Autologous peripheral blood stem cell transplantation in acute myeloblastic leukaemia and myelodysplastic syndrome patients: evaluation of tumour cell contamination of leukaphereses by cytogenetic and molecular methods (original) (raw)

Successful peripheral blood stem cell transplantation for myelodysplastic syndrome

Bone Marrow Transplant, 1999

Keywords: autologous peripheral blood stem cell transplantation; myelodysplastic syndrome; WT1 gene expression The Wilms' tumor gene (WT1 gene), which acts as a tumor suppressor of Wilms' tumor and as a transcription factor for several growth and differentiation factors, is expressed in blast cells of acute leukemia and measurement of the WT1 level is useful for detecting MRD in acute leukemia. Recently, it has been demonstrated that WT1 gene expression is detectable in the blasts of MDS at diagnosis and that the level is significantly increased in patients with RAEB and RAEB-T. Myelodysplastic syndrome (MDS) represents clonal proliferation and maturation of hematopoietic stem cells, and is classified into five categories. Two of these, refractory anemia with excess of blasts (RAEB) and RAEB in transformation (RAEB-T), are associated with a high risk of developing acute leukemia. Such patients are usually treated with conventional chemotherapy but the results are unsatisfactory.

New clonal karyotypic abnormalities acquired following autologous bone marrow transplantation for acute myeloid leukemia do not appear to confer an adverse prognosis

Bone Marrow Transplantation, 1998

of myelodysplasia or secondary AML 5,6 and consequently a shortened survival. 7 Indeed, the development of cytogenetic abnormalities and MDS following ABMT for lymphoma We undertook a retrospective review of all 76 patients with AML transplanted between August 1986 and appear to be related to exposure to alkylating agents during conventional-dose chemotherapy rather than the high-March 1995 at our center. All patients received melphalan (140-160 mg/m 2), etoposide (60 mg/kg) and total dose regimen. 8 As induction and consolidation chemotherapy for AML body irradiation. All patients had bone marrow cytogenetic analysis at regular intervals following ABMT. do not generally include alkylating agents, we were interested in determining the incidence and significance of new The primary study end point was the development of the new cytogenetic abnormalities. Secondary end isolated cytogenetic abnormalities after ABMT for AML. points were the development of myelodysplasia (MDS) or AML. Sixty-two of 77 patients were alive at least 6 months post transplant. Cytogenetic abnormalities Patients and methods developed in 7/62 patients (11%) following ABMT. No patients demonstrated MDS or AML. At a median of We evaluated all patients undergoing ABMT for AML at our center between 1986 and 1995 to determine the fre-30 months after development of the cytogenetic abnormality, only one patient developed features suggestive quency and clinical significance of new clonal karyotypic abnormalities following transplantation. Patients Ͻ61 years but not diagnostic of MDS. All seven patients remain alive and leukemia-free up to 70 months after detection of age with ECOG performance status Ͻ2 were candidates for ABMT. Patients with persistent cytogenetic abnormali-of the abnormal clone. There was no increased incidence of cytogenetic abnormalities developing in patients ties or morphologic evidence of myelodysplasia following induction therapy were not eligible for ABMT. Intensive receiving a purged autograft. New cytogenetic abnormalities are frequent following ABMT for AML but do therapy and autotransplantation were performed in first or subsequent remission or early relapse with a marrow auto-not appear to predict development of myelodysplasia or acute myeloid leukemia. These abnormalities may relate graft collected in morphologic and cytogenetic remission. The intensive therapy regimen consisted of intravenous to use of total body radiation as part of the high-dose therapy. melphalan (140-160 mg/m 2), etoposide (VP-16) (60 mg/kg) and total body irradiation (TBI). The etoposide was Keywords: bone marrow transplantation; cytogenetics; acute myeloid leukemia; myelodysplasia administered as an infusion over 5 h for patients with good cardiac function and over 32 h for patients with left ventricular ejection fraction Ͻ45%. Patients in first complete remission were given a single fraction of TBI at a median Clonal cytogenetic abnormalities occur in the bone marrow rate of 50 cGy/min (range 30-60) to a total dose of 500 cGy of 50-90% of patients with acute myeloid leukemia (AML) while those in other disease states received six fractions of at the time of diagnosis and are strongly implicated in its 200 cGy administered at the same rate (total dose 1200 pathogenesis. 1,2 Furthermore, recurrence of the initial cGy). Patients were assessed at 3-monthly intervals for the malignant cytogenetic clone in the marrow of patients after first 2 years following ABMT and at 6-monthly intervals conventional-dose induction chemotherapy is usually assothereafter. Follow-up studies included a complete physical ciated with morphologic relapse. 3,4 examination, complete blood count and bone marrow aspir-New clonal cytogenetic abnormalities appearing after ation with cytogenetic analysis. Cytogenetic studies were conventional chemotherapy for lymphoma or other maligperformed according to standard methods with the karyonancies are generally felt to be associated with a high risk types described according to the international system for cytogenetic nomenclature (ISCN). 9 We defined new cytogenetic abnormalities following ABMT as persistent clonal

