scholarly journals A novel BCR-ABL fusion gene (e6a2) in a patient with Philadelphia chromosome-negative chronic myelogenous leukemia

Blood ◽  
1996 ◽  
Vol 88 (6) ◽  
pp. 2236-2240 ◽  
Author(s):  
A Hochhaus ◽  
A Reiter ◽  
H Skladny ◽  
JV Melo ◽  
C Sick ◽  
...  
Keyword(s):  
Southern Blot ◽  
Chronic Myelogenous Leukemia ◽  
Rare Variants ◽  
Fusion Gene ◽  
Philadelphia Chromosome ◽  
Myelogenous Leukemia ◽  
Rt Pcr ◽  
Novel Variant ◽  
Polymerase Chain

A novel variant of the chimeric BCR-ABL mRNA transcript was detected in a patient with Philadelphia chromosome-negative (Ph-) chronic myelogenous leukemia (CML) by multiplex reverse-transcription polymerase chain reaction (RT-PCR). Sequence analysis of the fusion region of the amplified cDNA fragment showed an in-frame joining of exon e6 of the BCR gene and exon a2 of the ABL gene, giving rise to an e6a2 BCR-ABL transcript. This finding was confirmed by Southern blot analysis using a specific probe corresponding to intron 6 of the BCR gene, whereas conventional Southern blot for rearrangement of the major breakpoint cluster region (M-bcr) was negative. Western blot studies detected a BCR-ABL protein slightly larger than p185 BCR-ABL. Metaphase fluorescence in situ hybridization showed an insertion of ABL material into the BCR region without reciprocal BCR translocation. The findings in this case show that atypical BCR-ABL transcripts are detectable even in Ph- CML patients without M-bcr-rearrangements. Multiplex PCR using primers that allow for amplification of all known BCR-ABL transcripts is an appropriate method to exclude these rare variants.

scholarly journals Detection of two alternative bcr/abl mRNA junctions and minimal residual disease in Philadelphia chromosome positive chronic myelogenous leukemia by polymerase chain reaction

Blood ◽  
1989 ◽  
Vol 73 (8) ◽  
pp. 2165-2170
Author(s):  
MS Lee ◽  
A LeMaistre ◽  
HM Kantarjian ◽  
M Talpaz ◽  
EJ Freireich ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Myelogenous Leukemia ◽  
Philadelphia Chromosome ◽  
Rare Event ◽  
Chronic Phase ◽  
Myelogenous Leukemia ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Minimal Residual

The Philadelphia (Ph′) chromosome in chronic myelogenous leukemia (CML) results in fusion of the bcr gene and c-abl oncogene, which transcribes into two types of chimeric bcr/abl mRNAs: the L-6 junction and the K-28 junction. By means of a highly sensitive assay, combination of reverse transcription and polymerase chain reaction (RT/PCR), we analyzed 38 blood samples obtained from 31 patients with Ph′-positive CML and two patients with Ph′-negative bcr rearranged CML. Among the 21 samples obtained in chronic phase, eight patients had the L-6 mRNA, 11 had the K-28 mRNA, and two had both the L-6 and K-28 mRNAs. Among the nine samples obtained in blast crisis, four contained the L-6 mRNA, two contained the K-28 mRNA, and three contained both the K-28 and L-6 mRNAs. This finding supports the concept of alternative splicing of bcr/abl mRNAs transcribed in Ph′-positive CML. However, it appears to be a rare event. Of the eight samples obtained from eight patients who had achieved complete cytogenetic remission and negativity for bcr region rearrangement for 6 months to 3 years after recombinant alpha interferon (r alpha-IFN) therapy, all of them showed evidence of minimal residual Ph′-positive clones as detected by the RT/PCR assay. This finding suggests that interferon therapy suppresses the proliferation of the Ph′-positive clones, but it does not completely eradicate the Ph′-positive stem cells.

