Neural stem cell systems: physiological players or in vitro entities?

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Induced pluripotent stem cell lines from Huntington's disease mice undergo neuronal differentiation while showing alterations in the lysosomal pathway

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Huntingtin gene evolution in Chordata and its peculiar features in the ascidian Ciona genus

BACKGROUND: To gain insight into the evolutionary features of the huntingtin (htt) gene in Chordata, we have sequenced and characterized the full-length htt mRNA in the ascidian Ciona intestinalis, a basal chordate emerging as new invertebrate model organism. Moreover, taking advantage of the availability of genomic and EST sequences, the htt gene structure of a number of chordate species, including the cogeneric ascidian Ciona savignyi, and the vertebrates Xenopus and Gallus was reconstructed. RESULTS: The C. intestinalis htt transcript exhibits some peculiar features, such as spliced leader trans-splicing in the 98 nt-long 5' untranslated region (UTR), an alternative splicing in the coding region, eight alternative polyadenylation sites, and no similarities of both 5' and 3'UTRs compared to homologs of the cogeneric C. savignyi. The predicted protein is 2946 amino acids long, shorter than its vertebrate homologs, and lacks the polyQ and the polyP stretches found in the the N-terminal regions of mammalian homologs. The exon-intron organization of the htt gene is almost identical among vertebrates, and significantly conserved between Ciona and vertebrates, allowing us to hypothesize an ancestral chordate gene consisting of at least 40 coding exons. CONCLUSION: During chordate diversification, events of gain/loss, sliding, phase changes, and expansion of introns occurred in both vertebrate and ascidian lineages predominantly in the 5'-half of the htt gene, where there is also evidence of lineage-specific evolutionary dynamics in vertebrates. On the contrary, the 3'-half of the gene is highly conserved in all chordates at the level of both gene structure and protein sequence. Between the two Ciona species, a fast evolutionary rate and/or an early divergence time is suggested by the absence of significant similarity between UTRs, protein divergence comparable to that observed between mammals and fishes, and different distribution of repetitive elements

Characterization, developmental expression and evolutionary features of the huntingtin gene in the amphioxus Branchiostoma floridae

<p>Abstract</p> <p>Background</p> <p>Huntington's disease is an inherited neurodegenerative disorder that is caused by the expansion of an N-terminal polyQ stretch in the huntingtin protein. In order to investigate the hypothesis that huntingtin was already involved in development of the nervous system in the last common ancestor of chordates, we isolated and characterised the huntingtin homologue from the amphioxus <it>Branchiostoma floridae</it>. In the present paper the amphioxus general term must be referred to <it>Branchiostoma floridae</it>.</p> <p>Results</p> <p>In this report, we show that the exon-intron organization of the amphioxus huntingtin gene is highly conserved with that of other vertebrates species. The AmphiHtt protein has two glutamine residues in the position of the typical vertebrate polyQ tract. Sequence conservation is greater along the entire length of the protein than in a previously identified <it>Ciona </it>huntingtin. The first three N-terminal HEAT repeats are highly conserved in vertebrates and amphioxus, although exon rearrangement has occurred in this region. <it>AmphiHtt </it>expression is detectable by in situ hybridization starting from the early neurula stage, where it is found in cells of the neural plate. At later stages, it is retained in the neural compartment but also it appears in limited and well-defined groups of non-neural cells. At subsequent larval stages, <it>AmphiHtt </it>expression is detected in the neural tube, with the strongest signal being present in the most anterior part.</p> <p>Conclusion</p> <p>The cloning of amphioxus huntingtin allows to infer that the polyQ in huntingtin was already present 540 million years ago and provides a further element for the study of huntingtin function and its evolution along the deuterostome branch.</p

Rescue of gene expression by modified REST decoy oligonucleotides in a cellular model of Huntington's disease

Transcriptional dysfunction is a prominent hallmark of Huntington's disease (HD). Several transcription factors have been implicated in the aetiology of HD progression and one of the most prominent is repressor element 1 (RE1) silencing transcription factor (REST). REST is a global repressor of neuronal gene expression and in the presence of mutant Huntingtin increased nuclear REST levels lead to elevated RE1 occupancy and a concomitant increase in target gene repression, including brain-derived neurotrophic factor. It is of great interest to devise strategies to reverse transcriptional dysregulation caused by increased nuclear REST and determine the consequences in HD. Thus far, such strategies have involved RNAi or mutant REST constructs. Decoys are double-stranded oligodeoxynucleotides corresponding to the DNA-binding element of a transcription factor and act to sequester it, thereby abrogating its transcriptional activity. Here, we report the use of a novel decoy strategy to rescue REST target gene expression in a cellular model of HD. We show that delivery of the decoy in cells expressing mutant Huntingtin leads to its specific interaction with REST, a reduction in REST occupancy of RE1s and rescue of target gene expression, including Bdnf. These data point to an alternative strategy for rebalancing the transcriptional dysregulation in HD

Systematic Chromosomal Analysis of Cultured Mouse Neural Stem Cell Lines

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Neuropotent self-renewing neural stem (NS) cells derived from mouse induced pluripotent stem (iPS) cells

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G-Force PD: a global initiative in coordinating stem cell-based dopamine treatments for Parkinson's disease.

Translating new cell-based therapies to the clinic for patients with neurodegenerative disorders is complex. It involves pre-clinical testing of the cellular product and discussions with several regulatory agencies, as well as ethical debates. In an attempt to support efforts around the world, we set up a global consortium that brings together the major funded teams working on developing a stem cell-derived neural transplantation therapy for Parkinson's disease (PD). This consortium, G-Force PD, involves teams from Europe, USA, and Japan, and has already met on two occasions to discuss common problems, solutions, and the roadmap to the clinic. In this short review, we lay out the brief history and rationale for this initiative and discuss some of the issues that arose in our most recent meeting (May 2015) as we consider undertaking first-in-human clinical trials with stem cell-derived neurons for PD.We would like to thank all the respective funding agencies that support our work in this area including the EU FP7 programmes that fund TRANSEURO (HEALTH-F5-2010-242003) and Neurostemcellrepair (HEALTH-2013-INNOVATION-1-602278), NYSTEM (C028503) that supports the work of Studer et al, and a grant from the Network Program for Realization of Regenerative Medicine from the Japan Agency for Medical Research and Development (AMED) that supports the work of Takahashi et al. We would also like to thank Parkinson’s UK for help funding the first GForce-PD meeting in London and the Parkinson’s Disease Foundation (PDF) and Weill Cornell for the second in New York.This is the final version of the article. It first appeared from NPG via http://dx.doi.org/10.1038/npjparkd.2015.1