PMID- 12558785
OWN - NLM
STAT- MEDLINE
DCOM- 20030623
LR  - 20220223
IS  - 1351-8216 (Print)
IS  - 1351-8216 (Linking)
VI  - 9
IP  - 1
DP  - 2003 Jan
TI  - Bone marrow mesenchymal cells for haemophilia A gene therapy using retroviral 
      vectors with modified long-terminal repeats.
PG  - 94-103
AB  - Bone marrow (BM) cells are attractive target cells for ex vivo gene therapy of 
      genetic diseases, including haemophilia A. However, BM-derived haematopoietic 
      stem/progenitor cells (HSCs) transduced with factor VIII (FVIII) retroviral 
      vectors, failed to express FVIII in vivo. To overcome the limitations of HSCs for 
      haemophilia gene therapy, BM-derived mesenchymal cells were explored as 
      alternative target cells. The BM mesenchymal cell population contains 
      self-renewing mesenchymal stem/progenitor cells that give rise to different 
      mesenchymal lineages and have been used safely in phase I gene-marking trials. 
      Human BM mesenchymal cells were transduced in vitro with an improved retroviral 
      vector encoding a human B-domain deleted FVIII (hFVIIIdeltaB) cDNA 
      (MND-MFG-hFVIIIdeltaB). This vector contains multiple modifications in the 
      cis-acting elements within the MoMLV long-terminal repeats (LTR) that prevent the 
      binding of repressive transcription factors. These modifications were previously 
      shown to increase and prolong gene expression in embryonic stem (ES) cells and 
      HSCs. Transduction of BM mesenchymal cells with the MND-MFG-hFVIIIdeltaB 
      retroviral vector resulted in high levels of functional human FVIII in vitro, 
      ranging between 300 +/- 50 SD and 700 +/- 100 SD mU per 106 cells per 24 h. 
      Following xenografting of the transduced human BM cells into immunodeficient 
      NOD-SCID mice, therapeutic hFVIII levels of 12 +/- 10 ng mL-1 were detected in 
      the plasma. Polymerase chain reaction analysis demonstrated long-term engraftment 
      (>3 months) of the human BM mesenchymal cells. The long-term persistence of BM 
      mesenchymal cells in the absence of myelo-ablative conditioning and the 
      therapeutic FVIII levels in vivo underscore the potential usefulness of 
      BM-derived mesenchymal cells for haemophilia gene therapy, as opposed to 
      BM-derived HSCs. Despite the modifications of the MoMLV LTR, FVIII expression 
      declined, which coincided with a decrease in FVIII mRNA transcription levels, 
      indicating that the salutary effect of the LTR modification on transgene 
      expression is not universally applicable to all cell types.
FAU - Van Damme, A
AU  - Van Damme A
AD  - Center for Transgene Technology and Gene Therapy, Flanders Interuniversity 
      Institute for Biotechnology, University of Leuven, Capus UZ Gasthuisberg, Leuven, 
      Belgium.
FAU - Chuah, M K L
AU  - Chuah MK
FAU - Dell'accio, F
AU  - Dell'accio F
FAU - De Bari, C
AU  - De Bari C
FAU - Luyten, F
AU  - Luyten F
FAU - Collen, D
AU  - Collen D
FAU - VandenDriessche, T
AU  - VandenDriessche T
LA  - eng
PT  - Journal Article
PL  - England
TA  - Haemophilia
JT  - Haemophilia : the official journal of the World Federation of Hemophilia
JID - 9442916
RN  - 9001-27-8 (Factor VIII)
SB  - IM
EIN - Haemophilia. 2003 May;9(3):345
MH  - Animals
MH  - Bone Marrow Transplantation/*methods
MH  - Factor VIII/biosynthesis/*genetics
MH  - Gene Transfer Techniques
MH  - Genetic Therapy/*methods
MH  - Genetic Vectors
MH  - Hemophilia A/*therapy
MH  - Humans
MH  - Mice
MH  - Mice, Inbred NOD
MH  - Mice, SCID
MH  - Polymerase Chain Reaction/methods
MH  - Retroviridae/genetics
MH  - Reverse Transcriptase Polymerase Chain Reaction
MH  - Stromal Cells/transplantation
MH  - Terminal Repeat Sequences/*genetics
MH  - Transplantation, Heterologous
EDAT- 2003/02/01 04:00
MHDA- 2003/06/24 05:00
CRDT- 2003/02/01 04:00
PHST- 2003/02/01 04:00 [pubmed]
PHST- 2003/06/24 05:00 [medline]
PHST- 2003/02/01 04:00 [entrez]
AID - 709 [pii]
AID - 10.1046/j.1365-2516.2003.00709.x [doi]
PST - ppublish
SO  - Haemophilia. 2003 Jan;9(1):94-103. doi: 10.1046/j.1365-2516.2003.00709.x.