Excellent DA neuron survival, function and lack of neural overgrowth in the three animal models indicate promise for the development of cell-based therapies in Parkinson's disease.

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Available from: M. Flint Beal, Oct 09, 2015
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    • "The differentiations into FB, MB and HB were monitored using qRT-PCR, and we confirmed the enrichment of the FACS-isolated populations with a selection of markers previously used to characterize regionalization of differentiated hESCs (Fig. 2C-J) (Kirkeby et al., 2012b). To obtain enriched populations of MB FP progenitors (which do not express SOX1), we added the SHH agonist Purmorphamine (Pur) to the cultures from day 0 to 9 for efficient ventralization of the cells (Kriks et al., 2011). In the presence of Pur, the expression of GFP was heavily decreased in all three conditions (Fig. 2B; supplementary material Fig. S3A-F), and, within the MB and HB cultures, a significant proportion of the cells were identified as CORIN + and FOXA2 + (supplementary material Fig. S3J-Q), consistent with a floor-plate structure in these regions of the embryo (Shimamura et al., 1995). "
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    ABSTRACT: MicroRNAs (miRNAs) have been implicated in regulating multiple processes during brain development in various species. However, the function of miRNAs in human brain development remains largely unexplored. Here, we provide a comprehensive analysis of miRNA expression of regionalized neural progenitor cells derived from human embryonic stem cells and human foetal brain. We found miR-92b-3p and miR-130b-5p to be specifically associated with neural progenitors and several miRNAs that display both age-specific and region-specific expression patterns. Among these miRNAs, we identified miR-10 to be specifically expressed in the human hindbrain and spinal cord, while being absent from rostral regions. We found that miR-10 regulates a large number of genes enriched for functions including transcription, actin cytoskeleton and ephrin receptor signalling. When overexpressed, miR-10 influences caudalization of human neural progenitor cells. Together, these data confirm a role for miRNAs in establishing different human neural progenitor populations. This dataset also provides a comprehensive resource for future studies investigating the functional role of different miRNAs in human brain development.
    Development 09/2015; DOI:10.1242/dev.122747 · 6.46 Impact Factor
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    • "For instance, iPSCderived neurons have been shown to match the first trimester stage of human development (Mariani et al., 2012). In the case of midbrain dopamine neurons, a cell type that requires approximately 30 days for cell fate specification, full maturation requires periods of several months to generate functional neurons capable of rescuing dopamine deficits in animal models of PD upon transplantation (Kriks et al., 2011). The process of maturation is distinct from aging and is defined as the gain of full functionality of a given cell or tissue. "
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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.
    Development 09/2015; 142(18):3085-3089. DOI:10.1242/dev.120667 · 6.46 Impact Factor
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    • "Their study using PSCs has revealed interesting features of human neuronal development, in particular regarding their maturation kinetics, which suggest that neurons mature along their own, species-specific 'clock'. Indeed, human neurons display much more prolonged patterns of morphological and electrophysiological maturation than their mouse counterparts (Espuny-Camacho et al., 2013; Kriks et al., 2011; Maroof et al., 2013; Nicholas et al., 2013; Shi et al., 2012; Studer et al., 2015; Takazawa et al., 2012). For instance, whereas mouse cortical neurons typically mature in 2-4 weeks in vitro (Gaspard et al., 2008), in vitro derived human cortical neurons exhibit immature profiles of gene expression and excitability for several weeks. "
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    ABSTRACT: The human brain is arguably the most complex structure among living organisms. However, the specific mechanisms leading to this complexity remain incompletely understood, primarily because of the poor experimental accessibility of the human embryonic brain. Over recent years, technologies based on pluripotent stem cells (PSCs) have been developed to generate neural cells of various types. While the translational potential of PSC technologies for disease modeling and/or cell replacement therapies is usually put forward as a rationale for their utility, they are also opening novel windows for direct observation and experimentation of the basic mechanisms of human brain development. PSC-based studies have revealed that a number of cardinal features of neural ontogenesis are remarkably conserved in human models, which can be studied in a reductionist fashion. They have also revealed species-specific features, which constitute attractive lines of investigation to elucidate the mechanisms underlying the development of the human brain, and its link with evolution.
    Development 09/2015; 142(18):3138-3150. DOI:10.1242/dev.120568 · 6.46 Impact Factor
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