minimal residual disease

Technical Aspects of Minimal Residual Disease Detection by Multicolor Flow Cytometry in Acute Myeloid Leukemia Patients

AUTOIMMUNE THROMBOCYTOPENIA , ,

IV Galtseva, YuO Davydova, NM Kapranov, KA Nikiforova, EN Parovichnikova

National Research Center for Hematology, 4 Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167

For correspondence: Yuliya Olegovna Davydova, MD, PhD, 4 Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167; Tel.: 8(495)612-62-21; e-mail: juliya89mur@yandex.ru

For citation: Galtseva IV, Davydova YuO, Kapranov NM, et al. Technical Aspects of Minimal Residual Disease Detection by Multicolor Flow Cytometry in Acute Myeloid Leukemia Patients. Clinical oncohematology. 2021;14(3):503–12. (In Russ).

DOI: 10.21320/2500-2139-2021-14-4-503-512

ABSTRACT

Detection and monitoring of minimal residual disease (MRD) are essential components of programmed therapy.They are crucial for the choice of treatment strategy and for prognostic purposes practically in all hematologic diseases. MRD is often detected by multicolor flow cytometry, the method with fairly high specificity and sensitivity. However, to identify MRD in acute myeloid leukemia patients is one of the most challenging tasks flow cytometry specialists are faced with. Cytometric data analysis requires the expert knowledge of immunophenotype of all maturing bone marrow cells. Besides, MRD analysis in acute myeloid leukemia has not been standardized while approaches suggested by different studies vary considerably. The present paper reports the experience of MRD analysis, demonstrates the gating strategy, immunophenotype description of normal non-tumor hematopoietic cells, and presents some examples of MRD assessment. Additionally, panels of monoclonal antibodies are provided, along with an evaluation of their advantages and disadvantages.

Keywords: minimal residual disease, acute myeloid leukemias, flow cytometry, gating, immunophenotyping.

Received: June 9, 2021

Accepted: September 5, 2021

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Статистика Plumx английский

REFERENCES

  1. Cheson BD, Bennett JM, Kopecky KJ, et al. Revised recommendations of the international working group for diagnosis, standardization of response criteria, treatment outcomes, and reporting standards for therapeutic trials in acute myeloid leukemia. J Clin Oncol. 2003;21(24):4642–9. doi: 10.1200/JCO.2003.04.036.
  2. Pui CH, Campana D. New definition of remission in childhood acute lymphoblastic leukemia. Leukemia. 2000;14(5):783–5. doi: 10.1038/sj.leu.2401780.
  3. Schuurhuis GJ, Heuser M, Freeman S, et al. Minimal/measurable residual disease in AML: a consensus document from the European LeukemiaNet MRD Working Party. Blood. 2018;131(12):1275–91. doi: 10.1182/blood-2017-09-801498.
  4. Гальцева И.В., Давыдова Ю.О., Паровичникова Е.Н. Определение минимальной измеримой остаточной болезни у взрослых больных острыми лейкозами. Гематология и трансфузиология. 2020;65(4):460–72. doi: 10.35754/0234-5730-2020-65-4-460-472.
    [Galtseva IV, Davydova YO, Parovichnikova EN. Detection of measurable residual disease in adults with acute leukaemia. Russian journal of hematology and transfusiology. 2020;65(4):460–72. doi: 10.35754/0234-5730-2020-65-4-460-472. (In Russ)]
  5. Shen Z, Gu X, Mao W, et al. Influence of pre-transplant minimal residual disease on prognosis after Allo-SCT for patients with acute lymphoblastic leukemia: Systematic review and meta-analysis. BMC Cancer. 2018;18(1):755. doi: 10.1186/s12885-018-4670-5.
  6. Leung W, Pui C-H, Coustan-Smith E, et al. Detectable minimal residual disease before hematopoietic cell transplantation is prognostic but does not preclude cure for children with very-high-risk leukemia. Blood. 2012;120(2):468–72. doi: 10.1182/blood-2012-02-409813.
  7. Norkin M, Katragadda L, Zou F, et al. Minimal residual disease by either flow cytometry or cytogenetics prior to an allogeneic hematopoietic stem cell transplant is associated with poor outcome in acute myeloid leukemia. Blood Cancer J. 2017;7(12):634. doi: 10.1038/s41408-017-0007-x.
  8. Anthias C, Dignan FL, Morilla R, et al. Pre-transplant MRD predicts outcome following reduced-intensity and myeloablative allogeneic hemopoietic SCT in AML. Bone Marrow Transplant. 2014;49(5):679–83. doi: 10.1038/bmt.2014.9.
  9. Buckley SA, Wood BL, Othus M, et al. Minimal residual disease prior to allogeneic hematopoietic cell transplantation in acute myeloid leukemia: a meta-analysis. Haematologica. 2017;102(5):865–73. doi: 10.3324/haematol.2016.159343.
  10. Wood BL. Principles of minimal residual disease detection for hematopoietic neoplasms by flow cytometry. Cytometry B Clin Cytom. 2016;90(1):47–53. doi: 10.1002/cyto.b.21239.
  11. Wood BL. Multicolor immunophenotyping: human immune system hematopoiesis. Methods Cell Biol. 2004;75:559–76. doi: 10.1016/s0091-679x(04)75023-2.
  12. Wood BL. Flow cytometric monitoring of residual disease in acute leukemia. In: Czader M, ed. Hematological Malignancies. Methods in Molecular Biology (Methods and Protocols). Vol. 999. Totowa: Humana Press; 2013. pp. 123–36. doi: 10.1007/978-1-62703-357-2_8.
  13. Лобанова Т.И., Гальцева И.В., Паровичникова Е.Н. Исследование минимальной остаточной болезни у пациентов с острыми миелоидными лейкозами методом многоцветной проточной цитофлуориметрии (обзор литературы). Онкогематология. 2018;13(1):83–102. doi: 10.17650/1818-8346-2018-13-1-83-102.
    [Lobanova TI, Galtseva IV, Parovichnikova EN. Minimal residual disease assesment in patients with acute myeloid leukemia by multicolour flow cytometry (literature review). Oncohematology. 2018;13(1):83–102. doi: 10.17650/1818-8346-2018-13-1-83-102. (In Russ)]
  14. Tien HF, Wang CH. CD7 positive hematopoietic progenitors and acute myeloid leukemia and other minimally differentiated leukemia. Leuk Lymphoma. 1998;31(1–2):93–8. doi: 10.3109/10428199809057588.
  15. Jorgensen JL, Chen SS. Monitoring of minimal residual disease in acute myeloid leukemia: methods and best applications. Clin Lymphoma Myeloma Leuk. 2011;11(Suppl 1):S49–53. doi: 10.1016/j.clml.2011.03.023.
  16. Jaso JM, Wang SA, Jorgensen JL, Lin P. Multi-color flow cytometric immunophenotyping for detection of minimal residual disease in AML: past, present and future. Bone Marrow Transplant. 2014;49(9):1129–38. doi: 10.1038/bmt.2014.99.
  17. Buldini B, Maurer-Granofszky M, Varotto E, Dworzak MN. Flow-cytometric monitoring of minimal residual disease in pediatric patients with acute myeloid leukemia: recent advances and future strategies. Front Pediatr. 2019;7:412. doi: 10.3389/fped.2019.00412.
  18. Wood BL. Acute myeloid leukemia minimal residual disease detection: the difference from normal approach. Curr Protoc Cytom. 2020;93(1):e73. doi: 10.1002/cpcy.73.
  19. Ostendorf BN, Flenner E, Florcken A, Westermann J. Phenotypic characterization of aberrant stem and progenitor cell populations in myelodysplastic syndromes. PLoS One. 2018;13(5):e0197823. doi: 10.1371/journal.pone.0197823.
  20. Goardon N, Marchi E, Atzberger A, et al. Coexistence of LMPP-like and GMP-like leukemia stem cells in acute myeloid leukemia. Cancer Cell. 2011;19(1):138–52. doi: 10.1016/j.ccr.2010.12.012.
  21. Shameli A, Dharmani-Khan P, Luider J, et al. Exploring blast composition in myelodysplastic syndromes and myelodysplastic/myeloproliferative neoplasms: CD45RA and CD371 improve diagnostic value of flow cytometry through assessment of myeloblast heterogeneity and stem cell aberrancy. Cytom Part B: Clin Cytom. 2020:1–16. doi: 10.1002/cyto.b.21983. Epub ahead of print.
  22. Bill M, van Kooten Niekerk BP, Woll SP, et al. Mapping the CLEC12A expression on myeloid progenitors in normal bone marrow; implications for understanding CLEC12A-related cancer stem cell biology. J Cell Mol Med. 2018;22(4):2311–8. doi: 10.1111/jcmm.13519.
  23. Eissens DN, Spanholtz J, van der Meer A, et al. Defining early human NK cell developmental stages in primary and secondary lymphoid tissues. PLoS One. 2012;7(2):e30930. doi: 10.1371/journal.pone.0030930.
  24. Stetler-Stevenson M, Paiva B, Stoolman L, et al. Consensus guidelines for myeloma minimal residual disease sample staining and data acquisition. Cytom Part B: Clin Cytom. 2016;90(1):26–30. doi: 10.1002/cyto.b.21249.
  25. Palmieri R, Piciocchi A, Arena V, et al. Clinical relevance of- limit of detection (LOD) – limit of quantification (LOQ) – based flow cytometry approach for measurable residual disease (MRD) assessment in acute myeloid leukemia (AML). Blood. 2020;136(Suppl 1):37–8. doi: 10.1182/blood-2020-139557.

Diagnosis of Acute Lymphoblastic Leukemia Originating From T-Lineage Precursors and Approaches to Minimal Residual Disease Monitoring

LYMPHOID TUMORS , , ,

OA Chernysheva, LYu Grivtsova, IN Serebryakova, NA Kupryshina, EN Sholokhova, MA Shervashidze, AD Palladina, BV Kurdyukov, AV Popa, NN Tupitsyn

NN Blokhin National Medical Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478

For correspondence: Ol’ga Alekseevna Chernysheva, 24 Kashirskoye sh., Moscow, Russian Federation, 115478; Tel.: +7(499)324-14-30; e-mail: beznos.olga@gmail.com

For citation: Chernysheva OA, Grivtsova LYu, Serebryakova IN, et al. Diagnosis of Acute Lymphoblastic Leukemia Originating From T-Lineage Precursors and Approaches to Minimal Residual Disease Monitoring. Clinical oncohematology. 2019;12(1):79–85.

DOI: 10.21320/2500-2139-2019-12-1-79-85

ABSTRACT

Background. Minimal residual disease (MRD) is an independent prognostic factor in acute lymphoblastic leukemia (ALL) in children. The immunological assessment of MRD cell count is based on aberrant immunophenotype of tumor lymphoblasts. However, in the case of ALL originating from T-lineage precursor cells (T-ALL) no clear aberrancy criteria have been defined, yet. Flow-cytometric MRD assessment in T-ALL can be based on characteristics of normal T-cell ontogenesis, i.e. the absence of normal T-lineage precursor cells (T-LP) in bone marrow.

Aim. To assess the feasibility of immunological method of flow cytometry for MRD detection based on T-LP immunophenotype on Days 15 and 33 of treatment of T-ALL children.

Materials & Methods. The analysis included the data on primary immunophenotype and MRD assessment on Dayы 15 and 33 of treatment of 31 T-ALL patients in the age of 2–17 years. In the majority of cases (61.3 %) the cortical/thymic immuno-subvariant of ALL was detected, in the rest of cases (38.7 %) it was the pre-T-cell one. Diagnosis was based on cumulative results of morphocytochemical and immunological bone marrow analyses. Assessing the MRD state the morphological and immunological analyses of bone marrow aspirate were carried out in parallel with one and the same tube. All patients enrolled in the trial were treated at Scientific Research Institute of Pediatric Oncology and Hematology of NN Blokhin National Medical Cancer Research Center according to the ALL IC-BFM 2009 protocol.

Results. Our study demonstrated that at all therapy stages MRD can be assessed by the unified immunological method based on detecting cyCD3+CD7+/++smCD3– (T-LP) immunophenotype cells. It is important to ensure that the correct clones of monoclonal antibodies are used for detecting CD3 cytoplasmic and membrane molecules (UCHT1 and SK7, respectively). Standard risk group included no patients. The majority of patients (76.2 %) treated according to ALL IC-BFM 2009 protocol were assigned to medium risk group on Day 15 of treatment. By Day 33 a quarter of them (25 %) was included into high risk group.

