Genetics, Gene Therapy & Genetic Disease
- Therapeutic gene editing in CD34+ hematopoietic progenitors from Fanconi anemia patients
- Begoña Diez1,2,3,
- Pietro Genovese4,
- Francisco J Roman‐Rodriguez1,2,3,
- Lara Alvarez1,2,3,
- Giulia Schiroli4,5,
- Laura Ugalde1,2,3,
- Sandra Rodriguez‐Perales6,
- Julian Sevilla3,7,8,
- Cristina Diaz de Heredia9,
- Michael C Holmes10,
- Angelo Lombardo4,5,
- Luigi Naldini4,5,
- Juan Antonio Bueren (juan.bueren{at}ciemat.es)*,1,2,3 and
- Paula Rio (paula.rio{at}ciemat.es)*,1,2,3
- 1Division of Hematopoietic Innovative Therapies, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas, Madrid, Spain
- 2Advanced Therapies Unit, Instituto de Investigación Sanitaria Fundación Jiménez Díaz, Madrid, Spain
- 3Centro de Investigación Biomédica en Red de Enfermedades Raras, Spain
- 4San Raffaele Telethon Institute for Gene Therapy (SR‐Tiget), IRCCS San Raffaele Scientific Institute, Milan, Italy
- 5Vita Salute San Raffaele University, Milan, Italy
- 6Molecular Cytogenetics Group, Human Cancer Genetics Program, Centro Nacional de Investigaciones Oncológicas (CNIO), Madrid, Spain
- 7Servicio Hemato‐Oncología Pediátrica, Hospital Infantil Universitario Niño Jesús, Madrid, Spain
- 8Fundación Investigación Biomédica, Hospital Infantil Universitario Niño Jesús, Madrid, Spain
- 9Servicio de Oncología y Hematología Pediátrica, Hospital Universitario Vall d'Hebron, Barcelona, Spain
- 10Sangamo Therapeutics, Inc., Richmond, CA, USA
- ↵*
Corresponding author. Tel: +34913466518; Fax: +34913466484; E‐mail: juan.bueren{at}ciemat.es
Corresponding author. Tel: +34913460890; Fax: +34913466484; E‐mail: paula.rio{at}ciemat.es
Zinc finger nuclease‐mediated gene editing facilitates insertion of a therapeutic gene in the AAVS1 “safe harbor” locus in human hematopoietic cell lines and primary HSPCs from healthy donors and patients with Fanconi anemia‐A, leading to their phenotypic correction.
Synopsis
Zinc finger nuclease‐mediated gene editing facilitates insertion of a therapeutic gene in the AAVS1 “safe harbor” locus in human hematopoietic cell lines and primary HSPCs from healthy donors and patients with Fanconi anemia‐A, leading to their phenotypic correction.
FANCA is not essential for zinc finger nuclease (ZFN)‐mediated gene editing in human hematopoietic cells.
ZFN‐mediated gene editing facilitates the targeting of FANCA in a “safe harbor” locus of lymphoblastic cell lines (LCLs) from Fanconi anemia A (FA‐A) patients.
ZFN‐mediated gene editing facilitates the targeting of donor constructs in long term hematopoietic stem and progenitor cells (HSPCs) from healthy donors.
Gene targeting of therapeutic constructs in HSPCs from FA‐A patients is thus feasible and mediates the phenotypic correction of these cells.
- Received January 15, 2017.
- Revision received August 9, 2017.
- Accepted August 14, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- An Alzheimer‐associated TREM2 variant occurs at the ADAM cleavage site and affects shedding and phagocytic function
- Kai Schlepckow1,
- Gernot Kleinberger1,2,
- Akio Fukumori3,7,
- Regina Feederle2,3,4,
- Stefan F Lichtenthaler2,3,5,6,
- Harald Steiner1,3 and
- Christian Haass (christian.haass{at}mail03.med.uni-muenchen.de)*,1,2,3
- 1Biomedical Center (BMC), Biochemistry, Ludwig‐Maximilians‐Universität München, Munich, Germany
- 2Munich Cluster for Systems Neurology (SyNergy), Munich, Germany
- 3German Center for Neurodegenerative Diseases (DZNE) Munich, Munich, Germany
- 4Helmholtz Center Munich, German Research Center for Environmental Health, Institute for Diabetes and Obesity, Core Facility Monoclonal Antibody Development, Neuherberg, Germany
- 5Neuroproteomics, Klinikum Rechts der Isar, Technische Universität München, Munich, Germany
- 6Institute for Advanced Study, Technische Universität München, Garching, Germany
- 7Present Address: Department of Psychiatry, Osaka University Health Care Center, Toyonaka Osaka, Japan
- ↵*Corresponding author. Tel: +49 89 4400 46549; E‐mail: christian.haass{at}mail03.med.uni-muenchen.de
The triggering receptor expressed on myeloid cells 2 (TREM2) is shed on the cell surface by ADAM10 and ADAM17 between histidine 157 and serine 158, a site where the AD associated p.H157Y variant was found. p.H157Y increases shedding and impairs phagocytic function by lowering cell surface TREM2.
