10 research outputs found
Dgcr8 controls neural crest cells survival in cardiovascular development
No full textAbstractDiGeorge syndrome (DGS), characterized genetically by a deletion within chromosome 22q11.2, is associated with a constellation of congenital heart defects. DiGeorge critical region 8 (Dgcr8), a gene that maps to the common deletion region of DGS, encodes a double stranded RNA-binding protein that is essential for miRNA biogenesis. To address the potential contribution of Dgcr8 insufficiency to cardiovascular development, we have inactivated Dgcr8 in cardiac neural crest cells (cNCCs). Dgcr8 mutants displayed a wide spectrum of malformations, including persistent truncus arteriosus (PTA) and ventricular septal defect (VSD). Interestingly, Dgcr8-null cNCCs that properly migrated into the cardiac outflow tract (OFT), proliferate normally and differentiate into vascular smooth muscle cells. However, loss of Dgcr8 causes a significant portion of the cNCCs to undergo apoptosis, causing a decrease in the pool of progenitors required for OFT remodeling. Our data uncover a new role of Dgcr8 in cardiovascular morphogenesis, plausibly as part of transmission mechanism for FGF-dependent survival cue for migrating cNCCs
MicroRNA regulation of the paired-box transcription factor Pax3 confers robustness to developmental timing of myogenesis
No full textCommitment of progenitors in the dermomyotome to myoblast fate is the first step in establishing the body musculature. Pax3 is a crucial transcription factor, important for skeletal muscle development and expressed in myogenic progenitors in the dermomyotome of developing somites and in migratory muscle progenitors that populate the limb buds. Down-regulation of Pax3 is essential to ignite the myogenic program, including up-regulation of myogenic regulators, Myf-5 and MyoD. MicroRNAs (miRNAs) confer robustness to developmental timing by posttranscriptional repression of genetic programs that are related to previous developmental stages or to alternative cell fates. Here we demonstrate that the muscle-specific miRNAs miR-1 and miR-206 directly target Pax3. Antagomir-mediated inhibition of miR-1/miR-206 led to delayed myogenic differentiation in developing somites, as shown by transient loss of myogenin expression. This correlated with increased Pax3 and was phenocopied using Pax3-specific target protectors. Loss of myogenin after antagomir injection was rescued by Pax3 knockdown using a splice morpholino, suggesting that miR-1/miR-206 control somite myogenesis primarily through interactions with Pax3. Our studies reveal an important role for miR-1/miR-206 in providing precision to the timing of somite myogenesis. We propose that posttranscriptional control of Pax3 downstream of miR-1/miR-206 is required to stabilize myoblast commitment and subsequent differentiation. Given that mutually exclusive expression of miRNAs and their targets is a prevailing theme in development, our findings suggest that miRNA may provide a general mechanism for the unequivocal commitment underlying stem cell differentiation
Erythrocyte survival is controlled by microRNA-142
No full textHematopoietic–specific microRNA-142 is a critical regulator of various blood cell lineages, but its role in erythrocytes is unexplored. Herein, we characterize the impact of microRNA-142 on erythrocyte physiology and molecular cell biology, using a mouse loss-of-function allele. We report that microRNA-142 is required for maintaining the typical erythrocyte biconcave shape and structural resilience, for the normal metabolism of reactive oxygen species, and for overall lifespan. microRNA-142 further controls ACTIN filament homeostasis and membrane skeleton organization. The analyses presented reveal previously unappreciated functions of microRNA-142 and contribute to an emerging view of small RNAs as key players in erythropoiesis. Finally, the work herein demonstrates how a housekeeping network of cytoskeletal regulators can be reshaped by a single micro-RNA denominator in a cell type specific manner