Conclusion. The capabilities of multicolor flow cytometry allow for the most complete characterization of primary immunophenotype of tumor T-cell lymphoblasts for further search of leukemia-associated immunophenotypes. Specific ontogenesis features of normal T-cells enable unification of immunological approaches to MRD assessment at all stages of T-ALL therapy.

Keywords: T-lineage acute lymphoblastic leukemia, multicolor flow cytometry, minimal residual disease, leukemia-associated immunophenotype.

Received: June 21, 2018

Accepted: December 18, 2018

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REFERENCES

  1. Clavell LA, Gelber RD, Cohen HJ. et al. A. Four-agent induction and intensive asparaginase therapy for treatment of childhood acute lymphoblastic leukemia. N Engl J Med. 1986;315(11):657–63. doi: 10.1056/nejm198609113151101.

  2. Crist WM, Shuster JJ, Falletta J, et al. Clinical features and outcome in childhood T-cell leukemia-lymphoma according to stage of thymocyte differentiation: a Pediatric Oncology Group Study. Blood. 1988;72(6):1891–7.

  3. Ludwig WD, Harbott J, Bartram CR, et al. Incidence and prognostic significance of immunophenotypic subgroups in childhood acute lymphoblastic leukemia: experience of the BFM study 86. Rec Res Cancer Res. 1993;131:269–82. doi: 10.1007/978-3-642-84895-7_24.

  4. Pui CH, Behm FG, Crist WM. Clinical and biologic relevance of immunologic marker studies in childhood acute lymphoblastic leukemia. Blood. 1993;82(2):343–62.

  5. Uckun FM, Sensel MG, Sun L, et al. Biology and treatment of childhood T-lineage acute lymphoblastic leukemia. Blood. 1998;91(3):735–46.

  6. Бойченко Э.Г., Попов А.М., Макарова Т.А. и др. Острый лимфобластный лейкоз из ранних предшественников Т-клеток. Вопросы гематологии/онкологии и иммунопатологии в педиатрии. 2015;14(1):38–45.

    [Boichenko EG, Popov AM, Makarova TA, et al. Early T-cell precursor acute lymphoblastic leukemia. Voprosy gematologii/onkologii i immunopatologii v pediatrii. 2015;14(1):38–45. (In Russ)]

  7. Goldberg JM. Silverman LB, Levy DE, et al. Childhood T-cell acute lymphoblastic leukemia: the Dana-Farber Cancer Institute acute lymphoblastic leukemia consortium experience. J Clin Oncol. 20031;21(19):3616–22. doi:1200/JCO.2003.10.116.

  8. Moricke A, Reiter A, Zimmermann M, et al. Risk-adjusted therapy of acute lymphoblastic leukemia can decrease treatment burden and improve survival: treatment results of 2169 unselected pediatric and adolescent patients enrolled in the trial ALL-BFM 95. Blood. 2008;111(9):4477–89. doi: 10.1182/blood-2007-09-112920.

  9. Pui C-H, Evans WE. Treatment of acute lymphoblastic leukemia. N Engl J Med. 2006;354(2):166–78. doi: 10.1056/NEJMra052603.

  10. Basso G, Veltroni M, Valsecchi MG, et al. Risk of relapse of childhood acute lymphoblastic leukaemia is predicted by flow cytometric measurement of residual disease on day 15 bone marrow. J Clin Oncol. 2009;27(31):5168–74. doi: 10.1200/JCO.2008.20.8934.

  11. Borowitz M, Devidas M, Hunger SP, et al. Clinical significance of minimal residual disease in childhood acute lymphoblastic leukemia and its relationship to other prognostic factors: a Children’s oncology group study. Blood. 2008;111(12):5477–85. doi: 10.1182/blood-2008-01-132837.

  12. Schrappe M, Valsecchi MG, Bartram CR, et al. Late MRD response determines relapse risk overall and in subsets of childhood T-cell ALL: results of the AIEOP-BFM-ALL 2000 study. Blood. 2011;118(8):2077–84. doi: 10.1182/blood-2011-03-338707.

  13. Fronkova E, Mejstrikova E, Avigad S, et al. Minimal residual disease (MRD) analysis in the non-MRD-based ALL IC-BFM 2002 Protocol for childhood ALL: is it possible to avoid MRD testing? Leukemia. 2008;22(5):989–97. doi: 10.1038/leu.2008.22.

  14. van Dongen JJM, Seriu T, Panzer-Grumayer ER, et al. Prognostic value of minimal residual disease in acute lymphoblastic leukaemia in childhood. Lancet. 1998;352(9142):1731–8. doi: 10.1016/s0140-6736(98)04058-6.

  15. Безнос О.А., Гривцова Л.Ю., Попа А.В. и др. Определение минимальной остаточной болезни при В-линейных острых лимфобластных лейкозах с использованием подходов EuroFlow. Клиническая онкогематология. 2017;10(2):158–68. doi: 21320/2500-2139-2017-10-2-158-168.

    [Beznos OA, Grivtsova LYu, Popa AV, et al. Evaluation of Minimal Residual Disease in B-Lineage Acute Lymphoblastic Leukemia Using EuroFlow Approaches. Clinical oncohematology. 2017;10(2):158–68. doi: 10.21320/2500-2139-2017-10-2-158-168. (In Russ)]

  16. Jaffe ES, Campo E, Harris NL, et al. Introduction and overview of the classification of lymphoid neoplasms. In: Swerdlow SH, Campo E, Harris NL, et al. (eds) WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. edited by. Lyon: IARC Press; рр. 189–98.

  17. Borowitz MJ, Chan JKC, Bene M-C, Arber DA. T-lymphoblastic leukemia/lymphoma. In: Swerdlow SH, Campo E, Harris NL, et al. (eds) WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. edited by. Lyon: IARC Press; рр. 209–12.

  18. Gelin, C. Aubrit F, Phalipon A, et al. The E2 antigen, a 32 kd glycoprotein involved in T-cell adhesion processes, is the MIC2 gene product. EMBO J. 1989;8(11):3253–9. doi: 10.1002/j.1460-2075.1989.tb08485.x.

  19. Dworzak MN, Fritsch G, Buchinger P, et al. Flow cytometric assessment of human MIC2 expression in bone marrow, thymus, and peripheral blood. Blood. 1994;83(2):415–25.

  20. Hamilton GA, Fellinger EJ, Schratter I, et al. Characterization of a human endocrine tissue and tumor-associated Ewing’s sarcoma antigen. Cancer Res. 1988;48(21):6127–31.

  21. Levy R, Dilley J, Fox RI, et al. A human thymus-leukemia antigen defined by hybridoma monoclonal antibodies. Proc Natl Acad Sci USA. 1979;76(12):6552–56. doi: 10.1073/pnas.76.12.6552.

  22. Bodger MP, Francis GE, Delia D, et al. A monoclonal antibody specific for immature human hemopoietic cells and T lineage cells. J Immunol. 1981;127(6):2269–74.

  23. Roshal M, Fromm JR, Winter S, et al. Immaturity associated antigens are lost during induction for T cell lymphoblastic leukemia: implications for minimal residual disease detection. Cytometry B: Clin Cytom. 2010;78B(3):139–46. doi: 10.1002/cyto.b.20511.

  24. Janossy G, Coustan-Smith E, Campana D. The reliability of cytoplasmic CD3 and CD22 antigen expression in the immunodiagnosis of acute leukemia: a study of 500 Leukemia. 1989;3(3):170–81.

Magnetic Resonance Imaging of Bone Marrow and its Results as a Criterion for Administration of Maintenance Therapy After Auto-HSCT in Multiple Myeloma Patients

BONE MARROW TRANSPLANTATION , , , ,

MV Solov’ev, LP Mendeleeva, GA Yatsyk, NS Lutsik, MV Firsova, EG Gemdzhian, VG Savchenko

National Research Center for Hematology, 4 Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167

For correspondence: Maksim Valer’evich Solov’ev, 4 Novyi Zykovskii pr-d, Moscow, Russian Federation, 125167; Tel.: +7(495)612-31-92; е-mail: maxsolovej@mail.ru

For citation: Solov’ev MV, Mendeleeva LP, Yatsyk GA, et al. Magnetic Resonance Imaging of Bone Marrow and its Results as a Criterion for Administration of Maintenance Therapy After Auto-HSCT in Multiple Myeloma Patients. Clinical oncohematology. 2018;11(4):360–7.

DOI: 10.21320/2500-2139-2018-11-4-360-367

ABSTRACT

Aim. To evaluate the efficacy of maintenance therapy in multiple myeloma (MM) patients after autologous hematopoietic stem cell transplantation (auto-HSCT) based on the results of MRI of bone marrow.

Materials & Methods. The study included 32 MM patients aged 36 to 66 years (median 57 years) with complete remission after a single auto-HSCT. MRI of spine and pelvic bones was performed to identify the nature of bone marrow lesions and to determine the volume of tumor tissue on the day 100 after auto-HSCT. As maintenance therapy after auto-HSCT 14 patients received daily 15 mg lenalidomide in the period from day 1 to day 21 of the 28-day treatment course within 1 year. Monitoring of 18 patients was conducted without maintenance therapy. Statistical analysis included the assessment of progression-free survival (PFS) and relapse risk relationship to clinical and laboratory parameters.

Results. Twenty patients had a positive MRI (tumor volume > 1 cm3). Zero variation of MR signal in bone marrow and detection of a < 1 cm3 tumor were regarded as a negative MRI, which was the case in 12 patients. After reaching the negative MRI the best rates of 2-year PFS were registered: 100 % with maintenance therapy and 84 % without maintenance therapy. In patients with tumor load on MR scans the 2-year PFS significantly (p = 0.03) varied and accounted for 80 % in patients who received maintenance therapy vs. 33 % in patients without maintenance therapy. Administration of maintenance therapy after detecting residual tumor on MR scans on day 100 after auto-HSCT has a positive effect on PFS rates. Multivariate analysis confirmed the residual tumor on MR scans of bone marrow to be the most important parameter PFS depends on.

Conclusion. A negative MRI after auto-HSCT is a favourable prognostic factor determining a long-lasting (> 2 years) MM free period, despite the lack of maintenance therapy.

Keywords: multiple myeloma, magnetic resonance imaging (MRI), autologous hematopoietic stem cell transplantation (auto-HSCT), maintenance therapy, minimal residual disease.

Received: May 11, 2018

Accepted: August 29, 2018

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REFERENCES

  1. Менделеева Л.П., Вотякова О.М., Покровская О.С. и др. Национальные клинические рекомендации по диагностике и лечению множественной миеломы. Гематология и трансфузиология. 2016;61(1, прил. 2):1–24. doi: 10.18821/0234-5730-2016-61-1(Прил.2). [Mendeleeva LP, Votyakova OM, Pokrovskaya OS, et al. National clinical guidelines on diagnosis and treatment of multiple myeloma. Gematologiya i transfuziologiya. 2016;61(1, Suppl. 2):1–24. doi: 10.18821/0234-5730-2016-61-1(Прил.2). (In Russ)]

  2. Kumar SK, Rajkumar SV, Dispenzieri A, et Improved survival in multiple myeloma and the impact of novel therapies. Blood. 2008;111(5):2516–20. doi: 10.1182/blood-2007-10-116129.

  3. Mendeleeva LP, Solovev MV, Alexeeva A, at al. Multiple Myeloma in Russia (First Results of the Registration Trial). Blood. 2017;130(Suppl 1):5408.

  4. Passweg JR, Baldomero H, Bader Р, et al. Is the use of unrelated donor transplantation leveling off in Europe? The 2016 European Society for Blood and Marrow Transplant activity survey report. Bone Marrow Transplant. 2018. doi: 10.1038/s41409-018-0153-1. [Epub ahead of print]

  5. Passweg JR, Baldomero H, Bader P, et al. Hematopoietic stem cell transplantation in Europe 2014: more than 40 000 transplants annually. Bone Marrow Transplant. 2016;51(6):786–92. doi: 10.1038/bmt.2016.20.

  6. Gay F, Oliva S, Petrucci MT, et al. Autologous transplant vs oral chemotherapy and lenalidomide in newly diagnosed young myeloma patients: a pooled analysis. Leukemia. 2017;31(8):1727–34. doi: 10.1038/leu.2016.381.

  7. Roussel M, Lauwers-Cances V, Robillard N, et al. Front-Line Transplantation Program With Lenalidomide, Bortezomib, and Dexamethasone Combination As Induction and Consolidation Followed by Lenalidomide Maintenance in Patients With Multiple Myeloma: A Phase II Study by the Intergroupe Francophone du Myelome. J Clin Oncol. 2014;32(25):2712–7. doi: 10.1200/JCO.2013.54.8164.

