PMID- 8750625
OWN - NLM
STAT- MEDLINE
DCOM- 19961017
LR  - 20190116
IS  - 1042-8194 (Print)
IS  - 1026-8022 (Linking)
VI  - 20
IP  - 1-2
DP  - 1995 Dec
TI  - Effects of BCR-ABL antisense oligonucleotides (AS-ODN) on human chronic myeloid 
      leukemic cells: AS-ODN as effective purging agents.
PG  - 67-76
AB  - We examined phosphorothioate oligodeoxyribonucleotides (ODNs) directed against 
      bcr in exon 3 or exon 2, which are rearranged with exon 2 of abl (B3A2 and B2A2) 
      at t(9;22) of chronic myelogenous leukemia (CML). Since these ODNs are designed 
      to be CML cell specific, we studied their effects on the human CML cell line 
      K562, which is known to have B3A2 rearrangement, and leukemic cells from 
      patients, as well as normal hematopoietic stem cells in vitro. In vitro 
      experiments were performed to determine a potential role of these two ODNs as ex 
      vivo purging agents. Incubation of B3A2 antisense at 40, 80, and 120 
      micrograms/ml with K562 CML cells for 72 hours at 37 degrees C resulted in 44%, 
      56%, and 63% reduction of CFU-L as compared to controls. In contrast, B3A2 sense 
      and B2A2 antisense had no significant growth inhibitory effect on K562 cells. 
      Incubation of B3A2 and B2A2 antisense ODNs at concentration of 80 micrograms/ml 
      at 37 degrees C for 36 hours with normal peripheral blood stem/progenitor cells 
      (PBSC) resulted in 124% and 98% CFU-GM formation as compared to untreated 
      controls, respectively. However, incubation of PBSC with B3A2 and B2A2 sense-ODNs 
      resulted in a 22% and 44% reduction in CFU-GM, respectively. In order to 
      determine the ex vivo purging effects of bcr-abl ODNs, the K562 cells were mixed 
      with PBSC from normal donors at a ratio of 1:20 (CML:PBSC). The mixture of cells 
      was then incubated with B3A2 antisense at 80 micrograms/ml for 36 hrs at 37 
      degrees C. After incubation, no CML cells were detected by fluorescence in situ 
      hybridization (FISH) as compared to untreated controls. These results were 
      confirmed by RT-PCR using bcr-abl primers and mRNA isolated from the mixture of 
      cells. Further, these results support the hypothesis that bcr-abl antisense ODNs 
      are potentially effective agents for ex vivo purging of autologous stem cells 
      before transplantation to eliminate/reduce the burden of leukemic cells. No 
      significant toxicity to normal hematopoietic stem/progenitor cell population by 
      the bcr-abl antisense ODNs was observed. Although unanticipated reductions in 
      normal hematopoietic progenitor cells (CFU-GM) were observed with sense ODNs, no 
      reduction in CFU-GM was observed with unrelated phosphorothioate ODN controls.
FAU - Wu, A G
AU  - Wu AG
AD  - Department of Cell Biology and Anatomy, University of Nebraska Medical Center, 
      Omaha 68198-3330, USA.
FAU - Joshi, S S
AU  - Joshi SS
FAU - Chan, W C
AU  - Chan WC
FAU - Iversen, P L
AU  - Iversen PL
FAU - Jackson, J D
AU  - Jackson JD
FAU - Kessinger, A
AU  - Kessinger A
FAU - Pirruccello, S J
AU  - Pirruccello SJ
FAU - Sanger, W G
AU  - Sanger WG
FAU - Sharp, J G
AU  - Sharp JG
FAU - Verbik, D J
AU  - Verbik DJ
AU  - et al.
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Leuk Lymphoma
JT  - Leukemia & lymphoma
JID - 9007422
RN  - 0 (DNA Primers)
RN  - 0 (Oligonucleotides, Antisense)
RN  - 0 (RNA, Messenger)
RN  - 0 (Thionucleotides)
RN  - EC 2.7.10.2 (Fusion Proteins, bcr-abl)
SB  - IM
MH  - Base Sequence
MH  - Bone Marrow Purging/*methods
MH  - Cell Division/drug effects
MH  - Cell Line
MH  - Colony-Forming Units Assay
MH  - DNA Primers
MH  - Exons
MH  - Fusion Proteins, bcr-abl/*biosynthesis/*genetics
MH  - Gene Expression
MH  - Hematopoietic Stem Cells/cytology/*drug effects
MH  - Humans
MH  - In Situ Hybridization, Fluorescence
MH  - Leukemia, Erythroblastic, Acute
MH  - Leukemia, Myelogenous, Chronic, BCR-ABL Positive
MH  - Molecular Sequence Data
MH  - Oligonucleotides, Antisense/*pharmacology
MH  - Polymerase Chain Reaction
MH  - RNA, Messenger/analysis/biosynthesis
MH  - Thionucleotides
MH  - Tumor Cells, Cultured
EDAT- 1995/12/01 00:00
MHDA- 1995/12/01 00:01
CRDT- 1995/12/01 00:00
PHST- 1995/12/01 00:00 [pubmed]
PHST- 1995/12/01 00:01 [medline]
PHST- 1995/12/01 00:00 [entrez]
AID - 10.3109/10428199509054755 [doi]
PST - ppublish
SO  - Leuk Lymphoma. 1995 Dec;20(1-2):67-76. doi: 10.3109/10428199509054755.