Detection of the breakpoint cluster region-ABL fusion in chronic myeloid leukemia with variant Philadelphia chromosome translocations by in situ hybridization

1996 ◽  
Vol 89 (2) ◽  
pp. 153-156 ◽  
Author(s):  
Sabrina Tosi ◽  
Georges Cabot ◽  
Giovanni Giudici ◽  
Viviana Attuati ◽  
Paolo Morandi ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chronic Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Philadelphia Chromosome ◽  
Breakpoint Cluster Region ◽  
Chromosome Translocations ◽  
Cluster Region

scholarly journals توصيف حالة جديدة للانتقال الكروموسومي المتغاير t(9;22) في ابيضاض الدم النخاعي المزمن CMLبوساطة التهجين الموضعي التألقي FISH

2017 ◽  
Vol 1 (28) ◽  
pp. 609-620
Author(s):  
ظافر حسن غالي ◽  
انتصار حسين احمد
Keyword(s):  
In Situ Hybridization ◽  
Chronic Myeloid Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Myeloid Leukemia ◽  
Derivative Chromosome ◽  
Breakpoint Cluster Region ◽  
Cluster Region ◽  
Murine Leukemia

يعد ابيضاض الدم النخاعي المزمن" Chronic Myeloid Leukemia (CML) "ورم نخاعي تكاثري ويصيب 1 -2 شخص لكل 100000 بالغ. الامراضية الرئيسة للمرض هي تكوين اندماج جيني للجين Abelson murine leukemia (ABL1)" "على كروموسوم 9 مع الجين" breakpoint cluster region (BCR)"على كروموسوم 22.وهذا يؤدي الى التعبير عن بروتين ورمي يدعى "BCR-ABL1"  .في هذه الدراسة، سجلت حالة جديدة  متغايرة لابيضاض الدم النخاعي المزمن تحمل الانتقال الكروموسومي فلادلفيا""Philadelphia(Ph) translocation الذي تضمن الكروموسومين 9 و22 فضلا عن كروموسوم 3.انجز التوصيف الوراثي الخلوي الجزيئي بتقنية التهجين الموضعي التألقي "fluorescence in situ hybridization (FISH)", الذي اظهر وجودالاندماج الجيني 5'BCR/3'ABL1 على كروموسوم فلادلفيا ،في حين لم يشخص الاندماج الجيني 5'ABL1/3'BCR على مشتقة كروموسوم 9 derivative chromosome بسبب انتقال3'BCR الى الكروموسوم المشارك رقم 3.تقدم  هذه الدراسة تحليل وراثي خلوي جزيئي دقيق لحالة جديدة متغايرة من ابيضاض الدم النخاعي المزمن المتضمن كروموسوم فلادلفيا المعقد t(9;22).وتؤكد الدراسة ايضا دور تحليل التهجين الموضعي التألقي في تفسير هذه الانتقالات الكروموسومية التي هي خارج نطاق الوراثة الخلوية التقليدية.

scholarly journals Detection of chimeric BCR-ABL genes on bone marrow samples and blood smears in chronic myeloid and acute lymphoblastic leukemia by in situ hybridization

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1922-1928 ◽  
Author(s):  
M Bentz ◽  
G Cabot ◽  
M Moos ◽  
MR Speicher ◽  
A Ganser ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Acute Lymphoblastic Leukemia ◽  
Yeast Artificial Chromosome ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
High Specificity ◽  
Breakpoint Cluster Region ◽  
Cluster Region ◽  
Blood Smears

Abstract The presence of BCR-ABL fusion genes has important diagnostic and prognostic implications in chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL). The CML-specific chimeric BCR-ABL gene with a break involving the major breakpoint cluster region (M-bcr) of the BCR-gene has been detected by means of fluorescence in situ hybridization (FISH). In this study, we present a FISH protocol that allows the detection of breaks in both the major and the minor breakpoint cluster region (m-bcr). Three hybridization signals of D107F9, a yeast artificial chromosome (YAC)-derived probe spanning the breakpoint regions of the BCR gene, were indicative of the translocation events. To increase the specificity further, this probe was combined with cos-abl 8, a cosmid probe flanking the breakpoint within the ABL gene for dual-color hybridization. Samples of 21 patients with CML, the ALL-derived cell line SUP-B15, and of seven patients with Philadelphia chromosome (Ph1)-positive ALL (three of them with breakpoints within m-bcr) were examined. BCR-ABL fusion was detected in all cases with high specificity (false-positive nuclei: mean, 0.1%). On cytogenetic preparations, the percentages of BCR-ABL- positive interphase cells ranged from 53% to 91%. Comparable efficiencies were achieved on blood smears. In conclusion, hybridization with D107F9 and cos-abl 8 allows unambiguous diagnosis of BCR-ABL genes and is likely to become an important tool for the monitoring of therapies in patients with CML and ALL.