  8. Moreau P, San Miguel J, Sonneveld P, et al. Multiple myeloma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol. 2017;28(Suppl 4):iv52–61. doi: 10.1093/annonc/mdx096.

  9. Syed YY. Lenalidomide: A Review in Newly Diagnosed Multiple Myeloma as Maintenance Therapy After ASCT. Drugs. 2017;77(13):1473–80. doi: 10.1007/s40265-017-0795-0.

  10. Goldschmidt H, Lokhorst HM, Mai EK, et al. Bortezomib before and after high-dose therapy in myeloma: long-term results from the phase III HOVON-65/GMMG-HD4 trial. Leukemia. 2018;32(2):383–90. doi: 10.1038/leu.2017.211.

  11. Rosinol L, Oriol A, Teruel AI, et al. Bortezomib and thalidomide maintenance after stem cell transplantation for multiple myeloma: a PETHEMA/GEM trial. Leukemia. 2017;31(9):1922–7. doi: 10.1038/leu.2017.35.

  12. Mellqvist UH, Gimsing P, Hjertner O, et al. Bortezomib consolidation after autologous stem cell transplantation in multiple myeloma: a Nordic Myeloma Study Group randomized phase 3 trial. Blood. 2013;121(23):4647–54. doi: 10.1182/blood-2012-11-464503.

  13. Sonneveld P, Schmidt-Wolf IG, van der Holt B, et al. Bortezomib induction and maintenance treatment in patients with newly diagnosed multiple myeloma: results of the randomized phase III HOVON-65/GMMG-HD4 trial. J Clin Oncol. 2012;30(24):2946–55. doi: 10.1200/JCO.2011.39.6820.

  14. McCarthy PL, Owzar K, Hofmeister C, et al. Lenalidomide after stem-cell transplantation for multiple myeloma. N Engl J Med. 2012;366(19):1770–81. doi: 10.1056/NEJMoa1114083.

  15. Attal M, Lauwers-Cances V, Marit G, et al. Lenalidomide Maintenance after Stem-Cell Transplantation for Multiple Myeloma. N Engl J Med. 2012;366(19):1782–91. doi: 10.1056/NEJMoa1114138.

  16. Palumbo A, Cavallo F, Gay F, et al. Autologous transplantation and maintenance therapy in multiple myeloma. N Engl J Med. 2014;371(10):895–905. doi: 10.1056/NEJMoa1402888.

  17. Solovev MV, Mendeleeva LP, Pokrovskaya OS, et al. Maintenance Therapy after Autologous Haematopoietic Stem Cell Transplantation (auto-HSCT) in Multiple Myeloma Patients with and without Minimal Residual Disease (MRD). Blood. 2016;128(22):2260.

  18. Solovev MV, Mendeleeva LP, Pokrovskaya OS, et al. The Duration of MRD-Negative Status in Multiple Myeloma (MM) Patients after Auto-HSCT Is a Criterion for Prolonged Remission without Maintenance Therapy. Blood. 2017;130(Suppl 1):3294.

  19. Dutoit JC, Verstraete KL. Whole-body MRI, dynamic contrast-enhanced MRI, and diffusion-weighted imaging for the staging of multiple myeloma. Skelet Radiol. 2017;46(6):733–50. doi: 10.1007/s00256-017-2609-6.

  20. Latifoltojar A, Hall‐Craggs M, Rabin N, et al. Whole body magnetic resonance imaging in newly diagnosed multiple myeloma: early changes in lesional signal fat fraction predict disease response. Br J Haematol. 2017;176(2):222–33. doi: 10.1111/bjh.14401.

  21. Lasocki A, Gaillard F, Harrison SJ. Multiple myeloma of the spine. Neuroradiol J. 2017;30(3):259–68. doi: 10.1177/1971400917699426.

  22. Bray TJ, Singh S, Latifoltojar A, et al. Diagnostic utility of whole body Dixon MRI in multiple myeloma: A multi-reader study. PLoS One. 2017;12(7):e0180562. doi: 10.1371/journal.pone.0180562.

  23. Sabour S. Whole-body ultra-low dose computed tomography in comparison with spinal magnetic resonance imaging in the assessment of disease in multiple myeloma; Methodological issues on Diagnostic value. Br J Haematol. 2017. doi: 10.1111/bjh.14849. [Epub ahead of print]

  24. Chantry A, Kazmi M, Barrington S, et al. Guidelines for the use of imaging in the management of patients with myeloma. Br J Haematol. 2017;178(3):380–93. doi: 10.1111/bjh.14827.

  25. Moulopoulos LA, Gika D, Anagnostopoulos A, et al. Prognostic significance of magnetic resonance imaging of bone marrow in previously untreated patients with multiple myeloma. Ann Oncol. 2005;16(11):1824–8. doi: 10.1093/annonc/mdi362.

  26. Mai EK, Hielscher T, Kloth JK, et al. Association between magnetic resonance imaging patterns and baseline disease features in multiple myeloma: analyzing surrogates of tumour mass and biology. Eur Radiol. 2016;26(11):3939–48. doi: 10.1007/s00330-015-4195-0.

  27. Walker R, Barlogie B, Haessler J, et al. Magnetic resonance imaging in multiple myeloma: diagnostic and clinical implications. J Clin Oncol. 2007;25(9):1121–8. doi: 10.1200/JCO.2006.08.5803.

  28. Richardson PG, Holstein SA, Schlossman RL, et al. Lenalidomide in combination or alone as maintenance therapy following autologous stem cell transplant in patients with multiple myeloma: a review of options for and against. Expert Opin Pharmacother. 2017;18(18):1975–85. doi: 10.1080/14656566.2017.1409207.

  29. Pulte ED, Dmytrijuk A, Nie L, et al. FDA Approval Summary: Lenalidomide as Maintenance Therapy After Autologous Stem Cell Transplant in Newly Diagnosed Multiple Myeloma. Oncologist. 2018;23(6):734–9. doi: 10.1634/theoncologist.2017-0440.

  30. Sengsayadeth S, Malard F, Savani BN, et al. Posttransplant maintenance therapy in multiple myeloma: the changing landscape. Blood Cancer J. 2017;7(3):e545. doi: 10.1038/bcj.2017.23.

  31. McCarthy PL, Holstein SA, Petrucci MT, et al. Lenalidomide Maintenance After Autologous Stem-Cell Transplantation in Newly Diagnosed Multiple Myeloma: A Meta-Analysis. J Clin Oncol. 2017;35(29):3279–89. doi: 10.1200/JCO.2017.72.6679.

  32. Sonneveld P, Avet-Loiseau H, Lonial S, et al. Treatment of multiple myeloma with high-risk cytogenetics: a consensus of the International Myeloma Working Group. Blood. 2016;127(24):2955–62. doi: 10.1182/blood-2016-01-631200.

  33. Sivaraj D, Green MM, Li Z, et al. Outcomes of Maintenance Therapy with Bortezomib after Autologous Stem Cell Transplantation for Patients with Multiple Myeloma. Biol Blood Marrow Transplant. 2017;23(2):262–8. doi: 10.1016/j.bbmt.2016.11.010.

  34. Jackson GH, Davies FE, Pawlyn C, et al. Lenalidomide Is a Highly Effective Maintenance Therapy in Myeloma Patients of All Ages; Results of the Phase III Myeloma XI Study. Blood. 2016;128(22):1143.

  35. Neben K, Lokhorst HM, Jauch A, et al. Administration of bortezomib before and after autologous stem cell transplantation improves outcome in multiple myeloma patients with deletion 17p. Blood. 2012;119(4):940–8. doi: 10.1182/blood-2011-09-379164.

  36. Jackson GH, Davies FE, Pawlyn C, et al. Lenalidomide Maintenance Significantly Improves Outcomes Compared to Observation Irrespective of Cytogenetic Risk: Results of the Myeloma XI Trial. Blood. 2017;130(Suppl 1):436.

  37. Mellqvist UH, Gimsing P, Hjertner O, et al. Bortezomib consolidation after autologous stem cell transplantation in multiple myeloma: a Nordic Myeloma Study Group randomized phase 3 trial. Blood. 2013;121(23):4647–54. doi: 10.1182/blood-2012-11-464503.

  38. Phase III Studies Present Additional Evidence for REVLIMID® (lenalidomide) as Maintenance Therapy in Multiple Myeloma. Available from: http://ir.celgene.com/releasedetail.cfm?releaseid=1003026 (accessed 2.05.2018).

  39. Соловьев М.В., Менделеева Л.П., Покровская О.С. и др. Множественная миелома: поддерживающая терапия после трансплантации аутологичных гемопоэтических стволовых клеток в зависимости от минимальной остаточной болезни. Терапевтический архив. 2017;89(7):25–31. doi: 10.17116/terarkh201789725-31.

    [Solovyev MV, Mendeleeva LP, Pokrovskaya OS, et al. Multiple myeloma: Maintenance therapy after autologous hematopoietic stem cell transplantation, depending on minimal residual disease. Terapevticheskii arkhiv. 2017;89(7):25–31. doi: 10.17116/terarkh201789725-31. (In Russ)]

 

Minimal Residual Disease and IGHV-Genes Mutational Status as the Main Predictors of Response to Bendamustine-Rituximab Therapy in Previously Untreated Chronic Lymphocytic Leukemia

LYMPHOID TUMORS , , , , , ,

YuV Mirolyubova, EA Stadnik, VV Strugov, TO Andreeva, TS Nikulina, YuV Virts, PA Butylin, AG Rumyantsev, AYu Zaritskey

VA Almazov National Medical Research Center, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341

For correspondence: Yuliya Vladimirovna Mirolyubova, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341; e-mail: juli9702@yandex.ru

For citation: Mirolyubova YuV, Stadnik EA, Strugov VV, et al. Minimal Residual Disease and IGHV-Genes Mutational Status as the Main Predictors of Response to Bendamustine-Rituximab Therapy in Previously Untreated Chronic Lymphocytic Leukemia. Clinical oncohematology. 2018;11(2):167–74.

DOI: 10.21320/2500-2139-2018-11-2-167-174

ABSTRACT

Background. In patients with chronic lymphocytic leukemia (CLL) the eradication of minimal residual disease (MRD) is a prognostic factor of overall survival (OS) and progression-free survival (PFS). IGHV mutational status has also independent prognostic value.

Aim. To analyse the impact of mutational status and MRD eradication in CLL patients after first-line standard BR (bendamustine + rituximab) immunochemotherapy.

Materials & Methods. The prospective study included patients with immunophenotypically confirmed CLL who had not previously received anticancer therapy. All patients were treated by BR combination from 2012 to 2015. MRD level was determined in 109 patients after completing the 3rd and the 6th treatment courses. IGHV mutational status data were available for 98 patients. IGHV mutational status was evaluated in accordance with ERIC recommendations. MRD was assessed by standardized method of 4-color flow cytometry.

Results. MRD negativity was achieved in 37 (34 %) out of 109 patients. MRD eradication correlated with the best PFS (p = 0.04). IGHV mutational status had a statistically significant impact on PFS (p = 0.02). In patients with MRD-negative response and IGHV mutation no unfavorable events occurred during the period of monitoring. Conversely, PFS rates in MRD-negative patients having no IGHV mutation and in MRD-positive patients with mutation were significantly worse. MRD eradication resulted in statistically significant improvement of PFS rates after completing 3 treatment courses, compared with the cases with MRD persistence regardless of residual malignant clone level (p = 0.01).

Conclusion. BR therapy as first-line treatment statistically improved PFS in patients who achieved MRD-negative remission after completing the 3rd treatment course. PFS was significantly higher in MRD-negative patients with IGHV mutation after 6 treatment courses. MRD negativity resulting from 6 BR therapies in patients having no IGHV mutation was not accompanied by PFS improvement. It follows that by itself MRD negativity cannot be considered to be a universal prognostic factor.

Keywords: chronic lymphocytic leukemia, minimal residual disease, bendamustine, rituximab, BR, IGHV, mutational status.

