Advanced

Lentiviral gene transfer of TCIRG1 into peripheral blood CD34(+) cells restores osteoclast function in infantile malignant osteopetrosis.

Moscatelli, Ilana LU ; Thudium, Christian LU ; Flores Bjurström, Carmen LU ; Schulz, Ansgar ; Askmyr, Maria LU ; Gudmann, Natasja Stæhr ; Andersen, Nanna Merete ; Porras, Oscar ; Karsdal, Morten Asser and Villa, Anna , et al. (2013) In Bone 57(1). p.1-9
Abstract
Infantile malignant osteopetrosis (IMO) is a rare, lethal, autosomal recessive disorder characterized by non-functional osteoclasts. More than 50% of the patients have mutations in the TCIRG1 gene, encoding for a subunit of the osteoclast proton pump. The aim of this study was to restore the resorptive function of IMO osteoclasts by lentiviral mediated gene transfer of the TCIRG1 cDNA. CD34(+) cells from peripheral blood of five IMO patients and from normal cord blood were transduced with lentiviral vectors expressing TCIRG1 and GFP under a SFFV promoter, expanded in culture and differentiated on bone slices to mature osteoclasts. qPCR analysis and western blot revealed increased mRNA and protein levels of TCIRG1, comparable to controls.... (More)
Infantile malignant osteopetrosis (IMO) is a rare, lethal, autosomal recessive disorder characterized by non-functional osteoclasts. More than 50% of the patients have mutations in the TCIRG1 gene, encoding for a subunit of the osteoclast proton pump. The aim of this study was to restore the resorptive function of IMO osteoclasts by lentiviral mediated gene transfer of the TCIRG1 cDNA. CD34(+) cells from peripheral blood of five IMO patients and from normal cord blood were transduced with lentiviral vectors expressing TCIRG1 and GFP under a SFFV promoter, expanded in culture and differentiated on bone slices to mature osteoclasts. qPCR analysis and western blot revealed increased mRNA and protein levels of TCIRG1, comparable to controls. Vector corrected IMO osteoclasts generated increased release of Ca(2+) and bone degradation product CTX-I into the media as well as increased formation of resorption pits in the bone slices, while non-corrected IMO osteoclasts failed to resorb bone. Resorption was approximately 70-80% of that of osteoclasts generated from cord blood. Furthermore, transduced CD34(+) cells successfully engrafted in NSG-mice. In conclusion we provide the first evidence of lentiviral-mediated correction of a human genetic disease affecting the osteoclastic lineage. (Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; ; ; ; and , et al. (More)
; ; ; ; ; ; ; ; ; ; ; and (Less)
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Bone
volume
57
issue
1
pages
1 - 9
publisher
Elsevier
external identifiers
  • wos:000325742600001
  • pmid:23907031
  • scopus:84882696526
  • pmid:23907031
ISSN
1873-2763
DOI
10.1016/j.bone.2013.07.026
language
English
LU publication?
yes
id
29536bb8-2365-4b13-ba75-b6d564adcf48 (old id 4006272)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/23907031?dopt=Abstract
date added to LUP
2016-04-01 10:20:42
date last changed
2020-01-29 01:33:36
@article{29536bb8-2365-4b13-ba75-b6d564adcf48,
  abstract     = {Infantile malignant osteopetrosis (IMO) is a rare, lethal, autosomal recessive disorder characterized by non-functional osteoclasts. More than 50% of the patients have mutations in the TCIRG1 gene, encoding for a subunit of the osteoclast proton pump. The aim of this study was to restore the resorptive function of IMO osteoclasts by lentiviral mediated gene transfer of the TCIRG1 cDNA. CD34(+) cells from peripheral blood of five IMO patients and from normal cord blood were transduced with lentiviral vectors expressing TCIRG1 and GFP under a SFFV promoter, expanded in culture and differentiated on bone slices to mature osteoclasts. qPCR analysis and western blot revealed increased mRNA and protein levels of TCIRG1, comparable to controls. Vector corrected IMO osteoclasts generated increased release of Ca(2+) and bone degradation product CTX-I into the media as well as increased formation of resorption pits in the bone slices, while non-corrected IMO osteoclasts failed to resorb bone. Resorption was approximately 70-80% of that of osteoclasts generated from cord blood. Furthermore, transduced CD34(+) cells successfully engrafted in NSG-mice. In conclusion we provide the first evidence of lentiviral-mediated correction of a human genetic disease affecting the osteoclastic lineage.},
  author       = {Moscatelli, Ilana and Thudium, Christian and Flores Bjurström, Carmen and Schulz, Ansgar and Askmyr, Maria and Gudmann, Natasja Stæhr and Andersen, Nanna Merete and Porras, Oscar and Karsdal, Morten Asser and Villa, Anna and Fasth, Anders and Henriksen, Kim and Richter, Johan},
  issn         = {1873-2763},
  language     = {eng},
  number       = {1},
  pages        = {1--9},
  publisher    = {Elsevier},
  series       = {Bone},
  title        = {Lentiviral gene transfer of TCIRG1 into peripheral blood CD34(+) cells restores osteoclast function in infantile malignant osteopetrosis.},
  url          = {http://dx.doi.org/10.1016/j.bone.2013.07.026},
  doi          = {10.1016/j.bone.2013.07.026},
  volume       = {57},
  year         = {2013},
}