Synopsis
The triggering receptor expressed on myeloid cells 2 (TREM2) is shed on the cell surface by ADAM10 and ADAM17 between histidine 157 and serine 158, a site where the AD associated p.H157Y variant was found. p.H157Y increases shedding and impairs phagocytic function by lowering cell surface TREM2.
The TREM2 ectodomain is cleaved by ADAM10/17 C‐terminal to histidine 157.
The late‐onset AD‐associated variant p.H157Y facilitates shedding and lowers cell surface mature TREM2.
Decreased mature p.H157Y TREM2 on the cell surface reduces phagocytic activity.
- Received February 7, 2017.
- Revision received June 26, 2017.
- Accepted July 14, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Genome editing for scalable production of alloantigen‐free lentiviral vectors for in vivo gene therapy
- Michela Milani1,2,
- Andrea Annoni1,
- Sara Bartolaccini1,
- Mauro Biffi1,
- Fabio Russo1,
- Tiziano Di Tomaso1,
- Andrea Raimondi3,
- Johannes Lengler4,
- Michael C Holmes5,
- Friedrich Scheiflinger4,
- Angelo Lombardo1,2,
- Alessio Cantore1,† and
- Luigi Naldini (luigi.naldini{at}hsr.it)*,1,2,†
- 1San Raffaele Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy
- 2Vita Salute San Raffaele University, Milan, Italy
- 3IRCCS San Raffaele Scientific Institute, Milan, Italy
- 4Baxalta (former Baxter) Innovation GmbH, Vienna, Austria
- 5Sangamo Therapeutics, Inc., Richmond, CA, USA
- ↵*Corresponding author. Tel: +39 02 2643 4681; E‐mail: luigi.naldini{at}hsr.it
Lentiviral vectors (LV) lacking major histocompatibility complexes (MHC) and with low vesicular stomatitis virus glycoprotein G (VSV.G) content escape immune recognition by human T cells and complement. Cell line packaged LV offer scalable and consistent LV production for in vivo gene therapy.
Synopsis
Lentiviral vectors (LV) lacking major histocompatibility complexes (MHC) and with low vesicular stomatitis virus glycoprotein G (VSV.G) content escape immune recognition by human T cells and complement. Cell line‐packaged LV offer scalable and consistent LV production for in vivo gene therapy.
Site‐specific integration of the LV genome into an inducible packaging cell line allows consistent generation of high‐yield producers of the LV of interest.
Cell line‐produced LV achieve equivalent gene transfer in the mouse liver as LV produced by conventional transfection but are more stable in human sera, due to a lower VSV.G content.
Genetic inactivation of beta‐2 microglobulin (B2M) in LV producer cells allows production of MHC‐free LV with preserved infectivity, devoid of immunogenic alloantigens in human recipients.
- Received June 13, 2017.
- Revision received July 21, 2017.
- Accepted July 26, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- One level up: abnormal proteolytic regulation of IGF activity plays a role in human pathophysiology
- Jesús Argente (jesus.argente{at}uam.es)*,1,2,3,4,
- Julie A Chowen1,3,
- Luis A Pérez‐Jurado5,6,7,
- Jan Frystyk8 and
- Claus Oxvig9
- 1Department of Pediatrics & Pediatric Endocrinology, Hospital Infantil Universitario Niño Jesús, Instituto de Investigación La Princesa, Madrid, Spain
- 2Department of Pediatrics, Universidad Autónoma de Madrid, Madrid, Spain
- 3Centro de Investigación Biomédica en Red de Fisiopatología de la Obesidad y Nutrición (CIBEROBN), Instituto de Salud Carlos III, Madrid, Spain
- 4IMDEA Food Institute, CEI UAM + CSIC, Madrid, Spain
- 5Genetics Unit, Universitat Pompeu Fabra, Barcelona, Spain
- 6Hospital del Mar Research Institute (IMIM), Barcelona, Spain
- 7Centro de Investigación Biomédica en Red de Enfermedades Raras (CIBERER), Instituto de Salud Carlos III, Barcelona, Spain
- 8Medical Research Laboratory, Department of Clinical Medicine, Health, Aarhus University, Aarhus C, Denmark
- 9Department of Molecular Biology & Genetics, Aarhus University, Aarhus C, Denmark
- ↵*Corresponding author. Tel: +34 915035912; Fax: +34 915035939; E‐mail: jesus.argente{at}uam.es
This review discusses how identifying patients’ mutations in new proteins can advance our understanding of the underlying pathophysiology, i.e. delayed growth failure, but also more widely, highlight new pathways of clinical relevance like the GH/IGF‐1 axis that offer real therapeutic opportunities.
