Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson’s disease. Nature

Center for Stem Cell Biology, Memorial Sloan-Kettering Cancer Center, 1275 York Avenue, New York, New York 10065, USA.
Nature (Impact Factor: 42.35). 11/2011; 480(7378):547-51. DOI: 10.1038/nature10648
Source: PubMed

ABSTRACT 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, Aug 17, 2015
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    • "Recent studies of cell replacement therapy in the anterior CNS using hPSC-derived progenitors have demonstrated that the implanted cells must possess a regional phenotype that mimics endogenous tissues in order to effectively engraft and correct neural deficits (Kriks et al., 2011; Ma et al., 2012). Since HOX expression patterns are crucial determinants of cellular phenotype, organization, and neural circuit integration in the developing hindbrain and spinal cord (Philippidou and Dasen, 2013), our patterning approach could serve as the basis for generating a spectrum of posterior neural progeny with highly specific regional identities that may aid regenerative therapy efforts. "
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    ABSTRACT: Colinear HOX expression during hindbrain and spinal cord development diversifies and assigns regional neural phenotypes to discrete rhombomeric and vertebral domains. Despite the precision of HOX patterning in vivo, in vitro approaches for differentiating human pluripotent stem cells (hPSCs) to posterior neural fates coarsely pattern HOX expression thereby generating cultures broadly specified to hindbrain or spinal cord regions. Here, we demonstrate that successive activation of fibroblast growth factor, Wnt/β-catenin, and growth differentiation factor signaling during hPSC differentiation generates stable, homogenous SOX2(+)/Brachyury(+) neuromesoderm that exhibits progressive, full colinear HOX activation over 7 days. Switching to retinoic acid treatment at any point during this process halts colinear HOX activation and transitions the neuromesoderm into SOX2(+)/PAX6(+) neuroectoderm with predictable, discrete HOX gene/protein profiles that can be further differentiated into region-specific cells, e.g., motor neurons. This fully defined approach significantly expands capabilities to derive regional neural phenotypes from diverse hindbrain and spinal cord domains. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
    Stem Cell Reports 04/2015; 33(4). DOI:10.1016/j.stemcr.2015.02.018 · 5.64 Impact Factor
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    • "Brain developmental and functional processes are highly dependent on interactions between different neural cell types and between different regions of the brain (for example, De Marco Garcia et al., 2011; Nishi, 2003). A great deal of attention has, for example, been focused on differentiation of human mDA neurons from hPSCs (Kriks et al., 2011; Perrier et al., 2004; Vazin et al., 2009; Yan et al., 2005; Zeng et al., 2004) because of their potential for use in transplantation therapy for Parkinson's disease. The development and formation of mDA neurons does not, however, occur in isolation , and DA systems and their targets structures are highly interdependent (Halliday et al., 2000; Hemmendinger et al., 1981; Hoffman et al., 1983; Parish et al., 2001; Prasad and Pasterkamp, 2009; Shalaby et al., 1984). "
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    ABSTRACT: We describe a technique for independently differentiating neocortical and mesencephalic dopaminergic (mDA) neurons from a single human pluripotent stem cell (hPSC) line, and subsequently allowing the two cell types to interact and form connections. Dopaminergic and neocortical progenitors were differentiated in separate vessels, then separately seeded into the inner and outer compartments of specialized cell culture vessels designed for in vitro studies of wound healing. Cells were further differentiated using dopamine-specific and neocortex-specific trophic factors, respectively. The barrier was then removed, and differentiation was continued for three weeks in the presence of BDNF. After three weeks of differentiation, neocortical and mDA cell bodies largely remained in the areas into which they had been seeded, and the gap between the mDA and neocortical neuron populations could still be discerned. Abundant tyrosine hydroxylase (TH)-positive projections had extended from the area of the inner chamber to outer chamber neocortical area. We have developed a hPSC-based system for producing connections between neurons from two brain regions, neocortex and midbrain. Future experiments could employ modifications of this method to examine connections between any two brain regions or neuronal subtypes that can be produced from hPSCs in vitro.
    04/2015; 33(3). DOI:10.3233/RNN-140488
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    • "As such, TRANSEURO, a randomized controlled trial of fetal cell therapies was recently launched incorporating the lessons of prior experience (Abbott, 2014). In addition, based on excellent progress in preclinical studies in rodents and primates using human ESC-and iPSC-derived cells (Ganat et al., 2012; Kriks et al., 2011; Sundberg et al., 2013), it is anticipated that trials of human stem-cell-derived dopaminergic neurons will be initiated in the near future. "
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    ABSTRACT: Decisions about what experimental therapies are advanced to clinical trials are based almost exclusively on findings in preclinical animal studies. Over the past 30 years, animal models have forecast the success of hundreds of neuroprotective pharmacological therapies for stroke, Alzheimer׳s disease, spinal cord injury, traumatic brain injury and amyotrophic lateral sclerosis. Yet almost without exception, all have failed. Rapid advances in stem cell technologies have raised new hopes that these neurological diseases may one day be treatable. Still, how can neuroregenerative therapies be translated into clinical realities if available animal models are such poor surrogates of human disease? To address this question we discuss human and rodent neurogenesis, evaluate mechanisms of action for cellular therapies and describe progress in translating neuroregeneration to date. We conclude that not only are appropriate animal models critical to the development of safe and effective therapies, but that the multiple mechanisms of stem cell-mediated therapies may be particularly well suited to the mechanistically diverse nature of central nervous system diseases in mice and man. Copyright © 2015. Published by Elsevier B.V.
    European journal of pharmacology 03/2015; 759. DOI:10.1016/j.ejphar.2015.03.041 · 2.68 Impact Factor
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