FLUORESCENCE IIN SIITU HYBRIIDIIZATIION ANALYSIIS OF ADULT AML ACCORDIING TO RECENT UPDATES OF CLIINIICAL CYTOGENETIICS GUIIDELIINES AND QUALIITY ASSURANCE
Samar Magdy EL Sherif;
Abstract
SUMMARY AND CONCLUSION
C
ytogenetic analysis is one of the most powerful independent prognostic indicators in acute myeloid leukemia (AML), which serves to identify biologically distinct subsets of disease and has been widely adopted toprovide the framework for risk-adapted treatment approaches.
Between 10%-40% of newly diagnosed AML patients do not achieve complete remission with intensive induction therapy and are there for categorized as primary refractory or resistant. Few of these patients can be cured with conventional salvage therapy. They need to be evaluated regarding eligibility for allogeneic hematopoietic stem cell transplantation (HSCT) as this is currently the treatment with the highest probability of cure.
In the light of this, this work aimed to detect specific chromosomal mutations in acute myeloid leukemia patients, according to recent updates of professional guidelines of clinical cytogenetics, by using Fluorescence in situ Hybridization (FISH) and assess their relation with other prognostic factors and therapeutic response.
To achieve this aim, the present work was carried on 41 newly diagnosed adult AMLpatients. They were recruited from the Hematology oncology Unit, Ain Shams University hospitals and outpatients, during the period from June 2014 to May 2015. Out of the 41 patients, Sixteen (39%) were males and twenty five (61%) were females, with male to female ratio of (1: 1.6). Their age ranged from 19 to 71 years with amean (45±26) years. Diagnosis was based on standard morphologic, cytochemical and immunophenotyping criteria.
Informed consent was obtained from patients to use their samples in this study.Patients were evaluated at day 28 of therapy to assess therapeutic response.
All Patients were subjected to the following: Complete history taking and through clinical examination, Laboratory investigations including: Complete blood count using LH750 (Beckman coulter), examination of Leishman’s stained peripheral blood films, bone marrow aspiration and examination of Leishman’s stained bone marrow smears, cytochemical studies using myeloperoxidase stain. Immunophenotyping of bone marrow or peripheral blood samples using EPICS XL coulter flow cytometer to detect the FAB category.Fluorescence in situ hybridization analysis (FISH)cytogenetic analysis using the following probes:
Centromeric probe (CEP)8 for chromosome 8 to detect trisomy 8, LSI 5q31 with LSI 5p15.2 control for detection of deletion 5q and monosomy 5, LSI 7q31with centromeric control for detection of deletion 7q and monosomy7, LSI RUNX1-RUNX1T1 for t (8; 21) (q22; q22), LSI PML-RARA for t (15; 17) (q22; q12), RARA break apart probe for detection of 17q rearrangements, LSI BCR-ABL for t (9; 22) (q34; q11), LSI CBFB-MYH11forinv (16) (p13.1q22), LSI MLL break apart for 11q23 rearrangements andLSI EVI1break apart for inv (3q26).
Regarding the clinical presentation, out of the 41 patients, fourteen (34.1%) patients presented with hepatosplenomegaly and five (12.2%) patients presented with splenomegaly only. No CNS infiltration could be detected in all patient.
All the patients were anaemic with Hb level ranged from 4.3 to 9.6 g/dl with a mean value of (7±2.7) g/dl, the TLC ranged from 1.3 to 101 x109/L with a median value of (51.2±49.9) x109/L, the platelets count ranged from 30 to 101 x10^9/L with a mean value of (65.5±35.5) x109/L. The absolute PB blast ranged from 2 to 67 x10^9/L with a median value of (34.5±32.5) x109/L.According to FAB classification, one (2.4%) patients were classified as M0 subtype, Tweenty one (51.2%) patients were M1/M2 subtype, Six (14.6%) patients were M3, Seven (17.1%) were M4 and Six (14.6%) patients were M5.Bone marrow blasts ranged between 31 to 91% with a mean value of(61±30). Auer rods are frequently found and appear as a single long and sharp rod with tapered ends especially in AML M2.
Bone marrow examination showed distinct morphology for each subtype, as abnormal promyelocytespredominate both hypergranular or atypical microgranular (hypogranular) type with variable nuclear size and irregular contour in cases of AML-M3 &M3V. MonobIasts and promonocytes weretypically predominated in AML M4 and M5. Myelodysplastic changes were detected in the form of hypogranularity and hyposegmentation (pseudo-Pelger) in AML-M2 especially with t (8; 21), while multilineage dysplasia were seen in other cases especially with EVI (3q26) rearrangements. Moreover, the immunophenotypic analysis revealed one case (2.4 %) as M0, 21 cases (51.2 %) as M1/M2, 6 cases (14.6 %) as M3, 7 cases (17.1 %) as M4 and 6 cases (14.6 %) as M5.
C
ytogenetic analysis is one of the most powerful independent prognostic indicators in acute myeloid leukemia (AML), which serves to identify biologically distinct subsets of disease and has been widely adopted toprovide the framework for risk-adapted treatment approaches.