Received: December 29, 2017

Accepted: February 27, 2018

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REFERENCES

  1. Swerdlow SH, Campo E, Harris NL, et al. (eds) WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th edition. Lyon: IARC Press; 2008.
  2. Swerdlow SH, Campo E, Pileri SA, et al. The 2016 revision of the World Health Organization classification of lymphoid neoplasms. 2016;127(20):2375–90. doi: 10.1182/blood-2016-01-643569.
  3. Клинические рекомендации по обследованию и лечению больных хроническим лимфолейкозом. Под ред. И.В. Поддубной, В.Г. Савченко [электронный документ]. Доступно по: https://blood.ru/documents/clinical%20guidelines/26.%20klinicheskie-rekomendacii-2014-xll.pdf. Ссылка активна на 12.2017.[Poddubnaya IV, Savchenko VG, eds. Clinical guidelines in examination and treatment of patients with chronic lymphocytic leukemia [Internet]. Available from: https://blood.ru/documents/clinical%20guidelines/26.%20klinicheskie-rekomendacii-2014-xll.pdf. (accessed 27.12.2017) (In Russ)]
  4. Keating MJ, O’Brien S, Albitar M, et al. Early results of a chemoimmunotherapy regimen of fludarabine, cyclophosphamide, and rituximab as initial therapy for chronic lymphocytic leukemia. J Clin Oncol. 2005;23(18):4079–88. doi: 1200/JCO.2005.12.051.
  5. Eichhorst BF, Busch R, Hopfinger G, et al. Fludarabine plus cyclophosphamide versus fludarabine alone in first-line therapy of younger patients with chronic lymphocytic leukemia. 2006;107(3):885–91. doi: 10.1182/blood-2005-06-2395.
  6. Стадник Е.А., Никитин Е.А., Бидерман Б.В. и др. Ретроспективное сравнение эффективности и токсичности режимов лечения FC и FCR у первичных больных В-клеточным хроническим лимфолейкозом. Онкогематология. 2008;1–2:39–46.[Stadnik EA, Nikitin EA, Biderman BV, et al. Comparison of efficacy and toxicity of FC and FCR regimens in the treatment of primary B-cell chronic lymphocytic leukemia: a retrospective study. Onkogematologiya. 2008;1–2:39–46. (In Russ)]
  7. Vuillier F, Claisse JF, Vandenvelde C, et al. Evaluation of residual disease in B-cell chronic lymphocytic leukemia patients in clinical and bone-marrow remission using CD5-CD19 markers and PCR study of gene rearrangements. Leuk Lymphoma. 1992;7(3):195–204. doi: 10.3109/10428199209053623.
  8. Lenormand B, Bizet M, Fruchart C, et al. Residual disease in B-cell chronic lymphocytic leukemia patients and prognostic value. Leukemia. 1994;8(6):1019–26.
  9. Cabezudo E, Matutes E, Ramrattan M, et al. Analysis of residual disease in chronic lymphocytic leukemia by flow cytometry. Leukemia. 1997;11(11):1909–14. doi: 10.1038/sj.leu.2400835.
  10. Rawstron AC, Villamor N, Ritgen M, et al. International standardized approach for flow cytometric residual disease monitoring in chronic lymphocytic leukaemia. Leukemia. 2007;21(5):956–64. doi: 10.1038/sj.leu.2404584.
  11. Rawstron AC, Bottcher S, Letestu R, et al. Improving efficiency and sensitivity: European Research Initiative in CLL (ERIC) update on the international harmonised approach for flow cytometric residual disease monitoring in CLL. 2013;27(1):142–9. doi: 10.1038/leu.2012.216.
  12. Rawstron AC, Fazi C, Agathangelidis A, et al. A complementary role of multiparameter flow cytometry and high-throughput sequencing for minimal residual disease detection in chronic lymphocytic leukemia: an European Research Initiative on CLL study. Leukemia. 2016;30(4):929–36. doi: 10.1038/leu.2015.313.
  13. Луговская С.А., Почтарь М.Е., Наумова Е.В. Диагностика минимальной остаточной болезни при В-клеточном хроническом лимфолейкозе методом многопараметрической лазерной проточной цитофлюориметрии. Клиническая лабораторная диагностика. 2010;9:20–20а.[Lugovskaya SA, Pochtar’ ME, Naumova EV. Diagnosis of minimal residual diseases in B-cell chronic lympholeukemia by multiparametric laser flow cytofluorometry. Klinicheskaya laboratornaya diagnostika. 2010;9:20–20а. (In Russ)]
  14. Stehlikova O, Chovancova J, Tichy B, et al. Detecting minimal residual disease in patients with chronic lymphocytic leukemia using 8-color flow cytometry protocol in routine hematological practice. Int J Lab Hematol. 2014;36(2):165–71. doi: 10.1111/ijlh.12149.
  15. Bottcher S, Stilgenbauer S, Busch R, et al. Standardized MRD flow and ASO IGH RQ-PCR for MRD quantification in CLL patients after rituximab-containing immunochemotherapy: a comparative analysis. Leukemia. 2009;23(11):2007–17. doi: 10.1038/leu.2009.140.
  16. Thompson PA, Wierda WG. Eliminating minimal residual disease as a therapeutic end point: working toward cure for patients with CLL. Blood. 2016;127(3):279–86. doi: 10.1182/blood-2015-08-634816.
  17. Voena C, Ladetto M, Astolfi M, et al. A novel nested-PCR strategy for the detection of rearranged immunoglobulin heavy-chain genes in B cell tumors. Leukemia. 1997;11(10):1793–8. doi: 10.1038/sj.leu.2400801.
  18. Logan AC, Zhang B, Narasimhan B, et al. Minimal residual disease quantification using consensus primers and high-throughput IGH sequencing predicts post-transplant relapse in chronic lymphocytic leukemia. Leukemia. 2013;27(8):1659–65. doi: 10.1038/leu.2013.52.
  19. Pfitzner T, Engert A, Wittor H, et al. A real-time PCR assay for the quantification of residual malignant cells in B cell chronic lymphatic leukemia. Leukemia. 2000;14(4):754–66. doi: 10.1038/sj.leu.2401706.
  20. Hallek M, Cheson BD, Catovsky D, et al. Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: A report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood. 2008;111(12):5446–56. doi: 10.1182/blood-2007-06-093906.
  21. Guideline on the use of minimal residue disease as an endpoint in chronic lymphocytic leukaemia studies [Internet]. Available from: http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2014/12/WC500179047.pdf. (accessed 27.12.2017).
  22. Bottcher S, Ritgen M, Fischer K, et al. Minimal residual disease quantification is an independent predictor of progression-free and overall survival in chronic lymphocytic leukemia: a multivariate analysis from the randomized GCLLSG CLL8 trial. J Clin Oncol. 2012;30(9):980–8. doi: 10.1200/JCO.2011.36.9348.
  23. Kovacs G, Robrecht S, Fink AM, et al. Minimal Residual Disease Assessment Improves Prediction of Outcome in Patients With Chronic Lymphocytic Leukemia (CLL) Who Achieve Partial Response: Comprehensive Analysis of Two Phase III Studies of the German CLL Study Group. J Clin Oncol. 2016;34(31):3758–65. doi: 10.1200/JCO.2016.67.1305.
  24. Damle RN, Wasil T, Fais F, et al. IgV gene mutation status and CD38 expression as novel prognostic indicators in chronic lymphocytic leukemia. Blood. 1999;94(6):1840–7.
  25. Thompson PA, Tam CS, O’Brien SM, et al. Fludarabine, cyclophosphamide, and rituximab treatment achieves long-term disease-free survival in IGHV-mutated chronic lymphocytic leukemia. Blood. 2016;127(3):303–9. doi: 10.1182/blood-2015-09-667675.
  26. Kwok M, Rawstron A, Varghese A, et al. Minimal residual disease is an independent predictor for 10-year survival in CLL. Blood. 2016;128(24):2770–3. doi: 10.1182/blood-2016-05-714162.
  27. Strati P, Keating MJ, O’Brien SM, et al. Eradication of bone marrow minimal residual disease may prompt early treatment discontinuation in CLL. Blood. 2014;123(24):3727–32. doi: 10.1182/blood-2013-11-538116.
  28. Стругов В.В., Стадник Е.А., Румянцев А.М. и др. Влияние мутационного статуса IGHV-генов и стереотипности строения BCR на эффективность режима BR в первой линии терапии хронического лимфолейкоза. Клиническая онкогематология. 2017;10(2):141–9. doi: 10.21320/2500-2139-2017-10-2-141-149.[Strugov VV, Stadnik EA, Rumyantsev AM, et al. Effect of IGHV Gene Mutation Status and BCR Structure Stereotypy on Effectiveness of BR Regimen in First-Line Therapy of Chronic Lymphocytic Leukemia. Clinical oncohematology. 2017;10(2):141–9. doi: 10.21320/2500-2139-2017-10-2-141-149. (In Russ)]
  29. Никитин Е.А., Стадник Е.А., Лорие Ю.Ю. и др. Прогностическое значение мутационного статуса генов вариабельного региона иммуноглобулинов у больных хроническим лимфолейкозом, получавших комбинированную терапию флударабином и циклофосфаном. Терапевтический архив. 2007;79(7):66–70.[Nikitin EA, Stadnik EA, Lorie YuYu, et al. Prognostic significance of IGHV mutational status in chronic lymphocytic leukemia patients after combination therapy with fludarabine and cyclophosphan. Terapevticheskii arkhiv. 2007;79(7):66–70. (In Russ)]
  30. Cross NCP, White HE, Colomer D, et al. Laboratory recommendations for scoring deep molecular responses following treatment for chronic myeloid leukemia. Leukemia. 2015;29(5):999–1003. doi: 1038/leu.2015.29.
  31. Chanan-Khan A, Cramer P, Demirkan F, et al. Ibrutinib combined with bendamustine and rituximab compared with placebo, bendamustine, and rituximab for previously treated chronic lymphocytic leukaemia or small lymphocytic lymphoma (HELIOS): a randomised, double-blind, phase 3 study. Lancet Oncol. 2016;17(2):200–11. doi: 10.1016/S1470-2045(15)00465-9.
  32. Seymour JF, Ma S, Brander DM, et al. Venetoclax plus rituximab in relapsed or refractory chronic lymphocytic leukaemia: a phase 1b study. Lancet Oncol. 2017;18(2):230–40. doi: 10.1016/s1470-2045(17)30012-8.
  33. Stilgenbauer S, Chyla B, Eichhorst B, et al. Venetoclax in relapsed/refractory chronic lymphocytic leukemia (CLL) with 17p deletion: outcome and minimal residual disease from the full population of the pivotal M13-982 trial. Eur Hematol Assoc. 2017: Abstract S771.
  34. Stilgenbauer S, Morschhauser F, Wendtner C-M, et al. Phase Ib study (GO28440) of venetoclax with bendamustine/rituximab or bendamustine/obinutuzumab in patients with relapsed/refractory or previously untreated chronic lymphocytic leukemia. Blood. 2016;128(22): Abstract 4393.

Ibrutinib in the Treatment of Relapsed Chronic Lymphocytic Leukemia

NOVEL DRUGS , , ,

EA Stadnik, NS Timofeeva, VV Strugov, AYu Zaritskey

VA Almazov National Medical Research Center, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341

For correspondence: Elena Aleksandrovna Stadnik, PhD, 2 Akkuratova str., Saint Petersburg, Russian Federation, 197341; Tel.: +7(921)575-54-55; e-mail: elena_stadnik@mail.ru

For citation: Stadnik EA, Timofeeva NS, Strugov VV, Zaritskey AYu. Ibrutinib in the Treatment of Relapsed Chronic Lymphocytic Leukemia. Clinical oncohematology. 2018;11(1):42–9.

DOI: 10.21320/2500-2139-2018-11-1-42-49

ABSTRACT

Aim. To estimate ibrutinib efficacy in the treatment of first early CLL relapses and in patients with ≥ 2 lines of preceding therapy. Analysis of treatment results in patients with del(17p) and monitoring of minimal residual disease (MRD) and ibrutinib safety profile.

Materials & Methods. The analysis included the results of ibrutinib treatment in 31 patients with CLL. Twenty eight patients were treated by bendamustine and fludarabine containing regimens. The median prior treatment lines were 2 (range 1–10). The indications for the treatment initiation were the first early relapse in 51 % of cases (n = 16) and a relapse after 2 and more lines of therapy in 49 % of cases (n = 15). Ibrutinib was administered in mono- (n = 15) and combined therapy (n = 14) as well as in the R-BAC scheme (n = 2). Using FISH analysis del(17p) was found in 9 patients (34 %).

Results. Within the median follow up of 18 months (range 7–42+) the overall survival (OS) rate was reported to be 87 %, and the progression-free survival (PFS) rate was 77 %. The maximum MRD after a year of ibrutinib treatment was observed in case of combination with immunochemotherapy (e.g., R-BAC). Within the period of 18 months OS rate was 100 %, in the patient group with first early relapses and 66 % in the group with a relapse after 2 and more therapy lines (p = 0.02). Within the same examination period PFS was significantly higher (94 %) in the patient group with first early relapses compared to the previously treated patients (60 %) (p = 0.034). The most common adverse events were grade 1–2 purpura (30 %), grade 1–2 diarrhea (10 %), atrial fibrillation paroxysms (10 %) and arterial hypertension (10 %). Severe infectious complications registered in 6 % (n = 3) patients were successfully solved in the course of combined antibacterial and antimycotic treatment.

