title: A brain-sparing diphtheria toxin for chemical genetic ablation of peripheral cell lineages.
abstract: Conditional expression of diphtheria toxin receptor (DTR) is widely used for tissue-specific ablation of cells. However, diphtheria toxin (DT) crosses the blood-brain barrier, which limits its utility for ablating peripheral cells using Cre drivers that are also expressed in the central nervous system (CNS). Here we report the development of a brain-sparing DT, termed BRAINSPAReDT, for tissue-specific genetic ablation of cells outside the CNS. We prevent blood-brain barrier passage of DT through PEGylation, which polarizes the molecule and increases its size. We validate BRAINSPAReDT with regional genetic sympathectomy: BRAINSPAReDT ablates peripheral but not central catecholaminergic neurons, thus avoiding the Parkinson-like phenotype associated with full dopaminergic depletion. Regional sympathectomy compromises adipose tissue thermogenesis, and renders mice susceptible to obesity. We provide a proof of principle that BRAINSPAReDT can be used for Cre/DTR tissue-specific ablation outside the brain using CNS drivers, while consolidating the link between adiposity and the sympathetic nervous system.
authors.affiliation: Obesity Laboratory, Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal.
authors.forename: Mafalda M A
authors.initials: MM
authors.lastname: Pereira
authors.affiliation: Obesity Laboratory, Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal.
authors.forename: Inês
authors.initials: I
authors.lastname: Mahú
authors.affiliation: Obesity Laboratory, Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal.
authors.forename: Elsa
authors.initials: E
authors.lastname: Seixas
authors.affiliation: NeurObesity Group, Department of Physiology, CIMUS, University of Santiago de Compostela-Instituto de Investigación Sanitaria, Santiago de Compostela (A Coruña) 15782, Spain.|CIBER Fisiopatologia de la Obesidad y Nutrición (CIBERobn), Santiago de Compostela 15706, Spain.
authors.forename: Noelia
authors.initials: N
authors.lastname: Martinéz-Sánchez
authors.affiliation: Obesity Laboratory, Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal.
authors.forename: Nadiya
authors.initials: N
authors.lastname: Kubasova
authors.affiliation: Obesity Laboratory, Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal.
authors.forename: Roksana M
authors.initials: RM
authors.lastname: Pirzgalska
authors.affiliation: Laboratory of Molecular Metabolism, The Rockefeller University, New York, New York 10065, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Cohen
authors.affiliation: Section of Comparative Medicine, Yale University School of Medicine, New Haven, Connecticut 06520, USA.|Department of Neuroscience, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
authors.forename: Marcelo O
authors.initials: MO
authors.lastname: Dietrich
authors.affiliation: NeurObesity Group, Department of Physiology, CIMUS, University of Santiago de Compostela-Instituto de Investigación Sanitaria, Santiago de Compostela (A Coruña) 15782, Spain.|CIBER Fisiopatologia de la Obesidad y Nutrición (CIBERobn), Santiago de Compostela 15706, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: López
authors.affiliation: Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, UK.|Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Lisboa 1649-028, Portugal.
authors.forename: Gonçalo J L
authors.initials: GJ
authors.lastname: Bernardes
authors.affiliation: Obesity Laboratory, Instituto Gulbenkian de Ciência, Oeiras 2780-156, Portugal.
authors.forename: Ana I
authors.initials: AI
authors.lastname: Domingos
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology*; D019070:Cell Lineage; D004167:Diphtheria Toxin / Q000737:chemistry / Q000378:metabolism*; D004198:Disease Susceptibility; D004734:Energy Metabolism; D005247:Feeding Behavior; D005260:Female; D017353:Gene Deletion; D005786:Gene Expression Regulation; D018149:Glucose Intolerance / Q000150:complications / Q000473:pathology; D006367:HeLa Cells; D006801:Humans; D057446:Hydrodynamics; D008297:Male; D051379:Mice; D009068:Movement; D009474:Neurons / Q000378:metabolism; D009638:Norepinephrine / Q000378:metabolism; D009765:Obesity / Q000150:complications / Q000473:pathology; D010300:Parkinson Disease / Q000473:pathology / Q000503:physiopathology; D011092:Polyethylene Glycols / Q000737:chemistry; D013562:Sympathectomy; D022722:Thermogenesis / Q000235:genetics
chemical_list: D004167:Diphtheria Toxin; D011092:Polyethylene Glycols; D009638:Norepinephrine