miRNA malfunction causes spinal motor neuron disease
No full textDefective RNA metabolism is an emerging mechanism involved in ALS pathogenesis and possibly in other neurodegenerative disorders. Here, we show that microRNA (miRNA) activity is essential for long-term survival of postmitotic spinal motor neurons (SMNs) in vivo. Thus, mice that do not process miRNA in SMNs exhibit hallmarks of spinal muscular atrophy (SMA), including sclerosis of the spinal cord ventral horns, aberrant end plate architecture, and myofiber atrophy with signs of denervation. Furthermore, a neurofilament heavy subunit previously implicated in motor neuron degeneration is specifically up-regulated in miRNA-deficient SMNs. We demonstrate that the heavy neurofilament subunit is a target of miR-9, a miRNA that is specifically down-regulated in a genetic model of SMA. These data provide evidence for miRNA function in SMN diseases and emphasize the potential role of miR-9–based regulatory mechanisms in adult neurons and neurodegenerative states.</jats:p
Whole-genome sequencing reveals that variants in the Interleukin 18 Receptor Accessory Protein 3′UTR protect against ALS
No full textThe noncoding genome is substantially larger than the protein-coding genome but has been largely unexplored by genetic association studies. Here, we performed region-based rare variant association analysis of >25,000 variants in untranslated regions of 6,139 amyotrophic lateral sclerosis (ALS) whole genomes and the whole genomes of 70,403 non-ALS controls. We identified interleukin-18 receptor accessory protein (IL18RAP) 3' untranslated region (3'UTR) variants as significantly enriched in non-ALS genomes and associated with a fivefold reduced risk of developing ALS, and this was replicated in an independent cohort. These variants in the IL18RAP 3'UTR reduce mRNA stability and the binding of double-stranded RNA (dsRNA)-binding proteins. Finally, the variants of the IL18RAP 3'UTR confer a survival advantage for motor neurons because they dampen neurotoxicity of human induced pluripotent stem cell (iPSC)-derived microglia bearing an ALS-associated expansion in C9orf72, and this depends on NF-κB signaling. This study reveals genetic variants that protect against ALS by reducing neuroinflammation and emphasizes the importance of noncoding genetic association studies.sponsorship: We gratefully acknowledge the contributions of all participants and the investigators who provided biological samples and data for the Project Mine ALS sequencing consortium, the NYGC ALS Consortium, the gnomAD and TOPMed of the NHLBI (https://www.nhlbiwgs.org/topmed-banner-authorship).We thank M. Ward (NINDS, NIH) for sharing human inducible i3LMN cells. Samples used in this research were in part obtained from the UK National DNA Bank for MND Research, funded by the MND Association and the Wellcome Trust. We acknowledge sample management undertaken by Biobanking Solutions funded by the Medical Research Council at the Centre for Integrated Genomic Medical Research, University of Manchester. We would like to thank the NINDS Biorepository at Coriell Institute for iPSC cell lines used in this study. We thank B. Oldak and J. Hanna for microglia differentiation protocols, N. Kozer and H. Barr for assistance with live-cell imaging, A. Savidor and Y. Levin for mass spectrometry and M. Shmueli, Y. Merbl and R. Rotkof for advice and protocols. We thank LSE for language and scientific editing. Some illustrations were created with BioRender. The Hornstein lab is supported by friends of S. Brenner. E.H. is Head of Andi and Larry Wolfe Center for Research on Neuroimmunology and Neuromodulation and incumbent of Ira & Gail Mondry Professorial chair. This work is funded by Legacy Heritage