- Received April 25, 2017.
- Revision received July 15, 2017.
- Accepted July 18, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- β‐Klotho sustains postnatal GnRH biology and spins the thread of puberty
- Micheline Misrahi (micheline.misrahi{at}aphp.fr)1
This article highlights the newly discovered role of β‐Klotho in postnatal GnRH biology through the identification of a patient congenital hypogonadotropic hypogonadism‐associated FGFR1 mutation that results in decreased FGF21 metabolic signaling.
- © 2017 The Author. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Clinical development of CAR T cells—challenges and opportunities in translating innovative treatment concepts
- Jessica Hartmann (jessica.hartmann{at}pei.de)*,1,
- Martina Schüßler‐Lenz2,3,
- Attilio Bondanza4 and
- Christian J Buchholz (christian.buchholz{at}pei.de)*,1,3
- 1Molecular Biotechnology and Gene Therapy, Paul‐Ehrlich‐Institut, Langen, Germany
- 2Division of Medical Biotechnology, Paul‐Ehrlich‐Institut, Langen, Germany
- 3German Cancer Consortium (DKTK), Heidelberg, Germany
- 4Innovative immunotherapies, Ospedale San Raffaele, Milano, Italy
- ↵*
Corresponding author. Tel: +49 6103774026; E‐mail: jessica.hartmann{at}pei.de
Corresponding author. Tel: +49 6103774011; E‐mail: christian.buchholz{at}pei.de
Authoritative, insightful overview of the problems and general hurdles preventing efficient clinical development of chimeric antigen receptor (CAR) T cell therapy for cancer as well as future opportunities.
EMBO Mol Med (2017) 9: 1183–1197
- Received December 19, 2016.
- Revision received July 3, 2017.
- Accepted July 11, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- KLB, encoding β‐Klotho, is mutated in patients with congenital hypogonadotropic hypogonadism
- Cheng Xu1,
- Andrea Messina1,
- Emmanuel Somm1,
- Hichem Miraoui1,18,
- Tarja Kinnunen2,
- James Acierno Jr1,
- Nicolas J Niederländer1,
- Justine Bouilly1,
- Andrew A Dwyer1,3,
- Yisrael Sidis1,
- Daniele Cassatella1,
- Gerasimos P Sykiotis1,
- Richard Quinton4,
- Christian De Geyter5,
- Mirjam Dirlewanger6,
- Valérie Schwitzgebel6,
- Trevor R Cole7,
- Andrew A Toogood8,
- Jeremy MW Kirk9,
- Lacey Plummer10,
- Urs Albrecht11,
- William F Crowley Jr10,
- Moosa Mohammadi12,
- Manuel Tena‐Sempere13,14,15,
- Vincent Prevot16,17 and
- Nelly Pitteloud (nelly.pitteloud{at}chuv.ch)*,1
- 1Service of Endocrinology, Diabetology & Metabolism, Lausanne University Hospital, Lausanne, Switzerland
- 2Department of Biology, School of Applied Sciences, University of Huddersfield, Huddersfield, UK
- 3University of Lausanne Institute of Higher Education and Research in Healthcare, Lausanne, Switzerland
- 4Institute for Genetic Medicine, University of Newcastle‐on‐Tyne, Newcastle‐on Tyne, UK
- 5Clinic of Gynecological Endocrinology and Reproductive Medicine, University Hospital, University of Basel, Basel, Switzerland
- 6Pediatric Endocrine and Diabetes Unit, Children's Hospital, University Hospitals and Faculty of Medicine, Geneva, Switzerland
- 7Department of Clinical Genetics, Birmingham Women's Hospital, Birmingham, UK
- 8Department of Endocrinology, Queen Elizabeth Hospital, University Hospitals Birmingham, Birmingham, UK
- 9Department of Endocrinology, Birmingham Children's Hospital, Birmingham, UK
- 10National Center for Translational Research in Reproduction and Infertility, Harvard Reproductive Endocrine Sciences Center of the Department of Medicine, Massachusetts General Hospital, Boston, MA, USA
- 11Department of Biology, Biochemistry, Faculty of Science, University of Fribourg, Fribourg, Switzerland
- 12Department of Biochemistry & Molecular Pharmacology, New York University School of Medicine, New York, NY, USA