Donor Cell-derived Acute Myeloblastic Leukemia After Allogeneic Peripheral Blood Hematopoietic Stem Cell Transplantation for Juvenile Myelomonocytic Leukemia

Journal of Pediatric Hematology/Oncology, 2006

Despite its rarity, donor cell leukemia (DCL) is a most intriguing entity. We report here the case of a 5 year-old girl with juvenile myelomonocytic leukemia and normal female karyotype who developed acute myeloblastic leukemia with a karyotype of 46, X, t(X; 7) (p21; p11.2), der(7) t(3; 7) (q13.3; q22) 5 months after peripheral blood hematopoietic stem cell transplantation from her HLA-matched sister. We performed the analysis of short tandem repeat sequence markers to DNA obtained from donor peripheral blood, patient's peripheral blood including leukemic blasts and patient's hair root. This analysis showed that the leukemic blood DNA matched the donor blood DNA and not the patient's DNA, thus confirming DCL. To our knowledge, this is the first case of DCL after peripheral blood SCT for juvenile myelomonocytic leukemia.

Periodic morphologic, cytogenetic and clonality evaluation after autologous peripheral blood progenitor cell transplantation in patients with lymphoproliferative malignancies

Haematologica, 2002

Myelodysplastic syndrome (MDS), secondary acute myeloid leukemia (sAML) and clonal karyotypic abnormalities, have been recognized as relatively frequent and potentially serious complications of autologous peripheral blood progenitor cell transplantation (PBPCT) for Hodgkin's disease (HD), non-Hodgkin's lymphoma (NHL) or multiple myeloma (MM). We analyzed 66 patients, undergoing PBPCT for HD, NHL, MM or chronic lymphocytic leukemia (CLL). Patients reported in this study had to be in continuous complete remission after transplantation without receiving chemo-radiotherapy or other biological response modifiers, had to show absence of cytogenetic abnormalities and myelodysplastic features at transplantation and had to have at least 12 months of follow-up. We evaluated the bone marrow, peripheral blood, cytogenetics and clonality (HUMARA) 12 months after the transplant and thereafter every 12 months or every 6 months if lineage dysplasia, clonal or cytogenetic abnormalities were ...

Therapy-related myelodysplasia and/or acute myeloid leukaemia after autologous haematopoietic progenitor cell transplantation in a prospective single centre cohort of 221 patients

British Journal of Haematology, 2003

To evaluate the incidence and the predictive signs of therapy-related myelodysplasia and/or acute myeloid leukaemia (tMDS/tAML), we undertook a prospective study over a 4-year period of 221 patients who underwent autologous haematopoietic progenitor cell transplantation. Only seven patients (3AE1%) were identified to have tMDS/ tAML. Peripheral cytopenia was the first sign; diagnosis could be achieved by cytological analysis of bone marrow smears using World Health Organization criteria. All patients presented with bi-or trilineage dysplasia. Haematopoietic reconstitution was significantly delayed in patients progressing to tMDS/tAML compared with the control group. Typical cytogenetic abnormalities were observed in five of seven patients. The mean time interval between transplantation and cytological diagnosis, or detection of cytogenetic abnormalities, was 20AE0 months and 31AE2 months respectively. Pantelomeric fluorescence analysis using quantitative fluorescence in situ hybridization enabled us to make two major observations: (i) the fluorescence intensity in metaphases of all autografted patients was weak, and highly variable between tMDS patients; (ii) a drastic reduction of the telomere fluorescence intensity was observed in two patients who rapidly evolved to acute leukaemia. In conclusion, early detection of tMDS/tAML could be achieved by close follow-up of the bone marrow repopulation, and confirmed by cytological bone marrow examination and cytogenetic study. Our results address the implication of several factors, such as the initial telomeric status, and the effect of cytogenetic abnormalities and clonal expansion on bone marrow repopulation.

Donor-cell myelodysplastic syndrome developing 13 years after marrow grafting for aplastic anemia

Cancer Genetics and Cytogenetics, 2003

Donor-cell-derived hematopoietic malignancy is a rare event after bone marrow transplantation. Most cases in the literature occurred within the first year. We present a rare case of a female patient who had a bone marrow transplant for severe aplastic anemia (SAA) at the age of two and a half years from her human leukocyte antigen-identical brother. She developed a myelodysplastic syndrome (refractory cytopenia with multilineage dysplasia) 12 years later. Initially, the malignant clone was of recipient origin, but within several months, progression to a clinically more aggressive refractory anemia with excess blasts (RAEB) was accompanied by the outgrowth of a new clone of donor origin. In this report we provide evidence proving that the patient's final malignant clone arose in donor cells: cytogenetic analysis of the marrow showed a male karyotype and a t(3;21)(q26;q21) in all 62 metaphases analyzed. Interphase fluorescence in situ hybridization showed that all identifiable cells contained the Y chromosome. We conclude that donor-cell-derived hematopoietic malignancy after bone marrow transplantation can occur even after many years. We believe that the 13 years that elapsed between the transplant and the development of RAEB in our case represent the longest latency period in the literature.