scholarly journals Aberrant methylation of the major breakpoint cluster region in chronic myeloid leukemia

Blood ◽  
1996 ◽  
Vol 88 (6) ◽  
pp. 2241-2249 ◽  
Author(s):  
CE Litz ◽  
JA Vos ◽  
CM Copenhaver
Keyword(s):  
Sequence Analysis ◽  
Chronic Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Philadelphia Chromosome ◽  
Nucleotide Position ◽  
Aberrant Methylation ◽  
Breakpoint Cluster Region ◽  
Cluster Region ◽  
Breakpoint Location

Isolated hypomethylated sites exist in the major breakpoint cluster region (M-bcr) where most Philadelphia chromosome (Ph) breakpoints are located. Twenty of 50 (40%) chronic myeloid leukemia (CML) patients were found to have aberrant hypermethylation of these sites on the rearranged M-bcr when compared with control marrows. The aberrancy correlated strongly with M-bcr breakpoint location; 19 of 20 cases had breakpoints located 5′ of the M-bcr Sca I site, and 28 of 30 cases with normal M-bcr methylation had breakpoints located 3′ of the M-bcr Sca I site. Sequence analysis of the Ph M-bcr breakpoints failed to find an M- bcr nucleotide position that delineated the transition between abnormally and normally methylated cases, indicating that the translocation of a critical M-bcr sequence was not responsible for the methylation abnormality. In 3 of 8 CML patients, cells without the t(9;22) were found to have abnormally methylated, unrearranged M-bcrs. The data indicate that abnormally methylated rearranged M-bcrs are present in CML cases with Ph breakpoints 5′ of the M-bcr Sca I site and that the M-bcr in Ph- cells of patients with CML may also be abnormally methylated.

A Novel t(9;22;11) Translocation Variant Involving 11q23 in a Patient with Chronic Myeloid Leukemia

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 7-7
Author(s):  
Lin Yuehui ◽  
Qinglong Zheng ◽  
Tong Wu ◽  
DAN LIU
Keyword(s):  
Flow Cytometry ◽  
In Situ Hybridization ◽  
Chronic Myeloid Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Myeloid Leukemia ◽  
Conflicts Of Interest ◽  
Fusion Gene ◽  
Philadelphia Chromosome ◽  
Flow Cytometry Analysis

Background:The progression of Philadelphia chromosome positive chronic myeloid leukemia (CML) is frequently accompanied by cytogenetic evolution, commonly unbalanced chromosomal changes. but balanced chromosomal translocations are very rare in CML, especially translocations involving the 11q23. The few reported cases with blast phase (BP) of CML carrying a 11q23 rearrangement results in insufficient responses to tyrosine kinase inhibitors (TKIs) and possess a poor prognosis. Methods:Cytogenetic analysis , fluorescence in situ hybridization (FISH), RNA sequencing (RNA-seq), targeted genomic sequencing and multi-parametric flow cytometry analysis were performed to identify the chromosome translocations and pathogenic gene alterations in a 36-year-old female with myeloid BP of CML. Results: In BP, the bone marrow (BM) aspiration showed 61% myeloid blasts; Multi-parametric flow cytometry analysis revealed the abnormal myeloid blasts expression of the following antigens: CD117, CD13, CD33, CD38, partially expressed CD15, CD64. Chromosome analysis revealed a t(11;22)(q23;q11) translocation in addition to the t(9;22)(q34;q11). Fluorescence in situ hybridization (FISH) test confirmed that the t(11;22)(q23;q11) involved the mixed lineage leukemia(MLL) gene on 11q23 and RNA sequence revealed MLL-SEPT5 and BCR/ABL1(p210) fusion transcripts positive. Mutations on 339 commonly mutated genes in hematologic malignancies were analyzed by targeted next-generation sequencing showed ASXL1 p.G949Vfs*2 mutation. The patient failed to respond to both imatinib and dasatinib despite the absence of resistance-associated mutations in the BCR/ABL1 gene and she had a myeloid blast crisis at 19 months after initiation of first- and second-generation TKI treatment. After BP, she received ponatinb, a third-generation TKI with chemotherapy. Regretly she didn't achieve a complete remission(CR) and was in the process of salvage transplantation at present. Conclusions:The presence of 11q23 rearrangements in BC of CML is rare and most likely accounts for the adverse clinical outcome. We first report a patient who diagnosed CML with t(11;22)(q23;q11) and MLL-SEPT5 fusion gene positive in BP of CML. The clinical course was aggressive, and therapy was poorly tolerated. Disclosures No relevant conflicts of interest to declare.