Fund (828/17), Bruno and Ilse Frick Foundation for Research on ALS, the RADALA Foundation for ALS research, Teva Pharmaceutical Industries., Ltd., as part of the Israeli National Network of Excellence in Neuroscience (NNE) and Minna-James-Heineman Stiftung through Minerva, the European Research Council under the European Union's Seventh Framework Programme (FP7/2007-2013)/ERC grant agreement number 617351, Israel Science Foundation (135/16, 3497/21); Target ALS 118945, the Minerva Foundation, with funding from the Federal German Ministry for Education and Research, the ALS-Therapy Alliance, AFM Telethon (20576 to E.H.), Motor Neuron Disease Association (UK), The Thierry Latran Foundation for ALS research, ERA-Net for Research Programmes on Rare Diseases (FP7), via the Israel Ministry of Health. A. Alfred Taubman through IsrALS, Yeda-Sela, Yeda-CEO, Israel Ministry of Trade and Industry, Y. Leon Benoziyo Institute for Molecular Medicine, Kekst Family Institute for Medical Genetics, David and Fela Shapell Family Center for Genetic Disorders Research, Crown Human Genome Center, Nathan, Shirley, Philip and Charlene Vener New Scientist Fund, Julius and Ray Charlestein Foundation, Fraida Foundation, Wolfson Family Charitable Trust, Adelis Foundation, Merck (UK), Maria Halphen, Estates of Fannie Sherr, Lola Asseof, Lilly Fulop, Andi and Larry Wolfe Center for Research on Neuroimmunology and Neuromodulation and Benoziyo center for Neurological diseases, Weizmann-Brazil Center for Research on Neurodegeneration at The Weizmann Institute of Science, Redhill Foundation-Sam and Jean Rothberg Charitable Trust, Edward and Janie Moravitz, the Israeli Council for Higher Education via the Weizmann Data Science Research Center and a research grant from the Estate of Tully and Michele Plesser and M. Judith Ruth Institute for Preclinical Brain Research. A.A.-C.received funding from Neurodegenerative Disease Research (JPND), Medical Research Council (MR/L501529/1, STRENGTH, MR/R024804/1, BRAIN-MEND), Economic and Social Research Council (ES/L008238/1, ALS-CarE), MND Association, National Institute for Health Research (NIHR) Biomedical Research Centre at South London and Maudsley NHS Foundation Trust and King's College London. This project has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (grant agreement number 772376, EScORIAL). The collaboration project is cofunded by the PPP Allowance made available by Health~Holland, Top Sector Life Sciences & Health to stimulate public-private partnerships. This study was supported by the ALS Foundation Netherlands. For P.V.D., Project MinE Belgium was supported by a grant from IWT (number 140935), the ALS Liga Belgie, the National Lottery of Belgium and the KU Leuven Opening the Future Fund. P.V.D. holds a senior clinical investigatorship of FWO-Vlaanderen and is supported by E. von Behring Chair for Neuromuscular and Neurodegenerative Disorders, the ALS Liga Belgie and the KU Leuven funds `Een Hart voor ALS', `Laeversfonds voor ALS Onderzoek' and the `Valery Perrier Race against ALS Fund'. Several authors of this publication are members of the European Reference Network for Rare Neuromuscular Diseases. P.J.S. received funding from the Medical Research Council, MND Association, NIHR Senior Investigator Award, NIHR Sheffield Biomedical Research Centre and NIHR Sheffield Clinical Research Facility. P.M.A. received funding from the Knut and Alice Wallenberg Foundation, the Swedish Brain Foundation, the Swedish Science Council and the Ulla-Carin Lindquist Foundation. H.P.P. and sequencing activities at NYGC were supported by the ALS Association and The Tow Foundation. C.E. was supported by a scholarship from Teva Pharmaceutical Industries, Ltd., as part of the NNE. S.M.K.F. is supported by the ALS Canada Tim E. Noel Postdoctoral Fellowship. R.H.B.J. was funded by the ALS Association, ALS Finding a Cure, Angel Fund, ALS-One, Cellucci