- 13Department of Cell Biology, Physiology and Immunology, University of Cordoba, Cordoba, Spain
- 14Instituto Maimonides de Investigación Biomédica de Cordoba (IMIBIC/HURS), Cordoba, Spain
- 15CIBER Fisiopatología de la Obesidad y Nutrición, Instituto de Salud Carlos III, Cordoba, Spain
- 16Inserm, Laboratory of Development and Plasticity of the Neuroendocrine Brain, JPARC, Lille, France
- 17FHU 1000 Days for Health, School of Medicine, University of Lille, Lille, France
- 18Present Address: Division of Genetics, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA, USA
- ↵*Corresponding author. Tel: +41 021 314 87 96; E‐mail: nelly.pitteloud{at}chuv.ch
Defects in FGF21/KLB/FGFR1 signaling contribute to GnRH deficiency in both humans and mice. This signaling pathway is a novel link between metabolism and reproduction.
Synopsis
Defects in FGF21/KLB/FGFR1 signaling contribute to GnRH deficiency in both humans and mice. This signaling pathway is a novel link between metabolism and reproduction.
Heterozygous loss‐of‐function mutations in KLB are found in patients with congenital hypogonadotropic hypogonadism.
Klb‐deficient mice delayed sexual maturation and impaired fertility with decreased gonadotropins due to a hypothalamic defect.
Klb is expressed in the postnatal hypothalamus including GnRH neurons.
FGF21 reaches GnRH neurons via fenestrated capillaries in the hypothalamus in vivo and enhances GnRH release in median eminence explants in vitro.
- Received December 1, 2016.
- Revision received June 23, 2017.
- Accepted June 27, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Promoterless gene targeting without nucleases rescues lethality of a Crigler‐Najjar syndrome mouse model
- Fabiola Porro1,†,
- Giulia Bortolussi1,†,
- Adi Barzel2,3,
- Alessia De Caneva1,
- Alessandra Iaconcig1,
- Simone Vodret1,
- Lorena Zentilin1,
- Mark A Kay2 and
- Andrés F Muro (muro{at}icgeb.org)*,1
- 1International Centre for Genetic Engineering and Biotechnology (ICGEB), Trieste, Italy
- 2Departments of Pediatrics and Genetics, Stanford University, Stanford, CA, USA
- 3Present Address: Department of Biochemistry and Molecular Biology, The George S. Wise Faculty of Life Sciences, Tel Aviv University, Tel Aviv, Israel
- ↵*Corresponding author. Tel: +39 040 3757369; Fax: +39 040 226555; E‐mail: muro{at}icgeb.org
↵† These authors contributed equally to this work as first authors
Gene targeting into the albumin locus without using nucleases rescues lethality in a severe genetic liver disease in mice, thereby avoiding the loss of therapeutic DNA by hepatocyte duplication, a critical issue in AAV‐mediated gene replacement therapy.
Synopsis
Gene targeting into the albumin locus without using nucleases rescues lethality in a severe genetic liver disease in mice, thereby avoiding the loss of therapeutic DNA by hepatocyte duplication, a critical issue in AAV‐mediated gene replacement therapy.
The “GeneRide” approach applied to a severe Crigler‐Najjar mouse model fully rescues neonatal lethality, biochemical and functional abnormalities.
The therapy reduced plasma bilirubin to safe levels, resulting in normal motor‐coordination abilities.
The absence of nucleases and promoter‐driven transgene reduces most safety concerns of AAV‐mediated gene replacement therapy.
Our results show that the “GeneRide” approach is also effective for cytoplasmic and ER membrane‐bound proteins.
This proof‐of‐principle demonstrates the applicability of the approach to other genetic diseases of the liver.
- Received January 24, 2017.
- Revision received June 27, 2017.
- Accepted July 4, 2017.