Highly Sensitive Fluorescence In Situ Hybridization Method to Detect Double BCR/ABL Fusion and Monitor Response to Therapy in Chronic Myeloid Leukemia

Blood ◽  
1998 ◽  
Vol 91 (9) ◽  
pp. 3357-3365 ◽  
Author(s):  
Gordon W. Dewald ◽  
William A. Wyatt ◽  
Amy L. Juneau ◽  
Richard O. Carlson ◽  
Alan R. Zinsmeister ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Chronic Myeloid Leukemia ◽  
Fluorescence In Situ Hybridization ◽  
Myeloid Leukemia ◽  
Philadelphia Chromosome ◽  
Response To Therapy ◽  
Chromosome 9 ◽  
Ph Chromosome ◽  
Cytogenetic Remission

Abstract We investigated a new method using fluorescence in situ hybridization and DNA probes that span the common breakpoints of t(9;22)(q34;q11.2) and that detect double BCR/ABL fusion (D-FISH) in bone marrow cells with this translocation, one on the abnormal chromosome 9 and one on the Philadelphia chromosome (Ph chromosome). D-FISH patterns were abnormal in 30 of 30 specimens with classic, simple, complex, and masked Ph chromosomes. Based on 200 nuclei from each of 30 normal specimens, the mean percentage of false-positive cells was 0.25 ± 0.39. Thirty-seven specimens from 10 patients were studied before treatment and two or more times at 4-month intervals after treatment with interferon-α2b (IFN-α2b) with or without ara-C. Based on 200 nuclei, the results of D-FISH in these specimens correlated closely with quantitative cytogenetics and accurately quantified disease within a few percent. We studied 6,000 nuclei for each of six specimens, three normal and three from patients with chronic myeloid leukemia (CML) in cytogenetic remission. The normal cutoff for 6,000 nuclei was 0.079% and patients in cytogenetic remission had residual disease ranging from 7 (0.117%) to 53 (0.883%) Ph-positive nuclei. We conclude that D-FISH can detect the Ph chromosome and its variant translocations and accurately quantify disease in CML at diagnosis and at all times after treatment, including cytogenetic remission.

scholarly journals Detection of chimeric BCR-ABL genes on bone marrow samples and blood smears in chronic myeloid and acute lymphoblastic leukemia by in situ hybridization

Blood ◽  
1994 ◽  
Vol 83 (7) ◽  
pp. 1922-1928 ◽  
Author(s):  
M Bentz ◽  
G Cabot ◽  
M Moos ◽  
MR Speicher ◽  
A Ganser ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Acute Lymphoblastic Leukemia ◽  
Yeast Artificial Chromosome ◽  
Lymphoblastic Leukemia ◽  
Philadelphia Chromosome ◽  
High Specificity ◽  
Breakpoint Cluster Region ◽  
Cluster Region ◽  
Blood Smears

The presence of BCR-ABL fusion genes has important diagnostic and prognostic implications in chronic myeloid leukemia (CML) and acute lymphoblastic leukemia (ALL). The CML-specific chimeric BCR-ABL gene with a break involving the major breakpoint cluster region (M-bcr) of the BCR-gene has been detected by means of fluorescence in situ hybridization (FISH). In this study, we present a FISH protocol that allows the detection of breaks in both the major and the minor breakpoint cluster region (m-bcr). Three hybridization signals of D107F9, a yeast artificial chromosome (YAC)-derived probe spanning the breakpoint regions of the BCR gene, were indicative of the translocation events. To increase the specificity further, this probe was combined with cos-abl 8, a cosmid probe flanking the breakpoint within the ABL gene for dual-color hybridization. Samples of 21 patients with CML, the ALL-derived cell line SUP-B15, and of seven patients with Philadelphia chromosome (Ph1)-positive ALL (three of them with breakpoints within m-bcr) were examined. BCR-ABL fusion was detected in all cases with high specificity (false-positive nuclei: mean, 0.1%). On cytogenetic preparations, the percentages of BCR-ABL- positive interphase cells ranged from 53% to 91%. Comparable efficiencies were achieved on blood smears. In conclusion, hybridization with D107F9 and cos-abl 8 allows unambiguous diagnosis of BCR-ABL genes and is likely to become an important tool for the monitoring of therapies in patients with CML and ALL.