Between 10%-40% of newly diagnosed AML patients do not achieve complete remission with intensive induction therapy and are there for categorized as primary refractory or resistant. Few of these patients can be cured with conventional salvage therapy. They need to be evaluated regarding eligibility for allogeneic hematopoietic stem cell transplantation (HSCT) as this is currently the treatment with the highest probability of cure.
In the light of this, this work aimed to detect specific chromosomal mutations in acute myeloid leukemia patients, according to recent updates of professional guidelines of clinical cytogenetics, by using Fluorescence in situ Hybridization (FISH) and assess their relation with other prognostic factors and therapeutic response.
To achieve this aim, the present work was carried on 41 newly diagnosed adult AMLpatients. They were recruited from the Hematology oncology Unit, Ain Shams University hospitals and outpatients, during the period from June 2014 to May 2015. Out of the 41 patients, Sixteen (39%) were males and twenty five (61%) were females, with male to female ratio of (1: 1.6). Their age ranged from 19 to 71 years with amean (45±26) years. Diagnosis was based on standard morphologic, cytochemical and immunophenotyping criteria.
Informed consent was obtained from patients to use their samples in this study.Patients were evaluated at day 28 of therapy to assess therapeutic response.
All Patients were subjected to the following: Complete history taking and through clinical examination, Laboratory investigations including: Complete blood count using LH750 (Beckman coulter), examination of Leishman’s stained peripheral blood films, bone marrow aspiration and examination of Leishman’s stained bone marrow smears, cytochemical studies using myeloperoxidase stain. Immunophenotyping of bone marrow or peripheral blood samples using EPICS XL coulter flow cytometer to detect the FAB category.Fluorescence in situ hybridization analysis (FISH)cytogenetic analysis using the following probes:
Centromeric probe (CEP)8 for chromosome 8 to detect trisomy 8, LSI 5q31 with LSI 5p15.2 control for detection of deletion 5q and monosomy 5, LSI 7q31with centromeric control for detection of deletion 7q and monosomy7, LSI RUNX1-RUNX1T1 for t (8; 21) (q22; q22), LSI PML-RARA for t (15; 17) (q22; q12), RARA break apart probe for detection of 17q rearrangements, LSI BCR-ABL for t (9; 22) (q34; q11), LSI CBFB-MYH11forinv (16) (p13.1q22), LSI MLL break apart for 11q23 rearrangements andLSI EVI1break apart for inv (3q26).
Regarding the clinical presentation, out of the 41 patients, fourteen (34.1%) patients presented with hepatosplenomegaly and five (12.2%) patients presented with splenomegaly only. No CNS infiltration could be detected in all patient.
All the patients were anaemic with Hb level ranged from 4.3 to 9.6 g/dl with a mean value of (7±2.7) g/dl, the TLC ranged from 1.3 to 101 x109/L with a median value of (51.2±49.9) x109/L, the platelets count ranged from 30 to 101 x10^9/L with a mean value of (65.5±35.5) x109/L. The absolute PB blast ranged from 2 to 67 x10^9/L with a median value of (34.5±32.5) x109/L.According to FAB classification, one (2.4%) patients were classified as M0 subtype, Tweenty one (51.2%) patients were M1/M2 subtype, Six (14.6%) patients were M3, Seven (17.1%) were M4 and Six (14.6%) patients were M5.Bone marrow blasts ranged between 31 to 91% with a mean value of(61±30). Auer rods are frequently found and appear as a single long and sharp rod with tapered ends especially in AML M2.
Bone marrow examination showed distinct morphology for each subtype, as abnormal promyelocytespredominate both hypergranular or atypical microgranular (hypogranular) type with variable nuclear size and irregular contour in cases of AML-M3 &M3V. MonobIasts and promonocytes weretypically predominated in AML M4 and M5. Myelodysplastic changes were detected in the form of hypogranularity and hyposegmentation (pseudo-Pelger) in AML-M2 especially with t (8; 21), while multilineage dysplasia were seen in other cases especially with EVI (3q26) rearrangements. Moreover, the immunophenotypic analysis revealed one case (2.4 %) as M0, 21 cases (51.2 %) as M1/M2, 6 cases (14.6 %) as M3, 7 cases (17.1 %) as M4 and 6 cases (14.6 %) as M5.
Other data
| Title | FLUORESCENCE IIN SIITU HYBRIIDIIZATIION ANALYSIIS OF ADULT AML ACCORDIING TO RECENT UPDATES OF CLIINIICAL CYTOGENETIICS GUIIDELIINES AND QUALIITY ASSURANCE | Other Titles | تقنية التهجين الموضعى بالوميض الفللوري فى مرضى سرطان الدم النخاعى الحادفي البالغين طبقا إلى تحديثات المبادئ التوجيهيه للوراثة الخلوية وضمان الجودة | Authors | Samar Magdy EL Sherif | Issue Date | 2015 |
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