PMID- 28334782
OWN - NLM
STAT- MEDLINE
DCOM- 20171017
LR  - 20231213
IS  - 1460-2083 (Electronic)
IS  - 0964-6906 (Linking)
VI  - 26
IP  - 9
DP  - 2017 May 1
TI  - Golgi-retained Cx32 mutants interfere with gene addition therapy for CMT1X.
PG  - 1622-1633
LID - 10.1093/hmg/ddx064 [doi]
AB  - Numerous GJB1 gene mutations cause the X-linked form of Charcot-Marie-Tooth 
      disease (CMT1X). GJB1 encodes connexin32 (Cx32), which forms trans-myelin gap 
      junctions in Schwann cells. Most GJB1 mutations result in loss-of-function 
      mechanisms, supporting the concept of gene replacement therapy. However, 
      interactions between delivered wild type and endogenously expressed mutant Cx32 
      may potentially occur in the setting of gene replacement therapy. In order to 
      screen for possible interactions of several representative CMT1X mutants with 
      wild type Cx32 that may interfere with the functional gap junction formation, we 
      established an in vitro screening method co-expressing in HeLa cells wild type 
      Cx32 and one of eight different Cx32 mutants including A39P, A39V, T55I, R75W, 
      M93V, L143P, N175D and R183S. Some of the Golgi-retained mutants hindered gap 
      junction plaque assembly by Cx32 on the cell membrane, while 
      co-immunoprecipitation analysis revealed a partial interaction of wild type 
      protein with Golgi-retained mutants. Dye transfer studies confirmed that 
      Golgi-retained R75W, M93V and N175D but not endoplasmic reticulum-retained T55I 
      had a negative effect on wild type Cx32 function. Finally, in vivo intraneural 
      delivery of the gene encoding the wild type Cx32 in mice bearing either the T55I 
      or R75W mutation on Cx32 knockout background showed that virally delivered 
      protein was correctly localized in mice expressing the endoplasmic 
      reticulum-retained T55I whereas it did not traffic normally in mice expressing 
      the Golgi-retained R75W. Thus, certain Golgi-retained Cx32 mutants may interfere 
      with exogenously delivered Cx32. Screening for mutant-wild type Cx32 interactions 
      should be considered prior to planning gene addition therapy for CMT1X.
CI  - (c) The Author 2017. Published by Oxford University Press. All rights reserved. For 
      Permissions, please email: journals.permissions@oup.com.
FAU - Kyriakoudi, Styliana
AU  - Kyriakoudi S
AD  - Neuroscience Laboratory.
FAU - Sargiannidou, Irene
AU  - Sargiannidou I
AD  - Neuroscience Laboratory.
FAU - Kagiava, Alexia
AU  - Kagiava A
AD  - Neuroscience Laboratory.
FAU - Olympiou, Margarita
AU  - Olympiou M
AD  - Neuroscience Laboratory.
FAU - Kleopa, Kleopas A
AU  - Kleopa KA
AD  - Neuroscience Laboratory.
AD  - Neurology Clinics, The Cyprus Institute of Neurology and Genetics and Cyprus 
      School of Molecular Medicine, 1683 Nicosia, Cyprus.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Hum Mol Genet
JT  - Human molecular genetics
JID - 9208958
RN  - 0 (Connexins)
SB  - IM
MH  - Animals
MH  - Charcot-Marie-Tooth Disease/*genetics/metabolism
MH  - Connexins/*genetics/*metabolism
MH  - Endoplasmic Reticulum/metabolism
MH  - Gap Junctions/metabolism
MH  - Genetic Therapy/methods
MH  - Golgi Apparatus/genetics/physiology
MH  - HeLa Cells/metabolism
MH  - Humans
MH  - Mice
MH  - Mice, Knockout
MH  - Mutation
MH  - Myelin Sheath/metabolism
MH  - Schwann Cells/metabolism
MH  - Gap Junction beta-1 Protein
EDAT- 2017/03/24 06:00
MHDA- 2017/10/19 06:00
CRDT- 2017/03/24 06:00
PHST- 2016/12/15 00:00 [received]
PHST- 2017/02/15 00:00 [accepted]
PHST- 2017/03/24 06:00 [pubmed]
PHST- 2017/10/19 06:00 [medline]
PHST- 2017/03/24 06:00 [entrez]
AID - 3039198 [pii]
AID - 10.1093/hmg/ddx064 [doi]
PST - ppublish
SO  - Hum Mol Genet. 2017 May 1;26(9):1622-1633. doi: 10.1093/hmg/ddx064.