scholarly journals Detection of two alternative bcr/abl mRNA junctions and minimal residual disease in Philadelphia chromosome positive chronic myelogenous leukemia by polymerase chain reaction

Blood ◽  
1989 ◽  
Vol 73 (8) ◽  
pp. 2165-2170 ◽  
Author(s):  
MS Lee ◽  
A LeMaistre ◽  
HM Kantarjian ◽  
M Talpaz ◽  
EJ Freireich ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Myelogenous Leukemia ◽  
Philadelphia Chromosome ◽  
Rare Event ◽  
Chronic Phase ◽  
Myelogenous Leukemia ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Minimal Residual

Abstract The Philadelphia (Ph′) chromosome in chronic myelogenous leukemia (CML) results in fusion of the bcr gene and c-abl oncogene, which transcribes into two types of chimeric bcr/abl mRNAs: the L-6 junction and the K-28 junction. By means of a highly sensitive assay, combination of reverse transcription and polymerase chain reaction (RT/PCR), we analyzed 38 blood samples obtained from 31 patients with Ph′-positive CML and two patients with Ph′-negative bcr rearranged CML. Among the 21 samples obtained in chronic phase, eight patients had the L-6 mRNA, 11 had the K-28 mRNA, and two had both the L-6 and K-28 mRNAs. Among the nine samples obtained in blast crisis, four contained the L-6 mRNA, two contained the K-28 mRNA, and three contained both the K-28 and L-6 mRNAs. This finding supports the concept of alternative splicing of bcr/abl mRNAs transcribed in Ph′-positive CML. However, it appears to be a rare event. Of the eight samples obtained from eight patients who had achieved complete cytogenetic remission and negativity for bcr region rearrangement for 6 months to 3 years after recombinant alpha interferon (r alpha-IFN) therapy, all of them showed evidence of minimal residual Ph′-positive clones as detected by the RT/PCR assay. This finding suggests that interferon therapy suppresses the proliferation of the Ph′-positive clones, but it does not completely eradicate the Ph′-positive stem cells.

Identification of human chronic myelogenous leukemia progenitor cells with hemangioblastic characteristics

Blood ◽  
2005 ◽  
Vol 105 (7) ◽  
pp. 2733-2740 ◽  
Author(s):  
Baijun Fang ◽  
Chunmei Zheng ◽  
Lianming Liao ◽  
Qin Han ◽  
Zhao Sun ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endothelial Cells ◽  
Progenitor Cells ◽  
Chronic Myelogenous Leukemia ◽  
Blood Cells ◽  
Fusion Gene ◽  
Fetal Liver ◽  
Philadelphia Chromosome ◽  
Myelogenous Leukemia ◽  
Leukemic Cells

AbstractOverwhelming evidence from leukemia research has shown that the clonal population of neoplastic cells exhibits marked heterogeneity with respect to proliferation and differentiation. There are rare stem cells within the leukemic population that possess extensive proliferation and self-renewal capacity not found in the majority of the leukemic cells. These leukemic stem cells are necessary and sufficient to maintain the leukemia. Interestingly, the BCR/ABL fusion gene, which is present in chronic myelogenous leukemia (CML), was also detected in the endothelial cells of patients with CML, suggesting that CML might originate from hemangioblastic progenitor cells that can give rise to both blood cells and endothelial cells. Here we isolated fetal liver kinase-1–positive (Flk1+) cells carrying the BCR/ABL fusion gene from the bone marrow of 17 Philadelphia chromosome–positive (Ph+) patients with CML and found that these cells could differentiate into malignant blood cells and phenotypically defined endothelial cells at the single-cell level. These findings provide direct evidence for the first time that rearrangement of the BCR/ABL gene might happen at or even before the level of hemangioblastic progenitor cells, thus resulting in detection of the BCR/ABL fusion gene in both blood and endothelial cells.

scholarly journals Detection by enzymatic amplification of bcr-abl mRNA in peripheral blood and bone marrow cells of patients with chronic myelogenous leukemia