Conclusion. Ibrutinib was shown to be effective drug for treatment of relapsed CLL. The OS and PFS values were more favourable in patients with first early relapses compared to the patients with relapses after ≥ 2 lines of therapy prior to ibrutinib treatment. The maximum elimination of the tumor clone was observed after combined ibrutinib/immunochemotherapy treatment. The tolerance of ibrutinib was reported to be satisfactory with acceptable toxicity profile. No mortality due to infection complications was observed.

Keywords: chronic lymphocytic leukemia, first early relapse, del(17p), ibrutinib, minimal residual disease.

Received: August 20, 2017

Accepted: November 16, 2017

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REFERENCES

  1. Fornecker LM, Aurran-Schleinitz T, Michallet AS, et al. Salvage outcomes in patients with first relapse after fludarabine, cyclophosphamide, and rituximab for chronic lymphocytic leukemia: the French intergroup experience. Am J Hematol. 2015;90(6):511–4. doi: 10.1002/ajh.23999.
  2. Стадник Е.А., Стругов В.В., Андреева Т.О. и др. Эффективность комбинации бендамустина ритуксимаба в первой линии терапии ХЛЛ: результаты исследования BEN-001. Терапевтический архив. 2017;89(7):57–64. doi: 10.17116/terarkh201789757-64. [Stadnik EA, Strugov VV, Andreeva TO, et al. Efficacy of a bendamustine and rituximab combination in first-line therapy for chronic lymphocytic leukemia: Results of the BEN-001 study. Terapevticheskii arkhiv. 2017;89(7):57–64. doi: 10.17116/terarkh201789757-64. (In Russ)]
  3. Стругов В.В., Стадник Е.А., Вирц Ю.В. и др. Значение возраста и сопутствующих заболеваний в терапии хронического лимфолейкоза. Клиническая онкогематология. 2016;9(2):162–75. doi: 10.21320/2500-2139-2016-9-2-162-175. [Strugov VV, Stadnik EA, Virts YuV, et al. Role of Patient’s Age and Comorbidities in Therapy of Chronic Lymphocytic Leukemia. Clinical oncohematology. 2016;9(2):162–75. doi: 10.21320/2500-2139-2016-9-2-162-175. (In Russ)]
  4. Никитин Е.А., Стадник Е.А., Лорие Ю.Ю. и др. Прогностическое значение мутационного статуса генов вариабельного региона иммуноглобулинов у больных хроническим лимфолейкозом, получавших комбинированную терапию флударабином и циклофосфаном. Терапевтический архив. 2007;79(7):66–70. [Nikitin EA., Stadnik EA, Lorie YuYu, et al. Prognostic significance of immunoglobulin variable region mutations in B-CLL patients treated with combination therapy fludarabine plus cyclophosphamide. Terapevticheskii arkhiv. 2007;79(7):66–70. (In Russ)]
  5. Thompson PA, Tam CS, O’Brien SM, et al. Fludarabine, cyclophosphamide, and rituximab treatment achieves long-term disease-free survival in IGHV-mutated chronic lymphocytic leukemia. Blood. 2016;127(3):303–9. doi: 10.1182/blood-2015-09-667675.
  6. Fischer K, Bahlo J, Fink AM, et al. Long-term remissions after FCR chemoimmunotherapy in previously untreated patients with CLL: updated results of the CLL8 trial. Blood. 2016;127(2):208–15. doi: 10.1182/blood-2015-06-651125.
  7. Robbe P, Ridout K, Becq J, et al. Identifying High-Risk CLL to Predict Early Relapse after FCR Based Treatment Using Whole Genome Sequencing: First Results from the Genomics England CLL Pilot. Blood. 2016;128:2022.
  8. Castro JE, James DF, Sandoval-Sus JD, et al. Rituximab in combination with high-dose methylprednisolone for the treatment of chronic lymphocytic leukemia. Leukemia. 2009;23(10):1779–89. doi: 10.1038/leu.2009.133.
  9. Cramer P, Fink A-M, Busch R, et al. Second-line therapies of patients initially treated with fludarabine and cyclophosphamide or fludarabine, cyclophosphamide and rituximab for chronic lymphocytic leukemia within the CLL8 protocol of the German CLL Study Group. Leuk Lymphoma. 2013;54(8):1821–2. doi: 10.3109/10428194.2013.796050.
  10. Brown JR. The Treatment of Relapsed Refractory Chronic Lymphocytic Leukemia. Hematology. 2011;2011(1):110–8. doi: 10.1182/asheducation-2011.1.110.
  11. Tam CS, O’Brien S, Plunkett W. Long-term results of first salvage treatment in CLL patients treated initially with FCR (fludarabine, cyclophosphamide, rituximab). Blood. 2014;124(20):3059–64. doi: 10.1182/blood-2014-06-583765.
  12. Zenz T, Busch R, Fink A, et al. Genetics of patients with F-refractory CLL or early relapse after FC or FCR: Results from the CLL8 trial of the GCLLSG [Abstract]. Blood (ASH Annual Meeting Abstracts) 2010;116(21):2427.
  13. Stilgenbauer S, Zenz T, Winkler D, et al. Genomic Aberrations, VH Mutation Status and Outcome after Fludarabine and Cyclophosphamide (FC) or FC Plus Rituximab (FCR) in the CLL8 Trial. Blood. 2008;112(11): Abstract 781.
  14. Montserrat E, Dreger P. Treatment of Chronic Lymphocytic Leukemia With del(17p)/TP53 Mutation: Allogeneic Hematopoietic Stem Cell Transplantation or BCR-Signaling Inhibitors? Clin Lymph Myel Leuk. 2016;16:S74–S81. doi: 10.1016/j.clml.2016.02.013.
  15. Gladstone DE, Blackford A, Cho E, et al. The Importance of IGHV Mutational Status in del(11q) and del(17p) Chronic Lymphocytic Leukemia. Clin Lymph Myel Leuk. 2012;12(2):132–7. doi: 10.1016/j.clml.2011.12.005.
  16. The International CLL-IPI working group. An international prognostic index for patients with chronic lymphocytic leukaemia (CLL-IPI): a meta-analysis of individual patient data. Lancet Oncol. 2016;17(6):779–90. doi: 10.1016/S1470-2045(16)30029-8.
  17. Byrd JC, Brown JR, O’Brien S, et al. Ibrutinib versus ofatumumab in previously treated chronic lymphoid leukemia. N Engl J Med. 2014;371(3):213–23. doi: 10.1056/nejmoa1400376.
  18. O’Brien S, Jones JA, Coutre SE, et al. Ibrutinib for patients with relapsed or refractory chronic lymphocytic leukaemia with 17p deletion (RESONATE-17): a phase 2, open-label, multicentre study. Lancet Oncol. 2016;17(10):1409–18. doi: 10.1016/s1470-2045(16)30212-1.
  19. Visco C, Finotto S, Pomponi F, et al. The combination of rituximab, bendamustine, and cytarabine for heavily pretreated relapsed/refractory cytogenetically high-risk patients with chronic lymphocytic leukemia. Am J Hematol. 2013;88(4):289–93. doi: 10.1002/ajh.23391.
  20. Castegnaro S, Visco C, Chieregato K. et al. Cytosine arabinoside potentiates the apoptotic effect of bendamustine on several B- and T-cell leukemia/lymphoma cells and cell lines. Leuk Lymphoma. 2012;53(11):2262–8. doi: 10.3109/10428194.2012.688200.
  21. Brown JR, Hillmen P, O’Brien S, et al. Updated Efficacy Including Genetic and Clinical Subgroup Analysis and Overall Safety in the Phase 3 RESONATE TM Trial of Ibrutinib Versus Ofatumumab. Blood. 2014;124(21): Abstract 3331.
  22. Jones J, Coutr S, Byrd JC, et al. Evaluation of 243 patients with deletion 17p chronic lymphocytic leukemia treated with ibrutinib: a cross-study analysis of treatment outcomes. EHA Learning Center. 2016;135185: Abstract S429.
  23. Burger JA, Keating MJ, Wierda WG. Safety and activity of ibrutinib plus rituximab for patients with high-risk chronic lymphocytic leukemia: a single-arm, phase 2 study. Lancet Oncol. 2014;15(10):1090–9. doi: 10.1016/S1470-2045(14)70335-3.
  24. Fraser G, Cramer P, Hallek M, et al. Three-Year Follow-up of Patients With Previously Treated Chronic Lymphocytic Lymphoma (CLL) Receiving Ibrutinib Plus Bendamustine and Rituximab (BR) Versus Placebo Plus BR. An Update of the HELIOS Study. XVII iwCLL. 2017: Abstract 400.
  25. Burger JA, Tedeschi A, Bar PM. Ibrutinib as Initial Therapy for Patients with Chronic Lymphocytic Leukemia. N Engl J Med. 2015;373(25):2425–37. doi: 10.1056/NEJMoa1509388.

Evaluation of Minimal Residual Disease in B-Lineage Acute Lymphoblastic Leukemia Using EuroFlow Approaches

LYMPHOID TUMORS , ,

OA Beznos, LYu Grivtsova, AV Popa, MA Shervashidze, IN Serebryakova, OYu Baranova, EA Osmanov, NN Tupitsyn

NN Blokhin Russian Cancer Research Center, 24 Kashirskoye sh., Moscow, Russian Federation, 115478

For correspondence: Ol’ga Alekseevna Beznos, junior researcher, 24 Kashirskoye sh., Moscow, Russian Federation, 115478; Tel: 8(916)480-03-35; e-mail: beznos.olga@gmail.com

For citation: Beznos OA, Grivtsova LYu, Popa AV, et al. Evaluation of Minimal Residual Disease in B-Lineage Acute Lymphoblastic Leukemia Using EuroFlow Approaches. Clinical oncohematology. 2017;10(2):158–68 (In Russ).

DOI: 10.21320/2500-2139-2017-10-2-158-168

ABSTRACT

Background & Aims. Evaluation of the minimal residual disease (MRD) at different stages of chemotherapy is one of key prognostic factors and a factor of stratification of patients into risk groups in acute lymphoblastic leukemia (ALL). The MRD detection on Day 15 and at later stages is based on identifying blast cells with a leukemia-associated immune phenotype. The aim is to assess the potential of 8-color standardized EuroFlow panels and to detect individual criteria for MRD monitoring during primary diagnosis.

Materials & Methods. The analysis included data on the primary immune phenotype and MRD assessment during chemotherapy in 10 adults and 35 children with a confirmed diagnosis of B-cell precursors ALL.

Results. The ALL phenotype characteristics at the stage of primary diagnosis permit to make the most complete description of the of 8-color standardized EuroFlow panels. This gives an opportunity to select the most informative antigen combinations for further MRD monitoring. Combinations with CD58/CD38, CD81/СD9 antigen expression, as well as assessment of pan-myeloid CD13, CD33 antigen co-expression may be recommended as the most frequent aberrant immune phenotypes of blast cells in ALL. As for B-lineage progenitor cells in children on Day 15 of the induction therapy, a detection of TdT+ сyCD22+ cell population is necessary in addition to the quantification of CD10+ and/or CD34+ В-lineage progenitor cells.

Conclusion. Therefore, the 8-color standardized EuroFlow panels permit not only to characterize the primary ALL immune phenotype in details, but may also be widely used for MRD evaluation at all stages of chemotherapy.

Keywords: B-lineage acute lymphoblastic leukemia, multicolor flow cytometry, minimal residual disease.

