PMID- 16952195
OWN - NLM
STAT- MEDLINE
DCOM- 20070131
LR  - 20151119
IS  - 1099-498X (Print)
IS  - 1099-498X (Linking)
VI  - 8
IP  - 10
DP  - 2006 Oct
TI  - Gene therapy for BCR-ABL+ human CML with dual phosphorylation resistant p27Kip1 
      and stable RNA interference using an EBV vector.
PG  - 1251-61
AB  - BACKGROUND: BCR-ABL-mediated chronic myelogenous leukemia (CML) CD34(+) cell 
      proliferation mostly depends on the nucleo-cytoplasmic ratio of the 
      cyclin-dependent kinase inhibitor p27. The ubiquitin-ligase SCF(Skp2) promotes 
      degradation of phosphorylated p27 at T187 in the nucleus, resulting in G1/S 
      progression of the cells. On the other hand, phosphatidylinositol-3-kinase 
      (PI3K)-directed T157 nuclear localization signal (NLS) phosphorylation results in 
      cytoplasmic sequestration of p27, leading to abnormal integrin-mediated 
      proliferation of CD34(+) CML cells. METHODS: We demonstrate the generation of an 
      engineered Epstein-Barr virus (EBV) vector with a BAC backbone that has the 
      unique capacity to carry doubly modified (DM) p27 (i.e. T187A, T157A p27) along 
      with the BCR-ABL siRNA expression construct. The HSV-tk suicide gene has also 
      been incorporated in the same vector, which promotes apoptosis in a 
      BCR-ABL-independent pathway. RESULTS: Expression of DM p27 markedly inhibits 
      proliferation of BCR-ABL(+) primary human CML cells. Moreover, DM p27 strongly 
      inhibits the growth of imatinib-resistant CML cells, compared to the T157A p27 
      (SM p27). The CML growth inhibition is found to be the result of significant G1/S 
      arrest with concomitant increase in hypophosphorylated retinoblastoma (Rb). 
      Moreover, the EBV vector mediated stable RNA interference induces apoptosis in 
      K562 cells and reduces myeloid colony forming units. CONCLUSIONS: We therefore 
      propose a multi-gene delivery strategy for BCR-ABL(+) CML cells by targeting not 
      only the fusion transcript, but also the downstream signaling, to overcome drug 
      resistance in the acute phase of CML.
CI  - Copyright (c) 2006 John Wiley & Sons, Ltd.
FAU - Sengupta, Amitava
AU  - Sengupta A
AD  - Structural Genomics Section & Biophysics Division, Saha Institute of Nuclear 
      Physics, Kolkata 700 064, India.
FAU - Banerjee, Debasish
AU  - Banerjee D
FAU - Chandra, Sarmila
AU  - Chandra S
FAU - Banerjee, Subrata
AU  - Banerjee S
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Gene Med
JT  - The journal of gene medicine
JID - 9815764
RN  - 0 (Benzamides)
RN  - 0 (Piperazines)
RN  - 0 (Pyrimidines)
RN  - 0 (RNA, Small Interfering)
RN  - 147604-94-2 (Cyclin-Dependent Kinase Inhibitor p27)
RN  - 8A1O1M485B (Imatinib Mesylate)
RN  - EC 2.7.10.2 (Fusion Proteins, bcr-abl)
SB  - IM
MH  - Active Transport, Cell Nucleus
MH  - Apoptosis/drug effects
MH  - Benzamides
MH  - Cell Proliferation/drug effects
MH  - Cyclin-Dependent Kinase Inhibitor p27/*genetics/metabolism
MH  - Fusion Proteins, bcr-abl/*antagonists & inhibitors
MH  - G1 Phase
MH  - Genes, Transgenic, Suicide
MH  - Genetic Engineering
MH  - Genetic Therapy/*methods
MH  - Genetic Vectors/*chemical synthesis
MH  - Herpesvirus 4, Human/*genetics
MH  - Humans
MH  - Imatinib Mesylate
MH  - K562 Cells
MH  - Leukemia, Myelogenous, Chronic, BCR-ABL Positive/*therapy
MH  - Models, Biological
MH  - Phosphorylation
MH  - Piperazines/pharmacology
MH  - Point Mutation
MH  - Pyrimidines/pharmacology
MH  - RNA Interference
MH  - RNA, Small Interfering/*chemical synthesis
MH  - Retinoblastoma/metabolism
MH  - S Phase
MH  - Tumor Cells, Cultured
EDAT- 2006/09/05 09:00
MHDA- 2007/02/01 09:00
CRDT- 2006/09/05 09:00
PHST- 2006/09/05 09:00 [pubmed]
PHST- 2007/02/01 09:00 [medline]
PHST- 2006/09/05 09:00 [entrez]
AID - 10.1002/jgm.959 [doi]
PST - ppublish
SO  - J Gene Med. 2006 Oct;8(10):1251-61. doi: 10.1002/jgm.959.