- © 2017 International Centre for Genetic Engineering and Biotechnology Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- MG132‐induced progerin clearance is mediated by autophagy activation and splicing regulation
- Karim Harhouri1,
- Claire Navarro1,
- Danielle Depetris1,
- Marie‐Geneviève Mattei1,
- Xavier Nissan2,
- Pierre Cau1,3,
- Annachiara De Sandre‐Giovannoli1,4 and
- Nicolas Lévy (nicolas.levy{at}univ-amu.fr)*,1,4
- 1Aix Marseille Univ, INSERM, GMGF (Génétique Médicale et Génomique Fonctionnelle), Marseille, France
- 2CECS, I‐STEM, Institut des cellules Souches pour le Traitement et l'Etude des maladies Monogéniques, AFM, Evry, France
- 3AP‐HM, Hôpital la Timone, Service de Biologie Cellulaire, Marseille, France
- 4AP‐HM, Hôpital la Timone, Département de Génétique Médicale, Marseille, France
- ↵*Corresponding author. Tel: +33 4 91 32 48 97; Fax: +33 4 91 43 29 90; E‐mail: nicolas.levy{at}univ-amu.fr
Progerin is a toxic protein that accumulates in the nuclei of Progeria patients' cells, sequestered in abnormal PML‐NBs. The proteasome inhibitor MG132 is shown to degrade progerin by activating autophagy and transcriptional inhibition through SRSF‐1 and SRSF‐5 splicing regulation.
Synopsis
Progerin is a toxic protein that accumulates in the nuclei of Progeria patients' cells, sequestered in abnormal PML‐NBs. The proteasome inhibitor MG132 is shown to degrade progerin by activating autophagy and transcriptional inhibition through SRSF‐1 and SRSF‐5 splicing regulation.
Ubiquitinylated progerin is sequestered into abnormal ProMyelocytic Leukemia Nuclear Bodies (PML‐NBs).
Progerin reduction is based on MG132 dual action: autophagy activation and splicing regulation.
MG132 in vitro treatment rescues most of the biological hallmarks of progeria.
MG132 local treatment efficiently reduces progerin levels in vivo, in the LmnaG609G/G609G mouse model.
The powerful and dual activities of MG132 make it a promising drug towards a future and safe therapeutic development for Progeria and related Prelamin‐A processing defective diseases.
EMBO Mol Med (2017) 9: 1294–1313
- Received November 8, 2016.
- Revision received June 2, 2017.
- Accepted June 7, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
- Lentiviral vectors escape innate sensing but trigger p53 in human hematopoietic stem and progenitor cells
- Francesco Piras1,2,
- Michela Riba3,
- Carolina Petrillo1,2,
- Dejan Lazarevic3,
- Ivan Cuccovillo1,
- Sara Bartolaccini1,
- Elia Stupka3,4,
- Bernhard Gentner1,
- Davide Cittaro3,
- Luigi Naldini1,2 and
- Anna Kajaste‐Rudnitski (kajaste.anna{at}hsr.it)*,1
- 1San Raffaele Telethon Institute for Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy
- 2Vita‐Salute San Raffaele University, Milan, Italy
- 3Center for Translational Genomics and Bioinformatics, IRCCS San Raffaele Scientific Institute, Milan, Italy
- 4Present Address: Boehringer Ingelheim, Biberach an der Riß, Germany
- ↵*Corresponding author. Tel: +39 0226435007; E‐mail: kajaste.anna{at}hsr.it
Lentiviral (LV) gene therapy vectors escape innate sensing but trigger p53 signaling in human hematopoietic stem and progenitor cells (HSPC), ultimately leading to lower engraftment of short‐term repopulating cells.
Synopsis
Lentiviral (LV) gene therapy vectors escape innate sensing but trigger p53 signaling in human hematopoietic stem and progenitor cells (HSPC), ultimately leading to lower engraftment of short‐term repopulating cells.
LV transduction remains remarkably stealth in human HSPC, avoiding innate immune activation but triggering p53 signaling in human HSPC.
p53 signaling occurs upon ATM‐dependent nuclear sensing of vector DNA (LV, IDLV, AAV) independently of integration into the host genome.
Vector‐mediated activation of p53 leads to cell cycle arrest and apoptosis ex vivo ultimately leading to lower engraftment of short‐term HSPC in vivo.
Inhibition of LV‐signaling rescues ex vivo apoptosis and early engraftment of human HSPC.
EMBO Mol Med (2017) 9: 1198–1211
- Received April 21, 2017.
- Revision received June 6, 2017.
- Accepted June 7, 2017.
- © 2017 The Authors. Published under the terms of the CC BY 4.0 license
This is an open access article under the terms of the Creative Commons Attribution 4.0 License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
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