Received: January 14, 2017

Accepted: January 29, 2017

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REFERENCES

  1. Borowitz MJ, Devidas M, Hunger SP, et al. Clinical significance of minimal residual disease in children acute lymphoblastic leukemia and its relationship to the prognostic factors: a Children’s Oncology Group study. Blood. 2008;111(12):5477–85. doi: 10.1182/blood-2008-01-132837.
  2. Dworzak MN, Froschl G, Printz D, et al. Prognostic significance and modalities of flow cytometric minimal residual disease detection in childhood acute lymphoblastic leukemia. Blood. 2002;99(6):1952–8. doi: 10.1182/blood.V99.6.1952.
  3. Basso G, Veltroni M, Valsecchi MG, et al. Risk of relapse of childhood acute lymphoblastic leukemia is predicted by flow cytometric measurement of residual disease on day 15 bone marrow. J Clin Oncol. 2009;27(31):5168–74. doi: 10.1200/jco.2008.20.8934.
  4. Coustan-Smith E, Sancho J, Behm FG, et al. Prognostic importance of measuring early clearance of leukemic cells by flow cytometry in childhood acute lymphoblastic leukemia. Blood. 2002;100(1);52–8. doi: 10.1182/blood-2002-01-0006.
  5. Coustan-Smith E, Ribeiro RC, Stow P, et al. A simplified flow cytometric assay identifies children with acute lymphoblastic leukemia who have a superior clinical outcome. Blood. 2006;108(1):97–102. doi: 10.1182/blood-2006-01-0066.
  6. Schrappe M, Reiter A, Ludwig WD, et al. Improved outcome in childhood acute lymphoblastic leukemia despite reduced use of antracyclines and cranial radiotherapy: results of trial ALL-BFM 90. Blood. 2000;95(11):3310–22.
  7. Гривцова Л.Ю., Попа А.В., Купрышина Н.А. и др. Оценка минимальной резидуальной болезни при острых лимфобластных лейкозах из В-линейных предшественников у детей методом трехцветной проточной цитометрии. Иммунология гемопоэза. 2008;5(2):8–33.
    [Grivtsova LYu, Popa AV, Kupryshina NA, et al. Detection of minimal residual disease in children with B-cell precursor acute lymphoblastic leukemia with simplified protocols. Immunologiya gemopoeza. 2008;5(2):8–33. (In Russ)]
  8. Гривцова Л.Ю., Попа А.В., Серебрякова И.Н., Тупицын Н.Н. К дальнейшей стандартизации определения остаточных бластных клеток в костном мозге детей с В-линейными острыми лимфобластными лейкозами на 15-й день индукционной терапии. Иммунология гемопоэза. 2011;8(1):35–54.
    [Grivtsova LYu, Popa AV, Serebryakova IN, Tupitsyn NN. To further standardization in detection of residual blasts in bone marrow of children with B-cell acute lymphoblastic leukemia on Day 15 of induction therapy. Immunologiya gemopoeza. 2011;8(1):35–54. (In Russ)]
  9. van Dongen JJM, van der Velden VHJ, Bruggemann M, et al. Minimal residual disease diagnostics in acute lymphoblastic leukemia: need for sensitive, fast, and standardized technologies. Blood. 2015;125(26):3996–4009. doi: 10.1182/blood-2015-03-580027.
  10. van Dongen JJM, Lhermitte L, Bottcher S, et al. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia. 2012;26(9):1908–75. doi: 10.1038/leu.2012.120.
  11. Локен М.Р., Уэлс Д.А. Определение клеток предшественников. Иммунология гемопоэза. 2010;7(1):8–22.
    [Loken MR, Wells DA. Enumeration of progenitor cells. Immunologiya gemopoeza. 2010;7(1):8–22 (In Russ)]
  12. Гривцова Л.Ю., Тупицын Н.Н. Иммунологическая оценка гемодилюции костного мозга при лабораторных исследованиях (на основании теста М. Локен). Медицинский алфавит. 2015;4(18):67–70.
    [Grivtsova LYu, Tupitsyn NN. Immunological evaluation of bone marrow hemodilution in laboratory test (based on M. Loken’s test). Meditsinskii alfavit. 2015;4(18):67–70. (In Russ)]
  13. Тупицын Н.Н., Гривцова Л.Ю., Купрышина Н.А. Иммунодиагностика опухолей крови на основании многоцветных (8-цветных панелей) Европейского консорциума по проточной цитометрии (Euroflow). Иммунология гемопоэза. 2015;13(1):31–62.
    [Tupitsyn NN, Grivtsova LYu, Kupryshina NA. Haematopoietic malignancies immune diagnostics based on Euroflow Consortium proposals: 8-color flow cytometry. Immunologiya gemopoeza. 2015;13(1):31–62. (In Russ)]
  14. Veltroni M, de Zen L, Sanzari MC, et al.; I-BFM-ALL-FCM-MRD-Study Group. Expression of CD58 in normal, regenerating and leukemic bone marrow B cells: implications for the detection of minimal residual disease in acute lymphocytic leukemia. J Hematol. 2003;88(11):1245–52.
  15. Romero-Ramırez H, Morales-Guadarrama MT, Pelayo R, et al. CD38 expression in early B-cell precursors contributes to extracellular signal-regulated kinase-mediated apoptosis. Immunology. 2014;144(2):271–81. doi: 10.1111/imm.12370.
  16. Tajima F, Deguchi T, Laver JH, et al. Reciprocal expression of CD38 and CD34 by adult murine hematopoietic stem cells. Blood. 2001;97(9):2618–24. doi: 10.1182/blood.V97.9.2618.
  17. Higuchi Y, Zeng H, Ogawa M. CD38 expression by hematopoietic stem cells of newborn and juvenile mice. Leukemia. 2003;17(1):171–4. doi: 10.1038/sj.leu.2402785.
  18. Carsetti R, Rosado MM, Wardmann H. Peripheral development of B cells in mouse and man. Immunol Rev. 2004;197(1):179–91. doi: 10.1111/j.0105-2896.2004.0109.x.
  19. Lamkin T, Brooks J, Annett G, et al. Immunophenotypic differences between putative hematopoietic stem cells and childhood B cell precursor acute lymphoblastic leukemia cells. Leukemia. 1994;8(11):1871–8.
  20. Chen JS, Coustan-Smith E, Suzuki T, et al. Identification of novel markers for monitoring minimal residual disease in acute lymphoblastic leukemia. Blood. 2001;97(7):2115–20. doi: 10.1182/blood.V97.7.2115.
  21. De Waele M, Renmans W, Jochmans K, et al. Different expression of adhesion molecules on CD34+ cells in AML and B-lineage ALL and their normal bone marrow counterparts. Eur J Haematol. 1999;63(3):192–201. doi: 10.1111/j.1600-0609.1999.tb01767.x.
  22. Dworzak MN, Fritsch G, Froschl G, et al. Four-Color Flow Cytometric Investigation of Terminal Deoxynucleotidyl Transferase–Positive Lymphoid Precursors in Pediatric Bone Marrow: CD79a Expression Precedes CD19 in Early B-Cell Ontogeny. Blood. 1998;92(9):3203–9.
  23. Coustan-Smith E, Song G, Clark C, et al. New markers for minimal residual disease detection in acute lymphoblastic leukemia. Blood. 2011;117(23):6267–76. doi: 10.1182/blood-2010-12-324004.
  24. Barrena S, Almeida J, Yunta M, et al. Aberrant expression of tetraspanin molecules in B-cell chronic lymphoproliferative disorders and its correlation with normal B-cell maturation. Leukemia. 2005;19(8):1376–83. doi: 10.1038/sj.leu.2403822.
  25. Тупицын Н.Н., Гривцова Л.Ю., Купрышина Н.А. Проточная цитометрия в онкогематологии. Часть I. Основы и нововведения в диагностике острых лейкозов. Клиническая онкогематология. 2012;5(1):42–7.
    [Tupitsyn NN, Grivtsova LYu, Kupryshina NA. Flow cytometry in hematology malignancies. Part I. ABC and news in acute leukemia diagnostics. Klinicheskaya onkogematologiya. 2012;5(1):42–7. (In Russ)]
  26. Shoham T, Rajapaksa R, Boucheix C, et al. The Tetraspanin CD81 Regulates the Expression of CD19 During B Cell Development in a Postendoplasmic Reticulum Compartment. J Immunol. 2003;171(8):4062–72. doi: 10.4049/jimmunol.171.8.4062.

Chronic Lymphocytic Leukemia: Prognostic Significance of Minimal Residual Disease and Potential of Modern Methods of Its Diagnosis and Therapy (Literature Review)

LYMPHOID TUMORS , ,

AYu Kuvshinov, SV Voloshin, IS Martynkevich, EV Kleina, MA Mikhaleva, KM Abdulkadyrov

Russian Scientific Research Institute of Hematology and Transfusiology, 16 2-ya Sovetskaya str., Saint Petersburg, Russian Federation, 191024

For correspondence: Sergei Vladimirovich Voloshin, PhD, 16 2-ya Sovetskaya str., Saint Petersburg, Russian Federation, 191024; Tel.: +7(812)274-37-70; e-mail: kuvshinovmd@gmail.com

For citation: Kuvshinov AYu, Voloshin SV, Martynkevich IS, et al. Chronic Lymphocytic Leukemia: Prognostic Significance of Minimal Residual Disease and Potential of Modern Methods of Its Diagnosis and Therapy (Literature Review). Clinical oncohematology. 2016;9(2):191–8 (In Russ).

DOI: 10.21320/2500-2139-2016-9-2-191-198

ABSTRACT

Achieving a complete remission (CR) in patients with chronic lymphocytic leukemia (CLL) has become a feasible goal directly correlating with a prolonged survival. However, a certain number of tumor cells may be present in the patient’s body even when CR has been achieved, and this phenomenon is called a minimal residual disease (MRD). A lot of data confirming the necessity of MRD diagnosing and monitoring has emerged recently, since the MRD has a significant impact on the prognosis of CLL. Achieving MRD-negative remission is an independent predictor of long-term progression-free survival and overall survival. The occurrence of new diagnostic techniques has allowed to define the MRD and to develop standards for its assessment. This paper presents an overview of literature data about MRD, methods of its evaluation, prognostic significance, as well as the methods of eradication.

Keywords: chronic lymphocytic leukemia, minimal residual disease, flow cytometry.

