TOP ＞ シンポジウム

シンポジウム Progress in the CNS non-clinical study achieved by iPS technique 2S5-1 The Next Phase of the NIH Center for Regenerative Medicine（NCRM）：Establishment of a Stem Cell Technology Facility Simeonov Anton National Center for Advancing Translational Sciences（NCATS）, NIH The field of stem cell technologies has not progressed from basic discovery to therapeutic application as efficiently as many had hoped. Few robust tools, technologies, protocols, and paradigms exist that allow researchers to reproducibly and efficiently maintain a stem cell population in a pluripotent state, produce pure populations of specific cell types, or influence endogenous stem cell populations in vivo. Multiple needs analyses by U.S. and international research bodies have concluded that the lack of such tools and technologies is what is currently limiting the progress of translational and clinical applications of stem cells, and that their creation and provision would be transformative to the stem cell field and to biomedicine. It is this need that the NIH/NCATS Stem Cell Technology Facility（SCTF）, a continuation of the NIH Center for Regenerative Medicine（NCRM）Common Fund Program, will address：the mission of the Facility is to tackle the top methodological or technical problems that currently impede therapeutic use of induced pluripotent stem cells and to rapidly deliver the resulting protocols, standards, data, and tool molecules to the public. The Facility will collaborate with researchers to validate the methods developed within for suitability in regenerative medicine applications. An overview of the facility will be presented, along with emerging small molecule screening technologies that NCATS has developed which can be leveraged to further advance the field of stem cell-derived therapies. 2S5-2 iPSC Non-Clinical Experiments for Nervous System（iNCENS）project-An attempt to evolve the CNS safety pharmacological evaluation by in vitro use of human-induced pluripotent stem cell-derived neurons Sato Kaoru Div Pharmacol, NIHS In the process of drug development, the improvement of predictability of the CNS safety pharmacological evaluation is required because the CNS adverse effects cause the drop-outs at the later stages of the developmental process. When human iPSC（hiPSC）-derived neurons were reported in 2007, in vitro use of hiPSC-derived neurons attracted attentions to overcome these issues. However, at present, the majority of the non-clinical CNS safety pharmacological tests are in vivo animal tests and the follow-up in vitro tests are performed when needed. The appearance of hiPSC-derived neurons made us realize that we had not discussed enough about the usefulness or validity of the in vitro evaluations in the CNS safety pharmacological evaluation. We therefore have just launched the“iNCENS”（iPSC non-clinical experiments for nervous system）project. This project has two aims, i.e., the establishment of reliable in vitro biomarkers for the risks of cognitive impairment and epilepsy, and the application of hiPSC-derived neurons to these in vitro systems. So far, we have found good candidates which can be used for the prediction of cognitive impairment and epilepsy. Furthermore, we have established the standard experimental protocol to select hiPSC-derived neurons suitable for the non-clinical CNS safety pharmacological evaluation and have found two lines of hiPSC-derived neurons that express NMDA receptors. We also have succeeded in recording synchronization of spontaneous unit activity of hiPSC-derived neurons across multi-recording channels. In this presentation, we will introduce the progress of our“across-institute”project and discuss about the potentials of the in vitro use of hiPSC-neurons in the CNS safety pharmacological evaluation. 2S5-3 Early stage development of human iPSCs-derived neurons and its application to non-clinical study Koganezawa Noriko，Ohara Yuki，Ootsu Mao，Shirao Tomoaki DNBB, Gunma University Graduate School of Medicine Recent advances in human induced pluripotent stem cells（hiPSCs）offer new possibilities for biomedical research and clinical applications. However, the detail processes of neuronal development from hiPSCs have not been known. In this study we analyzed development of hiPSCs-derived neurons（hiPS-neurons）, particularly focusing on their early developmental stages. We cultured iCell Neuron（Cellular Dynamics International）and compared their development with that of the primary cultured neurons derived from rat hippocampus. In 2 days in vitro（DIV）culture of the rat neurons, we observed three different stages which were stages 1, 2, and 3 in the developmental classification proposed by Dotti et al. （1988）. Most developed stage 3 neurons had several short neurites with one long neurite, which is destined for an axon. In the DIV2 iCell Neurons, we observed neurons in all stages similar to the rat neurons, although the number of stage 3 neurons was few. This result indicates that the hiPS-neurons differentiate slower than rat neurons. In addition, the length of the axons and the speed of axonal elongation were measured. We found iCell Neuron had significantly shorter axons and significantly slower elongation speed of axon. Finally we examined if this slow axonal elongation in iCell Neuron is because of the abnormality of the growth cones, and found no differences. Together, our study shows the growth of hiPS-neuronal axons is slower but its differentiation is comparably to rat neurons. Furthermore, in this symposium, I would like to mention about the use of hiPS-neurons as a tool for non-clinical study. Our preliminary data suggests that evaluation of hiPS-neuronal development could be a useful tool to assess developmental neurotoxicity of medicines. 2S5-4 The iPSC technology to model human neurodevelopmental diseases in vitro Sala Carlo，Montani Caterina，Vicidomini Cinzia，Verpelli Chiara CNR Neuroscience Institute and Department of Medical Biotechnology and Translational medicine, University of Milano Intellectual disability（ID）and autism spectrum disorders（ASD）are complex developmental mental disorders characterized by social and communicative deficits, language impairments and repetitive behaviors, with an estimated prevalence in Europe of 1-2%. Although several genetic alterations have been recognized as causal of ID and ASD, the etiopathogenesis of these complex diseases remain largely unknown.We investigated the molecular bases of ID and ASD by using an innovative strategy based on the recently established technology of genetic reprogramming. Because most of proteins encoded by genes involved in ID and ASD are associated to the synaptic junction between neurons and are involved in its function, we focused our attention on two genes that codify for the synaptic proteins Shank3 and IL1RAPL1. We generated iPS cells, which we induced to differentiation into excitatory and inhibitory neurons, from fibroblast of patient carrying mutations in these genes. Studying the function of these proteins in parallel with knock out mouse models for Shank3 and IL1RAPL1 we will contribute to better understand the molecular mechanisms of synapse formation, plasticity and learning and memory processes, and will open the possibility of future therapeutic approaches for neurodevelopmental diseases like ID and ASD.