scholarly journals Paternal origin of the rearranged major breakpoint cluster region in chronic myeloid leukemia

Blood ◽  
1994 ◽  
Vol 83 (12) ◽  
pp. 3445-3448 ◽  
Author(s):  
CE Litz ◽  
CM Copenhaver
Keyword(s):  
Chronic Myeloid Leukemia ◽  
Myeloid Leukemia ◽  
Restriction Site ◽  
Molecular Level ◽  
Philadelphia Chromosome ◽  
Chromosome 22 ◽  
Breakpoint Cluster Region ◽  
Cluster Region ◽  
Cytogenetic Studies ◽  
Restriction Site Polymorphisms

Abstract The Philadelphia chromosome, t(9;22), is present in virtually all cases of chronic myeloid leukemia (CML). It has previously been shown by cytogenetic studies that the rearranged chromosome 22 in patients with CML is exclusively maternal in origin. To address this issue at a molecular level, the major breakpoint cluster region (M-bcr) on chromosome 22 was examined using Southern blot assays and M-bcr Pvu II and Mae II restriction site polymorphisms in three CML patients. In all three cases, the rearranged allele was paternal in origin. These results indicate that the paternally derived M-bcr allele may also be involved in the M-bcr rearrangement.

scholarly journals Fluorescent in-situ hybridization (FISH) for BCR/ABL in chronic myeloid leukemia after bone marrow transplantation

2001 ◽  
Vol 119 (1) ◽  
pp. 16-18
Author(s):  
Maria de Lourdes Lopes Ferrari Chauffaille ◽  
José Salvador Rodrigues Oliveira ◽  
Maura Romeo ◽  
José Kerbauy
Keyword(s):  
Bone Marrow ◽  
In Situ Hybridization ◽  
Bone Marrow Transplantation ◽  
Chronic Myeloid Leukemia ◽  
Marrow Transplantation ◽  
Myeloid Leukemia ◽  
Fluorescent In Situ Hybridization ◽  
Residual Disease ◽  
Philadelphia Chromosome

CONTEXT: Identification of Philadelphia chromosome or BCR/ABL gene rearrangement in chronic myeloid leukemia is important at diagnosis as well as after treatment. OBJECTIVE: To compare the results of karyotyping using fluorescent in-situ hybridization (FISH) upon diagnosis and 1 year after bone marrow transplantation in 12 patients. TYPE OF STUDY: Diagnostic test and residual disease detection. SETTING: Hematology and Hemotherapy Department, Federal University of São Paulo/Escola Paulista de Medicina, São Paulo, Brazil. SAMPLE: 12 patients with chronic myeloid leukemia at diagnosis and 1 year after bone marrow transplantation. DIAGNOSTIC TEST: Karyotyping was done in the usual way and the BCR/ABL gene-specific probe was used for FISH. MAIN MEASUREMENTS: Disease at diagnosis and residual. RESULTS: At diagnosis, 10 patients presented t(9;22)(q34.1;q11) as well as positive FISH. Two cases did not have metaphases but FISH was positive. After bone marrow transplantation, 8 patients presented normal karyotype, 1 had persistence of identifiable Philadelphia chromosome and 3 had no metaphases. Two cases showed complete chimera and 2 had donor and host cells simultaneously. FISH was possible in all cases after bone marrow transplantation and confirmed the persistence of identifiable Philadelphia chromosome clone in one patient, and identified another that did not present metaphases for analysis. Cases that showed mixed chimera in karyotype were negative for BCR/ABL by FISH. CONCLUSION: The applicability of FISH is clear, particularly for residual disease detection. Classical and molecular cytogenetics are complementary methods.

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