Received: January 5, 2016

Accepted: January 10, 2016

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REFERENCES

  1. Hallek M, Cheson BD, Catovsky D, et al. Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood. 2008;111(12):5446–56. doi: 10.1182/blood-2007-06-093906.
  2. Cave H, van der Werff ten Bosch J, Suciu S, et al. Clinical significance of minimal residual disease in childhood acute lymphoblastic leukemia. European Organization for Research and Treatment of Cancer—Childhood Leukemia Cooperative Group. N Engl J Med. 1998;339(9):591–8. doi: 10.1056/nejm199808273390904.
  3. Andersen NS, Pedersen LB, Laurell A, et al. Preemptive treatment with rituximab of molecular relapse after autologous stem cell transplantation in mantle cell lymphoma. J Clin Oncol. 2009;27(26):4365–70. doi: 10.1200/JCO.2008.21.3116.
  4. Grimwade D, Lo Coco F. Acute promyelocytic leukemia: a model for the role of molecular diagnosis and residual disease monitoring in directing treatment approach in acute myeloid leukemia. Leukemia. 2002;16(10):1959–73. doi: 10.1038/sj.leu.2402721.
  5. Vora A, Goulden N, Mitchell C, et al. Augmented post-remission therapy for a minimal residual disease-defined high-risk subgroup of children and young people with clinical standard-risk and intermediate-risk acute lymphoblastic leukaemia (UKALL 2003): a randomised controlled trial. Lancet Oncol. 2014;15(8):809–18. doi: 10.1016/S1470-2045(14)70243-8.
  6. Moreton P, Kennedy B, Lucas G, et al. Eradication of minimal residual disease in B-cell chronic lymphocytic leukemia after alemtuzumab therapy is associated with prolonged survival. J Clin Oncol. 2005;23(13):2971–9. doi: 10.1200/jco.2005.04.021.
  7. Ritgen M, Bottcher S, Stilgenbauer S, et al. Quantitative MRD monitoring identifies distinct GVL response patterns after allogeneic stem cell transplantation for chronic lymphocytic leukemia: results from the GCLLSG CLL3X trial. Leukemia. 2008;22(7):1377–86. doi: 10.1038/leu.2008.96.
  8. Del Poeta G, Del Principe MI, Buccisano F, et al. Consolidation and maintenance immunotherapy with rituximab improve clinical outcome in patients with B-cell chronic lymphocytic leukemia. Cancer. 2008;112(1):119–28. doi: 10.1002/cncr.23144.
  9. Rawstron AC, Kennedy B, Moreton P, et al. Early prediction of outcome and response to alemtuzumab therapy in chronic lymphocytic leukemia. Blood. 2004;103(6):2027–31. doi: 10.1182/blood-2002-10-3270.
  10. Bosch F, Ferrer A, Villamor N, et al. Fludarabine, cyclophosphamide, and mitoxantrone as initial therapy of chronic lymphocytic leukemia: high response rate and disease eradication. Clin Cancer Res. 2008;14(1):155–61. doi: 10.1158/1078-0432.CCR-07-1371.
  11. Kay NE, Geyer SM, Call TG, et al. Combination chemoimmunotherapy with pentostatin, cyclophosphamide, and rituximab shows significant clinical activity with low accompanying toxicity in previously untreated B chronic lymphocytic leukemia. Blood. 2007;109(2):405–11. doi: 10.1182/blood-2006-07-033274.
  12. Ritgen M, Lange A, Stilgenbauer S, et al. Unmutated immunoglobulin variable heavy-chain gene status remains an adverse prognostic factor after autologous stem cell transplantation for chronic lymphocytic leukemia. Blood. 2003;101(5):2049–53. doi: 10.1182/blood-2002-06-1744.
  13. Hillmen P, Skotnicki AB, Robak T, et al. Alemtuzumab compared with chlorambucil as first-line therapy for chronic lymphocytic leukemia. J Clin Oncol. 2007;25(35):5616–23. doi: 10.1200/jco.2007.12.9098.
  14. Robertson LE, Huh YO, Butler JJ, et al. Response assessment in chronic lymphocytic leukemia after fludarabine plus prednisone: clinical, pathologic, immunophenotypic, and molecular analysis. Blood. 1992;80:29–36.
  15. O’Brien SM, Kantarjian HM, Cortes J, et al. Results of the fludarabine and cyclophosphamide combination regimen in chronic lymphocytic leukemia. J Clin Oncol. 2001;19:1414–20.
  16. Tam CS, O’Brien S, Wierda W, et al. Long-term results of the fludarabine, cyclophosphamide, and rituximab regimen as initial therapy of chronic lymphocytic leukemia. Blood. 2008;112(4):975–80. doi: 10.1182/blood-2008-02-140582.
  17. Robak T, Blonski JZ, Gora-Tybor J, et al. Cladribine alone and in combination with cyclophosphamide or cyclophosphamide plus mitoxantrone in the treatment of progressive chronic lymphocytic leukemia: report of a prospective, multicenter, randomized trial of the Polish Adult Leukemia Group (PALG CLL2). Blood. 2006;108(2):473–9. doi: 10.1182/blood-2005-12-4828.
  18. Moreno C, Villamor N, Colomer D, et al. Clinical significance of minimal residual disease, as assessed by different techniques, after stem cell transplantation for chronic lymphocytic leukemia. Blood. 2006;107(11):4563–9. doi: 10.1182/blood-2005-09-3634.
  19. Milligan DW, Fernandes S, Dasgupta R, et al. Results of the MRC pilot study show autografting for younger patients with chronic lymphocytic leukemia is safe and achieves a high percentage of molecular responses. Blood. 2005;105(1):397–404. doi: 10.1182/blood-2004-01-0298.
  20. Bottcher S, Ritgen M, Pott C, et al. Comparative analysis of minimal residual disease detection using four-color flow cytometry, consensus IgH-PCR, and quantitative IgH PCR in CLL after allogeneic and autologous stem cell transplantation. Leukemia. 2004;18(10):1637–45. doi: 10.1038/sj.leu.2403478.
  21. Rawstron AC, Kennedy B, Evans PA, et al. Quantitation of minimal disease levels in chronic lymphocytic leukemia using a sensitive flow cytometric assay improves the prediction of outcome and can be used to optimize therapy. Blood. 2001;98(1):29–35. doi: 10.1182/blood.v98.1.29.
  22. Maloum K, Sutton L, Baudet S, et al. Novel flow-cytometric analysis based on BCD5+ subpopulations for the evaluation of minimal residual disease in chronic lymphocytic leukaemia. Br J Haematol. 2002;119(4):970–5. doi: 10.1046/j.1365-2141.2002.03956.x.
  23. Никитин Е.А. Дифференцированная терапия хронического лимфолейкоза: Дис. ¼ д-ра мед. наук. М., 2014. 203 с.
    [Nikitin EA. Differentsirovannaya terapiya khronicheskogo limfoleikoza. (Differentiated therapy of chronic lymphocytic leukemia.) [dissertation] Moscow; 2014. 203 p. (In Russ)]
  24. Ripolles L, Ortega M, Ortuno F, et al. Genetic abnormalities and clinical outcome in chronic lymphocytic leukemia. Cancer Genet Cytogenet. 2006;171(1):57–64. doi: 10.1016/j.cancergencyto.2006.07.006.
  25. Van Dongen JJ, Langerak AW, Bruggemann M, et al. Design and standardization of PCR primers and protocols for detection of clonal immunoglobulin and T-cell receptor gene recombinations in suspect lymphoproliferations: report of the BIOMED-2 Concerted Action BMH4-CT98-3936. Leukemia. 2003;17(12):2257–317. doi: 10.1038/sj.leu.2403202.
  26. Rawstron AC, Villamor N, Ritgen M, et al. International standardized approach for flow cytometric residual disease monitoring in chronic lymphocytic leukaemia. Leukemia. 2007;21(5):956–64. doi: 10.1038/sj.leu.2404584.
  27. Keating MJ, O’Brien S, Albitar M, et al. Early results of a chemoimmunotherapy regimen of fludarabine, cyclophosphamide, and rituximab as initial therapy for chronic lymphocytic leukemia. J Clin Oncol. 2005;23(18):4079–88. doi: 10.1200/jco.2005.12.051.
  28. Byrd JC, Peterson BL, Morrison VA, et al. Randomized phase 2 study of fludarabine with concurrent versus sequential treatment with rituximab in symptomatic, untreated patients with B-cell chronic lymphocytic leukemia: Results from Cancer and Leukemia Group B 9712 (CALGB 9712). Blood. 2003;101(1):6–14. doi: 10.1182/blood-2002-04-1258.
  29. Eichhorst BF, Busch R, Hopfinger G, et al. Fludarabine plus cyclophosphamide versus fludarabine alone in first-line therapy of younger patients with chronic lymphocytic leukemia. Blood. 2006;107(3):885–91. doi: 10.1182/blood-2005-06-2395.
  30. Cheson BD, Bennett JM, Grever M, et al. National Cancer Institute-sponsored Working Group guidelines for chronic lymphocytic leukemia: revised guidelines for diagnosis and treatment. Blood. 1996;87:4990–7.
  31. Dreger P, Ritgen M, Bottcher S, et al. The prognostic impact of minimal residual disease assessment after stem cell transplantation for chronic lymphocytic leukemia: Is achievement of molecular remission worthwhile? Leukemia. 2005;19(7):1135–8. doi: 10.1038/sj.leu.2403800.
  32. Wendtner CM, Ritgen M, Schweighofer CD, et al. Consolidation with alemtuzumab in patients with chronic lymphocytic leukemia (CLL) in first remission–experience on safety and efficacy within a randomized multicenter phase III trial of the German CLL Study Group (GCLLSG). Leukemia. 2004;18(6):1093–101. doi: 10.1038/sj.leu.2403354.
  33. Montillo M, Tedeschi A, Miqueleiz S, et al. Alemtuzumab as consolidation after a response to fludarabine is effective in purging residual disease in patients with chronic lymphocytic leukemia. J Clin Oncol. 2006;24(15):2337–42. doi: 10.1200/jco.2005.04.6037.
  34. Rawstron AC, de Tute R, Jack AS, et al. Flow cytometric protein expression profiling as a systematic approach for developing disease-specific assays: identification of a chronic lymphocytic leukaemia-specific assay for use in rituximab-containing regimens. Leukemia. 2006;20(12):2102–10. doi: 10.1038/sj.leu.2404416.
  35. Hallek M, Fingerle-Rowson G, Fink A, et al. Immunochemotherapy with fludarabine (F), cyclophosphamide (C), and rituximab (R) (FCR) versus fludarabine and cyclophosphamide (FC) improves response rates and progression-free survival (PFS) of previously untreated patients (pts) with advanced chronic lymphocytic leukemia (CLL). Blood. 2008;112: Abstract 325.
  36. Wierda W, O’Brien S, Wen S, et al. Chemoimmunotherapy with fludarabine, cyclophosphamide, and rituximab for relapsed and refractory chronic lymphocytic leukemia. J Clin Oncol. 2005;23(18):4070–8. doi: 10.1200/jco.2005.12.516.
  37. Bottcher S, Stilgenbauer S, Busch R, et al: Standardized MRD flow and ASO IGH RQ-PCR for MRD quantification in CLL patients after rituximab-containing immunochemotherapy: A comparative analysis. Leukemia. 2009;23(11):2007–17. doi: 10.1038/leu.2009.140.
  38. Ringelstein-Harlev S, Fineman R. Minimal Residual Disease Surveillance in Chronic Lymphocytic Leukemia by Fluorescence-Activated Cell Sorting. Rambam Maimonides Med J. 2014;5(4): e0027. doi: 10.5041/RMMJ.10161.
  39. Bottcher S, Ritgen M, Fischer K, et al. Minimal residual disease quantification is an independent predictor of progression-free and overall survival in chronic lymphocytic leukemia: a multivariate analysis from the randomized GCLLSG CLL8 trial. J Clin Oncol. 2012;30(9):980–8. doi: 10.1200/JCO.2011.36.9348.
  40. Kovacs G, Bottcher S, Bahlo J, et al. Value of minimal residual disease (MRD) negative status at response evaluation in chronic lymphocytic leukemia (CLL): combined analysis of two phase III studies of the German CLL Study Group (GCLLSG). ASH Annual Meeting Abstracts. 2014: Abstract 23.
  41. Eichhorst B, Fink AM, Busch R, et al. Frontline chemoimmunotherapy with fludarabine (F), cyclophosphamide (C), and rituximab (R) (FCR) shows superior efficacy in comparison to bendamustine (B) and rituximab (BR) in previously untreated and physically fit patients (pts) with advanced chronic lymphocytic leukemia (CLL): Final analysis of an international, randomized study of the German CLL Study Group (GCLLSG) (CLL10 study). Proc ASH 2014: Abstract 19.
  42. Garifullin A, Kuvshinov A, Voloshin S, et al. The frequency of occurrence of minimal residual disease (MRD) into different prognostic groups of patients with chronic lymphocytic leukemia (CLL). Intern Hematol Club. [Internet] 2015 Nov 6–7 [cited 2016 April 18] Available from: http://www.comtecmed.com/IHC/2015/poster_list.aspx.
  43. Schweighofer CD, Ritgen M, Eichhorst BF, et al. Consolidation with alemtuzumab improves progression-free survival in patients with chronic lymphocytic leukaemia (CLL) in first remission: long-term follow-up of a randomized phase III trial of the German CLL Study Group (GCLLSG). Br J Haematol. 2009;144(1):95–8. doi: 10.1111/j.1365-2141.2008.07394.x.
  44. Wiernik PH, Adiga GU. Single-agent rituximab in treatment-refractory or poor prognosis patients with chronic lymphocytic leukemia. Curr Med Res Opin. 2011;27(10):1987–93. doi: 10.1185/03007995.2011.615307.
  45. Abrisqueta P, Villamor N, Terol MJ, et al. Rituximab maintenance after first-line therapy with rituximab, fludarabine, cyclophosphamide, and mitoxantrone (R-FCM) for chronic lymphocytic leukemia. Blood. 2013;122(24):3951–9. doi: 10.1182/blood-2013-05-502773.
  46. Shanafelt TD, Ramsay AG, Zent CS, et al. Long-term repair of T-cell synapse activity in a phase II trial of chemoimmunotherapy followed by lenalidomide consolidation in previously untreated chronic lymphocytic leukemia (CLL). Blood. 2013;121(20):4137–41. doi: 10.1182/blood-2012-12-470005.
  47. Dreger P, Dohner H, Ritgen M, et al. Allogeneic stem cell transplantation provides durable disease control in poor-risk chronic lymphocytic leukemia: long-term clinical and MRD results of the German CLL Study Group CLL3X trial. Blood. 2010;116(14)2438–47. doi: 10.1182/blood-2010-03-275420.
  48. Byrd JC, Furman RR, Coutre SE, et al. Targeting BTK with ibrutinib in relapsed chronic lymphocytic leukemia. N Engl J Med. 2013;369(1):32–42. doi: 10.1056/NEJMoa1215637.
  49. O’Brien S, Furman RR, Coutre SE, et al. Ibrutinib as initial therapy for elderly patients with chronic lymphocytic leukaemia or small lymphocytic lymphoma: an open-label, multicentre, phase 1b/2 trial. Lancet Oncol. 2014;15(1):48–58. doi: 10.1016/S1470-2045(13)70513-8.
  50. Furman RR, Sharman JP, Coutre SE, et al. Idelalisib and rituximab in relapsed chronic lymphocytic leukemia. N Engl J Med. 2014;370(11):997–1007. doi: 10.1056/NEJMoa1315226.
  51. O’Brien S, Lamanna N, Kipps TJ, et al. Update on a Phase 2 Study of Idelalisib in Combination with Rituximab in Treatment-Naive Patients ³65 Years with Chronic Lymphocytic Leukemia (CLL) or Small Lymphocytic Lymphoma (SLL). Blood. 2014;124: Abstract 1994.