Blood ◽  
1989 ◽  
Vol 73 (6) ◽  
pp. 1735-1741 ◽  
Author(s):  
W Lange ◽  
DS Snyder ◽  
R Castro ◽  
JJ Rossi ◽  
KG Blume
Keyword(s):  
Bone Marrow ◽  
Chronic Myelogenous Leukemia ◽  
Peripheral Blood ◽  
Bone Marrow Cells ◽  
Philadelphia Chromosome ◽  
Chromosome 9 ◽  
Myelogenous Leukemia ◽  
Enzymatic Amplification ◽  
Polymerase Chain ◽  
Marrow Cells

Abstract The Philadelphia chromosome of chronic myelogenous leukemia (CML) patients is caused by a translocation of the c-abl gene from chromosome 9 to the breakpoint cluster region (bcr) on chromosome 22. A new bcr- abl mRNA is expressed in these cases. We have developed a modified polymerase chain reaction (PCR) for the detection of this mRNA. The method is extremely sensitive, reliable, and relatively fast. The analysis of peripheral blood or bone marrow cells from CML patients treated with chemotherapy shows that the two possible mRNAs are expressed in various combinations. Our results show that even after myeloablative therapy for bone marrow transplantation bcr-abl mRNAs are still expressed. Further studies, however, are necessary to determine the clinical relevance of a small number of persisting cells expressing the bcr-abl mRNA.

scholarly journals Multi-unit ribozyme-mediated cleavage of bcr-abl mRNA in myeloid leukemias

Blood ◽  
1995 ◽  
Vol 85 (8) ◽  
pp. 2162-2170 ◽  
Author(s):  
LH Leopold ◽  
SK Shore ◽  
TA Newkirk ◽  
RM Reddy ◽  
EP Reddy
Keyword(s):  
Chronic Myelogenous Leukemia ◽  
Fusion Gene ◽  
Folate Receptor ◽  
Philadelphia Chromosome ◽  
Chromosome 9 ◽  
Myelogenous Leukemia ◽  
Myeloid Leukemias ◽  
Transforming Activity ◽  
Cleavage Reactions

Chronic myelogenous leukemia is characterized by the Philadelphia chromosome, which at the molecular level results from the fusion of the bcr gene on chromosome 22 and the abl gene on chromosome 9. The bcr-abl fusion gene encodes a novel tyrosine kinase with transforming activity. In this study, we have synthesized a multi-unti ribozyme that targets bcr-abl mRNA. In vitro ribozyme cleavage reactions show increased cleavage efficiency of this multi-unit ribozyme compared with single or double ribozymes. The multiunit ribozyme was then transfected into murine myeloblasts transformed with the bcr-abl gene (32D cells). Ribozyme transfection was accomplished either by liposomes or using follic acid-polylysine as a carrier. Multi-unit ribozyme transfection reduced the level of bcr-abl mRNA 3 logs when transfected via folate receptor-mediated uptake into transformed 32D cells. These results suggest that a multi-unit ribozyme could be an effective therapeutic agent for the treatment of Philadelphia chromosome-positive chronic myelogenous leukemia.

scholarly journals Application of chromosome microdissection probes for elucidation of BCR- ABL fusion and variant Philadelphia chromosome translocations in chronic myelogenous leukemia

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3365-3371 ◽  
Author(s):  
J Zhang ◽  
P Meltzer ◽  
R Jenkins ◽  
XY Guan ◽  
J Trent
Keyword(s):  
Chronic Myelogenous Leukemia ◽  
Chromosome Rearrangement ◽  
Philadelphia Chromosome ◽  
Leukemic Cell ◽  
Myelogenous Leukemia ◽  
Chromosome Microdissection ◽  
Chromosome Translocations ◽  
Ph Chromosome