Fund and NIH grants (R01 NS104022, R01 NS073873 and NS111990-01 to R.H.B.J.). J.K.I. is a New York Stem Cell Foundation-Robertson Investigator. N.S.Y. was supported by the Israeli Council for Higher Education via the Weizmann Data Science Research Center, by a research grant from the Estate of Tully and Michele Plesser and by Maccabim Foundation. Work in the J.K.I. lab was supported by NIH grant R01NS097850, U.S. Department of Defense grant W81XWH-19-PRARP-CSRA and grants from the Tau Consortium, the New York Stem Cell Foundation, the ALS Association and the John Douglas French Alzheimer's Foundation. R.L.McL. received funding from the Science Foundation Ireland (17/CDA/4737), and A.N.B. received funding from the Suna and Inan Kirac Foundation. J.E.L. received funding from the National Institute of Health/NINDS (R01 NS073873). (MND Association, Wellcome Trust, Medical Research Council at the Centre for Integrated Genomic Medical Research, University of Manchester, Legacy Heritage Fund|828/17, Bruno and Ilse Frick Foundation for Research on ALS, RADALA Foundation for ALS research, Teva Pharmaceutical Industries., Ltd., as part of the Israeli National Network of Excellence in Neuroscience (NNE), Minna-James-Heineman Stiftung through Minerva, European Research Council under the European Union|617351, Israel Science Foundation|135/16, Israel Science Foundation|3497/21, Target ALS|118945, Minerva Foundation, Federal German Ministry for Education and Research, ALS-Therapy Alliance, AFM Telethon|20576, Motor Neuron Disease Association (UK), Thierry Latran Foundation for ALS research, ERA-Net for Research Programmes on Rare Diseases (FP7), via the Israel Ministry of Health, Estate of Tully and Michele Plesser, Neurodegenerative Disease Research (JPND), Medical Research Council|MR/L501529/1, Medical Research Council|MR/R024804/1, Economic and Social Research Council (ALS-CarE)|ES/L008238/1, National Institute for Health Research (NIHR) Biomedical Research Centre at South London and Maudsley NHS Foundation Trust and King's College London, European Research Council (ERC) under the European Union|772376, PPP Allowance, ALS Foundation Netherlands, IWT|140935, ALS Liga Belgie, National Lottery of Belgium, KU Leuven Opening the Future Fund, E. von Behring Chair for Neuromuscular and Neurodegenerative Disorders, KU Leuven funds 'Een Hart voor ALS', Laeversfonds voor ALS Onderzoek, Valery Perrier Race against ALS Fund, Medical Research Council, NIHR Senior Investigator Award, NIHR Sheffield Biomedical Research Centre, NIHR Sheffield Clinical Research Facility, Knut and Alice Wallenberg Foundation, Swedish Brain Foundation, Swedish Science Council, Ulla-Carin Lindquist Foundation, ALS Association, Tow Foundation, Teva Pharmaceutical Industries, Ltd., as part of the NNE, ALS Canada Tim E. Noel Postdoctoral Fellowship, ALS Finding a Cure, Angel Fund, ALS-One, NIH|R01 NS104022, NIH|R01 NS073873, NIH|NS111990-01, NIH|R01NS097850, Israeli Council for Higher Education via the Weizmann Data Science Research Center, Maccabim Foundation, U.S. Department of Defense|W81XWH-19-PRARP-CSRA, Tau Consortium, New York Stem Cell Foundation, John Douglas French Alzheimer's Foundation, Science Foundation Ireland|17/CDA/4737, Suna and Inan Kirac Foundation, National Institute of Health/NINDS|R01 NS073873, Cellucci Fund, ESRC|ES/L008238/1, MRC|MR/L501529/1, MRC|MR/R024804/1, European Research Council (ERC)|772376, National Institute of Neurological Disorders and Stroke|R01NS104022, National Institute on Aging; National Institute of Neurological Disorders and Stroke|R01NS097850, Economic and Social Research Council|ES/L008238/1, Medical Research Council|MR/L501529/1, Medical Research Council|MR/R024804/1, Motor Neurone Disease Association|AlChalabi-Dobson/Apr14/829-791, National Institute for Health Research|NIHR202421, National Institute for Health Research|NF-SI-0617-10077)status: Publishe