 

Allogeneic Hematopoietic Stem Cell Transplantation in Myelodysplastic Syndromes and Clinical Significance of WT1 Gene Overexpression

MYELOID TUMORS , , , , ,

N.N. Mamaev1, A.V. Gorbunova1, T.L. Gindina1, E.V. Morozova1, Ya.V. Gudozhnikova1, O.A. Slesarchuk1, V.N. Ovechkina1, A.A. Rats1, E.G. Boichenko2, E.A. Ukrainchenko3, V.M. Kravtsova1, A.V. Evdokimov1, I.M. Barkhatov1, S.N. Bondarenko1, B.V. Afanas’ev1

1 R.M. Gorbacheva Scientific Research Institute of Pediatric Hematology and Transplantation; Academician I.P. Pavlov First St. Petersburg State Medical University, 12 Rentgena str., Saint Petersburg, Russian Federation, 197022

2 Municipal Children’s Hospital No. 1, 14 Avangardnaya str., Saint Petersburg, Russian Federation, 198205

3 Alexandrovskaya Municipal Hospital No. 17, 4 pr-t Solidarnosti, Saint Petersburg, Russian Federation, 193312

For correspondence: N.N. Mamaev, DSci, Professor, 12 Rentgena str., Saint Petersburg, Russian Federation, 197022; Tel: +7(812)233-12-43; e-mail: nikmamaev524@gmail.com

For citation: Mamaev N.N., Gorbunova A.V., Gindina T.L., Morozova E.V., Gudozhnikova Ya.V., Slesarchuk O.A., Ovechkina V.N., Rats A.A., Boichenko E.G., Ukrainchenko E.A., Kravtsova V.M., Evdokimov A.V., Barkhatov I.M., Bondarenko S.N., Afanas’ev B.V. Allogeneic Hematopoietic Stem Cell Transplantation in Myelodysplastic Syndromes and Clinical Significance of WT1 Gene Overexpression. Klin. Onkogematol. 2014; 7(4): 551–563 (In Russ.).

ABSTRACT

The results of allogeneic hematopoietic stem cell transplantation (HSCT) in 17 patients (pts, 11 male, 6 female) with myelodysplastic syndromes (3 RA/RARS/RCMD, 5 RAEB-1, 7 RAEB-2, 2 JMML) are presented. The median age was 26 years with a range between 1 and 55 years. Serial cytogenetic investigations were carried out in all of them. Seven pts demonstrated monosomy 7, which was associated with other chromosome abnormalities in 4 cases. In addition, deletion at 11q23 (n = 3), trisomy 8 (n = 2) and 21 (n = 2), involvement into rearrangement at 3q (n = 2), t(6;9) translocation, and others more rare abnormalities were found. Prior to aHSCT, 11 of 7 received hypomethylating agents (HA) which proved to be effective in a half of them. In order to prepare for aHSCT, ablative (busulfan, cyclophosphamide) or non-ablative (fludarabine, cyclophosphamide) conditioning regimes were applied (4 and 13 respectively). Repeated aHSCT was carried out in 6 pts because of transplant rejection or post-transplant relapses. Molecular monitoring of minimal residual disease as well as early diagnosis of these relapses was performed by means of serial tests of the WT1 gene level expression and donor chimerism. Maximum WT1 values varied between 15 and 43133 copies/104 copies of ABL gene; and molecular relapses were registered in a half of them, including 5 patients with transformation into acute leukemia (AL). HA were used for prevention and treatment of relapses in 4 (24 %) patients; and HA were combined with donor lymphocyte infusions. Standard chemotherapy was applied for these purposes relatively rarely. This study demonstrated WT1 gene overexpression to be not only an important marker for diagnosis of post-transplant MDS/AL relapses, but it also can be used for evaluation of the treatment efficacy.

Keywords: myelodysplastic syndromes, allogeneic HSCT, post-transplant relapses, minimal residual disease, molecular monitoring, serial WT1 gene expression.

Accepted: September 30, 2014

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REFERENCES 

  1. Barrett A.J., Battiwala M. Relapse after allogeneic stem cell transplantation. Exp. Rev. Hematol. 2012; 3(4): 429–41.
  2. Tamura K., Kanazawa T., Suzuki M. et al. Successful rapid discontinuation of immunosuppressive therapy at molecular relapse after allogeneic bone marrow transplantation in a pediatric patient with myelodysplastic syndrome. Am. J. Hematol. 2006; 81: 139–41.
  3. Wertheim G.B., Bagg A. Minimal residual disease testing to predict relapse following transplant for AML and high-grade myelodysplastic syndromes. Exp. Rev. Mol. Diagn. 2011; 11(4): 361–6.
  4. Brieger J., Weidmann E., Fenchel K. et al. The expression of the Wilms’ tumor gene in acute myelocytic leukemias as a possible marker for leukemic blast cells. Leukemia. 1994; 8: 2138–43.
  5. Inoue K., Sugiyama H., Ogawa H. et al. WT1 as a new prognostic factor and a new marker for the detection of minimal residual disease in acute leukemia. Blood. 1994; 84: 3071–9.
  6. Inoue K., Ogawa H., Yamagami T. et al. Long-term follow-up of minimal residual disease in leukemia patients by monitoring WT1 (Wilms tumor gene) expression levels. Blood. 1996; 88: 2267–78.
  7. Tamaki H., Ogawa H., Inoue K. et al. Increased expression of the Wilms tumor gene (WT1) at relapse in acute leukemia. Blood. 1996; 88: 4396–8.
  8. Patmasirivat P., Fraizer G., Kantarjian H. et al. WT1 and GATA1 expression in myelodysplastic syndrome and acute leukemia. Leukemia. 1999; 13: 891–900.
  9. Ogawa H., Ikegame K., Kawakami M., Tamaki H. WT1 gene transcript assay for relapse in acute leukemia after transplantation. Leuk. Lymphoma. 2004; 45: 1747–53.
  10. Cilloni D., Gottardi E., De Micheli D. et al. Quantitative assessment of WT1 expression by real time quantitative PCR may be a useful tool for monitoring minimal residual disease in acute leukemia patients. Leukemia. 2002; 16: 2115–21.
  11. Cilloni D., Messa F., Arruga F. et al. Early prediction of treatment outcome in acute myeloid leukemia by measurement of WT1 transcript levels in peripheral blood samples collected after chemotherapy. Haematologica. 2008; 93: 921–4.
  12. Candoni A., Tribelli M., Cilloni D. et al. Quantitative assessment of WT1 gene expression after allogeneic stem cell transplantation is a useful tool for monitoring minimal residual disease in acute myeloid leukemia. Eur. J. Haematol. 2009; 82: 61–8.
  13. Miyawaki S., Hatsumi N., Tamaki T. et al. Prognostic potential of detection of WT1 mRNA level in peripheral blood in adult acute myeloid leukemia. Leuk. Lymphoma 2010; 51: 1855–61.
  14. Zhao X.-S., Jin S., Zhu H.-H. et al. Wilms’ tumor gene 1 expression: an independent acute leukemia prognostic indicator following allogeneic hematopoietic SCT. Bone Marrow Transplant. 2012; 47: 499–507.
  15. Nomdedeu J.F., Hoyos M., Carricondo M. et al. Bone marrow WT1 levels at diagnosis, post-induction and post-intensification in adult de novo AML. Leukemia. 2013; 27: 2157–64.
  16. Pozzi S., Geroldi S., Tedone E. et al. Leukaemia relapse after allogeneic transplants for acute myeloid leukaemia: predictive role of WT1 expression. Br. J. Haematol. 2013, 160: 503–9.
  17. Frairia Ch., Aydin S., Riera L. et al. WT1 expression in acute myeloid leukemia: a useful marker for improving therapy response evaluation. Blood. 2013; 123(21): 2588.
  18. Tamaki H., Ogawa H., Ohyashiki K. et al. The Wilm’s tumor gene is a good marker for diagnosis of disease progression of myelodysplastic syndromes. Leukemia. 1999; 13: 393–9.
  19. Patmasiriwat P., Fraizer G., Kantarjian H. et al. WT1 and GATA1 expression in myelodysplastic syndrome and acute leukemia. Leukemia. 1999; 13: 891–900.
  20. Cilloni D., Gottardi E., Messa F. et al. Significant correlation between the degree of WT1 expression and the international prognostic scoring system score in patients with myelodysplastic syndromes. J. Clin. Oncol. 2003; 21: 1988–95.
  21. Cilloni D., Saglio G. WT1 as a universal marker for minimal residual disease detection and quantification in myeloid leukemias and in myelodysplastic syndrome. Acta Haematologica. 2004; 112: 79–84.
  22. Абдулкадыров К.М., Грицаев С.В., Капустин С.И. и др. Экспрессия гена опухоли Вилмса (WT1) в клетках крови больных миелодиcпластическим синдромом. Вопросы онкологии 2004; 50(6): 668–71. [Abdulkadyrov K.M., Gritsaev S.V., Kapustin S.I. et al. Wilms tumor gene (WT1) expression in blood cells of patients with myelodysplastic syndrome. Voprosy Onkologii. 2004; 50(6): 668–71. (In Russ.)]
  23. Bader P., Niemeyer C., Weber G. et al. WT1 gene expression: useful marker for minimal residual disease in childhood myelodysplastic syndromes and juvenile myelomonocytic leukemia? Eur. J. Haematol. 2004; 73: 25–8.
  24. Iwasaki T., Sugisaki C., Nagata K. et al. Wilms’ tumor 1 message and protein expression in bone marrow failure syndrome and acute leukemia. Pathol. Int. 2007; 57: 645–51.
  25. Qin Y.-Z., Zhu H.-H., Liu Y.-R. et al. PRAME and WT1 transcripts constitute a good molecular marker combination for monitoring minimal residual disease in myelodysplastic syndromes. Leuk. Lymphoma. 2012; DOI: 10.3109/10428194.2012.743656.
  26. Ueda Y., Mizutani C., Nannya Y. et al. Clinical evaluation of WT1 mRNA expression levels in peripheral blood and bone marrow in patients with myelodysplastic syndromes. Leuk. Lymphoma. 2013; 54(7): 1450–8.
  27. Lange T., Hubmann M., Burkhardt R. et al. Monitoring of WT1 expression in PB and CD34+ donor chimerism of BM predicts early relapse in AML and MDS patients after hematopoietic cell transplantation with reduced-intensity conditioning. Leukemia. 2011; 25: 498–505.
  28. Maurer U., Brieger J., Weidmann E. et al. The Wilms’ tumor gene is expressed in a subset of CD34+ progenitors and downregulated early in the course of differentiation in vitro. Exp. Hematol. 1997; 25: 945–50.
  29. Kwon M., Marti nez-Laperche C., Infante M. et al. Evaluation of minimal residual disease by real-time quantitative PCR of Wilms’ Tumor 1 expression in patients with acute myelogenous leukemia after allogeneic stem cell transplantation: correlation with flow cytometry and chimerism. Biol. Blood Marrow Transplant. 2012; 18: 1235–42.
  30. Jacobsohn D.A., Tse W.T., Chaleff S. et al. High WT1 gene expression before haematopoietic stem cell transplant in children with acute myeloid leukaemia predicts poor event-free survival. Br. J. Haematol. 2009; 146: 669–74.
  31. Мамаев Н.Н., Горбунова А.В., Гиндина Т.Л. и др. Лейкозы и миело- диспластические синдромы с высокой экспрессией гена EVI1: теоретиче- ские и клинические аспекты. Клин. онкогематол. 2012; 5(4): 361–4. [Mamaev N.N., Gorbunova A.V., Gindina T.L. et al. Leukemias and myelodysplastic syndromes with high EVI1 gene expression: theoretical and clinical aspects. Klin. Onkogematol. 2012; 5(4): 361–4. (In Russ.)]
  32. Alonso-Dominguez J.M., Tenorio M., Velasco D. et al. Correlation of WT1 expression with the burden of total and residual leukemic blasts in bone marrow samples of acute myeloid leukemia patients. Cancer Gen. 2012; 205: 190–1.
  33. Gerds A.T., Deeg H.J. Transplantation for myelodysplastic syndrome in the era of hypomethylating agents. Curr. Opin. Hematol. 2012; 19: 71–5.
  34. Nishihori T., Perkins J., Mishra A. et al. Pretransplantation 5-Azacitidine in high-risk myelodysplastic syndrome. Biol. Blood Marrow Transplant. 2014; 20: 776–80.
  35. Raza A., Gezer S., Mundle S. et al. Apoptosis in bone marrow biopsy samples involving stromal and hematopoietic cells in 50 patients with myelodysplastic syndromes. Blood. 1995; 86(1): 268–76.