Abstract Fluorescence in situ hybridization (FISH) has become an increasingly important method for assessing chromosome rearrangement. The reciprocal translocation constituting the Philadelphia (Ph) chromosome [t(9;22)(q34;q11)] characterizes more than 90% of patients with chronic myelogenous leukemia (CML). However, in the remaining cases the Ph chromosome (genetically characterized by the fusion of the BCR-ABL genes) is thought to arise through complex translocations that are often not readily apparent using routine chromosome-banding analysis. For this reason we have developed unique band-specific probes for two- color FISH that detect unequivocally the Ph chromosome, and its derivatives. Results of the application of these probes are illustrated by analysis of 11 cases of CML (9 of which contain “variant” translocations). The probes were generated by chromosome microdissection and in vitro amplification of the bands involved in the Ph translocation, leading to an extremely fast and sensitive approach to identify this alteration in leukemic cell populations.

scholarly journals BCR-ABL rearrangements in children with Philadelphia chromosome- positive chronic myelogenous leukemia

Blood ◽  
1991 ◽  
Vol 78 (9) ◽  
pp. 2407-2410
Author(s):  
I Aurer ◽  
A Butturini ◽  
RP Gale
Keyword(s):  
Chronic Myelogenous Leukemia ◽  
Southern Blotting ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
Myelogenous Leukemia ◽  
Chain Reaction ◽  
Molecular Rearrangement ◽  
Exon 2 ◽  
Exon 1 ◽  
Polymerase Chain

Leukemia cells from adults with Philadelphia (Ph1)-chromosome positive chronic myelogenous leukemia (CML) have a characteristic molecular rearrangement between the BCR and ABL genes whereby major breakpoint cluster region (Mbcr) exons 2 or 3 are joined to ABL exon II. Ph1- chromosome positive CML is uncommon in children and it is unknown whether these children have similar rearrangements. We studied 17 children with Ph1-chromosome positive CML. Five were studied for Mbcr rearrangement using Southern blotting, nine for the presence of chimeric BCR-ABL mRNA using reverse transcription and polymerase chain reaction, and three for both. All eight children studied by Southern blotting had BCR rearrangement. Of 12 children in whom BCR-ABL mRNA was studied, 10 had Mbcr exon 2 joined to ABL exon II, one had Mbcr exon 3 joined to ABL II, and one had both Mbcr-ABL junctions. These data indicate a similarity to adult CML. However, mRNA processing in children may preferentially splice Mbcr exon 2 to ABL exon II. No child had BCR exon 1 joined to ABL exon II, the rearrangement typical of childhood Ph1-chromosome positive acute lymphoblastic leukemia.

scholarly journals Nanostring-based screening for tyrosine kinase fusions in inflammatory myofibroblastic tumors

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Taisei Kurihara ◽  
Yoshiyuki Suehara ◽  
Keisuke Akaike ◽  
Takuo Hayashi ◽  
Shinji Kohsaka ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Rna Sequencing ◽  
Fusion Gene ◽  
Rt Pcr ◽  
Anaplastic Lymphoma ◽  
Inflammatory Myofibroblastic Tumors ◽  
Polymerase Chain ◽  
Lymphoplasmacytic Infiltration ◽  
Tyrosine Receptor Kinase

Abstract Gene expression imbalances were measured for tyrosine kinase (TK) genes using Nanostring in 19 samples of inflammatory myofibroblastic tumor (IMT). All cases were immunohistochemically stained with anaplastic lymphoma kinase (ALK) and pan-tropomyosin-related-kinase (pan-Trk) antibodies. Five cases with imbalanced ALK expression, reported with Nanostring, were tested using fluorescence in situ hybridization (FISH); two cases with imbalanced neurotrophic tyrosine receptor kinase 3 (NTRK3) expression were tested using reverse transcription-polymerase chain reaction (RT-PCR). One case with imbalanced expression for ROS proto-oncogene 1 (ROS1) was tested using RNA sequencing and RT-PCR. TK fusions were detected in all cases with imbalanced TK expression. RNA sequencing detected a FN1–ROS1 fusion gene in an adult IMT case. IMT with ALK rearrangement showed myofibroblast-dominant features. IMT with ETV6–NTRK3 fusion showed prominent lymphoplasmacytic infiltration with scattered myofibroblasts. Pan-Trk IHC revealed only scattered positively stained cells in IMT with ETV6–NTRK3 fusion gene. ROS1-positive IMT showed myofibroblast-dominant features.