Cytogenetic survey of 31 patients treated with bone marrow transplantation for acute nonlymphocytic and acute lymphoblastic leukemias

Cancer Genetics and Cytogenetics, 1991

The authors report on a sequential cytagenetic study carried aut on 31 patients with acute leukemia (20 with acute lymphnblastic leukemia and 11 with acute non-lymphoc:ytic leukemia) who underwent hone marrow transplantation (BMTL Engraftment was documented in all patients with sex-mismatched donors and with donor constitutianal aberrations. During the followup, ranging from 6 ta 110 months, clinical and hematologic relapse was observed in 17 patients (35.5%). Five of these cases showed a normal karyatype. 3 were tff undefined relapse origin, 2 were aneuploid karyotypes, and one was donor (maleJ metaphoses. Cytogenetic and immunologic data in the latter patient were suggestive of relapse in donor cells.

Evaluation of the utility of peripheral blood vs bone marrow in karyotype and fluorescence in situ hybridization for myelodysplastic syndrome diagnosis

Journal of Clinical Laboratory Analysis

To clear the role of peripheral blood as a substitution for bone marrow in myelodysplastic syndrome and to evaluate the concordance between peripheral blood and bone marrow using karyotype and fluorescence in situ hybridization (FISH) methods. Methods: We examined 35 bone marrow (BM) and peripheral blood (PB) samples from myelodysplastic syndrome (MDS) patient using karyotype and FISH. Karyotype method for BM and PB samples performed using the standard protocol with an exception for peripheral blood in which growth factor for cultivation was not used. FISH testing was performed using a panel of MDS-associated probes to detect 20q12, 20qter, 5q31, 5q33, 5p15 and chromosome 7 and 8 centromeres. Results: Our results showed karyotypes of BM and PB are concordant in 74% of cases, while about 53% of these concordances were achieved from cases with normal karyotypes. However, the results of BM FISH were completely concordant with PB FISH. Conclusion: Although peripheral blood karyotype is not trustworthy for MDS diagnosis, examining peripheral blood, using the FISH method, could be useful for clinical monitoring.

Influence of cytogenetic abnormalities on outcome after allogeneic bone marrow transplantation for acute myeloid leukemia in first complete remission

Biology of Blood and Marrow Transplantation, 2002

Cytogenetic abnormalities detected at diagnosis are recognized as important in predicting response to chemotherapy in acute myeloid leukemia (AML). However, there is controversy concerning the prognostic significance of karyotype for outcome after allogeneic bone marrow transplantation (allo-BMT) performed in first complete remission (CR1). This single-institution report describes allo-BMT for AML in CR1 and the effect of diagnostic cytogenetic findings on the results of that treatment. Between August 1981 and December 1999, 93 patients underwent related donor (n = 82) or unrelated donor (n = 11) BMT. Conditioning and GVHD prophylaxis were achieved predominantly with busulfan and cyclophosphamide and with cyclosporine and methotrexate, respectively. Seventynine (85%) of 93 patients had successful marrow karyotyping at diagnosis, and the patients were categorized into 3 prognostic groups based on the British Medical Research Council AML 10 trial classification: 15 patients (19%) were classified as having favorable risk [inv(16), t(8;21), t(15;17)]; 55 (70%) as having intermediate risk [no abnormality, +8, +21, +22, del(7q), del(9q), 11q23 rearrangement, and other numerical or structural abnormalities]; and 9 (11%) as having adverse risk [-5, del(5q), -7, 3q rearrangements, ≥5 abnormalities, t(6;9), t(9;22)]. The median follow-up was 93 months (range, 16-241 months). The overall survival (OS) rate, event-free survival (EFS) rate, relapse rate, and treatment-related mortality (TRM) were not statistically different between the groups. The 5-year actuarial EFS rates for favorable, intermediate, and adverse risk groups were 58% (95% confidence interval [CI], 29%-79%), 58% (95% CI, 43%-70%), and 67% (95% CI 28%-88%), respectively. Reclassification of patients into cytogenetic prognostic subgroups according to Southwest Oncology Group criteria did not change these results. In univariate analysis, the only variable found to have a prognostic influence on OS (P = .04) and TRM (P = .03) was the type of donor (unrelated donor was linked to a worse prognosis), which was confirmed in multivariate analysis. Our study suggests that presentation karyotype has less prognostic significance for outcome following allo-BMT than for outcome following conventional chemotherapy. In particular, AML patients with poor prognostic cytogenetic changes in CR1 who are unlikely to be cured with chemotherapy alone may benefit from allo-BMT.