title: A novel disorder reveals clathrin heavy chain-22 is essential for human pain and touch development.
abstract: Congenital inability to feel pain is very rare but the identification of causative genes has yielded significant insights into pain pathways and also novel targets for pain treatment. We report a novel recessive disorder characterized by congenital insensitivity to pain, inability to feel touch, and cognitive delay. Affected individuals harboured a homozygous missense mutation in CLTCL1 encoding the CHC22 clathrin heavy chain, p.E330K, which we demonstrate to have a functional effect on the protein. We found that CLTCL1 is significantly upregulated in the developing human brain, displaying an expression pattern suggestive of an early neurodevelopmental role. Guided by the disease phenotype, we investigated the role of CHC22 in two human neural crest differentiation systems; human induced pluripotent stem cell-derived nociceptors and TRKB-dependant SH-SY5Y cells. In both there was a significant downregulation of CHC22 upon the onset of neural differentiation. Furthermore, knockdown of CHC22 induced neurite outgrowth in neural precursor cells, which was rescued by stable overexpression of small interfering RNA-resistant CHC22, but not by mutant CHC22. Similarly, overexpression of wild-type, but not mutant, CHC22 blocked neurite outgrowth in cells treated with retinoic acid. These results reveal an essential and non-redundant role for CHC22 in neural crest development and in the genesis of pain and touch sensing neurons.
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK msn27@cam.ac.uk cw347@cam.ac.uk.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Nahorski
authors.affiliation: 2 Department of Paediatrics, College of Medicine and Health Sciences, United Arab Emirates University, Al-Ain, UAE.
authors.forename: Lihadh
authors.initials: L
authors.lastname: Al-Gazali
authors.affiliation: 3 Department of Paediatrics, Tawam Hospital, Al-Ain, UAE.
authors.forename: Jozef
authors.initials: J
authors.lastname: Hertecant
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK.
authors.forename: David J
authors.initials: DJ
authors.lastname: Owen
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK 4 Max Planck Institute of Biochemistry, Department of Proteomics and Signal Transduction, Am Klopferspitz 18, 82152 Martinsried, Germany.
authors.forename: Georg H H
authors.initials: GH
authors.lastname: Borner
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK.
authors.forename: Ya-Chun
authors.initials: YC
authors.lastname: Chen
authors.affiliation: 5 Neusentis, The Portway Building, Granta Park, Cambridge. CB21 6GS, UK.
authors.forename: Caroline L
authors.initials: CL
authors.lastname: Benn
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK.
authors.forename: Ofélia P
authors.initials: OP
authors.lastname: Carvalho
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK.
authors.forename: Samiha S
authors.initials: SS
authors.lastname: Shaikh
authors.affiliation: 5 Neusentis, The Portway Building, Granta Park, Cambridge. CB21 6GS, UK.
authors.forename: Anne
authors.initials: A
authors.lastname: Phelan
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK.
authors.forename: Margaret S
authors.initials: MS
authors.lastname: Robinson
authors.affiliation: 6 Division of Biomedical Cell Biology, Warwick Medical School, Gibbet Hill Road, Coventry CV4 7AL, UK.
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Royle
authors.affiliation: 1 Cambridge Institute for Medical Research, University of Cambridge, Cambridge, CB2 0XY, UK msn27@cam.ac.uk cw347@cam.ac.uk.
authors.forename: C Geoffrey
authors.initials: CG
authors.lastname: Woods
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D033922:Clathrin Heavy Chains / Q000235:genetics*; D006801:Humans; D018482:Muscle, Skeletal / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D058953:Neural Stem Cells / Q000166:cytology*; D055495:Neurogenesis / Q000502:physiology*; D009474:Neurons / Q000378:metabolism; D010146:Pain / Q000235:genetics* / Q000378:metabolism; D014110:Touch / Q000502:physiology*
chemical_list: C101272:CLTCL1 protein, human; D033922:Clathrin Heavy Chains
keywords: clathrin; endosomal trafficking; insensitivity to pain; neurogenesis