scholarly journals Application of chromosome microdissection probes for elucidation of BCR- ABL fusion and variant Philadelphia chromosome translocations in chronic myelogenous leukemia

Blood ◽  
1993 ◽  
Vol 81 (12) ◽  
pp. 3365-3371
Author(s):  
J Zhang ◽  
P Meltzer ◽  
R Jenkins ◽  
XY Guan ◽  
J Trent
Keyword(s):  
Chronic Myelogenous Leukemia ◽  
Chromosome Rearrangement ◽  
Philadelphia Chromosome ◽  
Leukemic Cell ◽  
Myelogenous Leukemia ◽  
Chromosome Microdissection ◽  
Chromosome Translocations ◽  
Ph Chromosome

Fluorescence in situ hybridization (FISH) has become an increasingly important method for assessing chromosome rearrangement. The reciprocal translocation constituting the Philadelphia (Ph) chromosome [t(9;22)(q34;q11)] characterizes more than 90% of patients with chronic myelogenous leukemia (CML). However, in the remaining cases the Ph chromosome (genetically characterized by the fusion of the BCR-ABL genes) is thought to arise through complex translocations that are often not readily apparent using routine chromosome-banding analysis. For this reason we have developed unique band-specific probes for two- color FISH that detect unequivocally the Ph chromosome, and its derivatives. Results of the application of these probes are illustrated by analysis of 11 cases of CML (9 of which contain “variant” translocations). The probes were generated by chromosome microdissection and in vitro amplification of the bands involved in the Ph translocation, leading to an extremely fast and sensitive approach to identify this alteration in leukemic cell populations.

Signicance of Philadelphia Chromosome in Chronic Myeloid Leukemia Patients of Anmol Hospital, Lahore, Pakistan

2018 ◽  
Vol 1 (1) ◽  
Author(s):  
Kanwal Rashid ◽  
Amber Hassan ◽  
Imrana Tanvir ◽  
Kashifa Ehsan
Keyword(s):  
Chronic Myelogenous Leukemia ◽  
Myeloid Leukemia ◽  
Philadelphia Chromosome ◽  
Experimental Studies ◽  
Chromosome 8 ◽  
Myelogenous Leukemia ◽  
Chromosomal Marker ◽  
Rt Pcr ◽  
Ph Chromosome ◽  
Cell Disorder

Chronic myelogenous Leukemia is (Clonal stem cell disorder) cancer which starts in bone marrow a soft tissue inside bones that aids to form blood cells. It is rst form of cancer that was rstly recognize to associate strongly with the chromosomal abnormality and the chromosome [t (9; 22) translocation] called Philadelphia chromosome. Abstract: Objective: Philadelphia chromosome is a characteristic chromosomal marker that is associated with chronic myelogenous leukemia. Methods: More than one hundred patients of either sex were selected for the experiment. RNA was isolated from whole blood of patients so can use exclusively in RT-PCR. Results: Philadelphia chromosome in blood samples of patients with suspected diagnosis of CML was detected in 63% of patients. During our experimental studies on CML patients we do not encounter any complex translocation involving chromosome 8, 9 and 22. Conclusions: Philadelphia chromosome is a precise cytogenetic marker the detection of which is signicant for differential diagnosis and clinical organization of patients with clinical diagnosis of CML. It is of signicant that Ph chromosome occurs in pre-leukemic stage and has great diagnostic signicance.

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