bloodjourГематология и трансфузиологияRussian journal of hematology and transfusiology0234-57302411-3042ООО Издательский дом «Практика»10.35754/0234-5730-2026-71-1-46-59bloodjour-727Research ArticleОРИГИНАЛЬНЫЕ СТАТЬИORIGINAL ARTICLESМинорные клоны с внутренней тандемной дупликацией гена FLT3 при NPM1-позитивном остром миелоидном лейкозеMinor clones with internal tandem duplication of the FLT3 gene in NPM1-positive acute myeloid leukemiahttps://orcid.org/0000-0003-1936-0084СидороваЮ. В.SidorovaYu. V.

Сидорова Юлия Владимировна, кандидат медицинских наук, врач клинической лабораторной диагностики лаборатории молекулярной гематологии

125167, г. Москва 

Yulia V. Sidorova, Cand. Sci. (Med.), Doctor of Clinical Laboratory Diagnostics, Laboratory of Molecular Hematology

125167, Moscow 

sidorova.y@blood.ruhttps://orcid.org/0009-0005-2774-1938КостроминаМ. А.KostrominaM. A.

Костромина Мария Александровна, студентка 

119991, г. Москва

Maria A. Kostromina, student  

119991, Moscow 

mariya.kostromina.01@mail.ruhttps://orcid.org/0000-0002-7036-9968СеверинаН. А.SeverinaN. A.

Северина Наталия Александровна, кандидат биологических наук, врач клинической лабораторной диагностики лаборатории молекулярной гематологии

125167, г. Москва 

Nataliya A. Severina, Cand. Sci. (Biol.), Doctor of Clinical Laboratory Diagnostics, Laboratory of Molecular Hematology

125167, Moscow 

severina.n@blood.ruhttps://orcid.org/0000-0002-6253-3334БидерманБ. В.BidermanB. V.

Бидерман Белла Вениаминовна, кандидат биологических наук, старший научный сотрудник лаборатории молекулярной гематологии

125167, г. Москва 

Bella V. Biderman, Cand. Sci. (Biol.), Senior Researcher, Laboratory of Molecular Hematology

125167, Moscow 

biderman.b@blood.ruhttps://orcid.org/0000-0001-8044-598XЧабаеваЮ. А.ChabaevaYu. A.

Чабаева Юлия Александровна, кандидат технических наук, заместитель начальника информационно-аналитического отдела

125167, г. Москва 

Yulia A. Chabaeva, Cand. Sci. (Tech.), Deputy Head of the Informational Analytical Department

125167, Moscow 

chabaeva.y@blood.ruhttps://orcid.org/0000-0002-6288-7570КуликовС. М.KulikovS. M.

Куликов Сергей Михайлович, кандидат технических наук, начальник информационно-аналитического отдела

125167, г. Москва 

Sergey M. Kulikov, Cand. Sci. (Tech.), Head of the Informational Analytical Department

125167, Moscow 

kulikov.s@blood.ruhttps://orcid.org/0000-0002-3548-8929КашлаковаА. И.KashlakovaA. I.

Кашлакова Анастасия Игоревна, гематолог отделения химиотерапии гемобластозов и депрессий кроветворения с блоком трансплантации костного мозга и гемопоэтических стволовых клеток

125167, г. Москва 

Anastasia I. Kashlakova, Hematologist, Department of Chemotherapy of Hemoblastosis and Hematopoietic Depressions with Bone marrow and Hematopoietic Stem Cell Transplantation Unit

125167, Moscow 

kashlakova.a@blood.ruhttps://orcid.org/0009-0002-7001-0493ДробининаА. П.DrobininaA. P.

Дробинина Анна Павловна, гематолог дневного стационара онкологии и химиотерапии гемобластозов и депрессий кроветворения 

125167, г. Москва 

Anna P. Drobinina, Hematologist of the Department of Chemotherapy of Hemoblastosis and Hematopoietic Depressions

125167, Moscow 

drobinina.a@blood.ruhttps://orcid.org/0000-0002-8337-2242ЛукьяноваИ. А.LukianovaI. A.

Лукьянова Ирина Анатольевна, кандидат медицинских наук, заведующая дневным стационаром онкологии и химиотерапии гемобластозов и депрессий кроветворения

125167, г. Москва 

Irina A. Lukianova, Cand. Sci. (Med.), Head of the Day clinic of Chemotherapy of Hemoblastosis and Hematopoietic Depressions

125167, Moscow 

lukyanova.i@blood.ruhttps://orcid.org/0000-0001-9463-9187СудариковА. Б.SudarikovA. B.

Судариков Андрей Борисович, доктор биологических наук, заведующий лабораторией молекулярной гематологии

125167, г. Москва 

Andrey B. Sudarikov, Dr. Sci. (Biol.), Head of the Laboratory of Molecular Hematology

125167, Moscow 

dusha@blood.ruhttps://orcid.org/0000-0001-6177-3566ПаровичниковаЕ. Н.ParovichnikovaE. N.

Паровичникова Елена Николаевна, доктор медицинских наук, член-корреспондент РАН, генеральный директор 

125167, г. Москва 

Elena N. Parovichnikova, Dr. Sci. (Med.), Corresponding Member of the RAS, CEO

125167, Moscow 

parovichnikova.e@blood.ruФГБУ «Национальный медицинский исследовательский центр гематологии» Министерства здравоохранения Российской ФедерацииРоссияNational Medical Research Center for HematologyRussian FederationФакультет фундаментальной медицины МНОИ ФГБОУ ВО «Московский государственный университет им. М. В. Ломоносова»РоссияFaculty of Fundamental Medicine MREI, M.V. Lomonosov Moscow State UniversityRussian Federation2026070420267114659Copyright © Сидорова Ю.В., Костромина М.А., Северина Н.А., Бидерман Б.В., Чабаева Ю.А., Куликов С.М., Кашлакова А.И., Дробинина А.П., Лукьянова И.А., Судариков А.Б., Паровичникова Е.Н., 20262026Сидорова Ю.В., Костромина М.А., Северина Н.А., Бидерман Б.В., Чабаева Ю.А., Куликов С.М., Кашлакова А.И., Дробинина А.П., Лукьянова И.А., Судариков А.Б., Паровичникова Е.Н.Sidorova Y.V., Kostromina M.A., Severina N.A., Biderman B.V., Chabaeva Y.A., Kulikov S.M., Kashlakova A.I., Drobinina A.P., Lukianova I.A., Sudarikov A.B., Parovichnikova E.N.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.htjournal.ru/jour/article/view/727Введение

Введение. Внутренние тандемные дупликации (internal tandem duplication, ITD) в гене FMS-подобной тирозинкиназы 3 (FLT3) — фактор неблагоприятного прогноза при остром миелоидном лейкозе (ОМЛ). Своевременное обнаружение FLT3-ITD позволяет определить группу риска ОМЛ и назначить таргетные препараты.

Цель

Цель: оценить частоту выявления минорных клонов FLT3-ITD и их ассоциацию с прогнозом и динамикой минимальной остаточной болезни у больных с NPM1-позитивным ОМЛ.

Материалы и методы

Материалы и методы. В исследование включены 58 больных ОМЛ, относившихся к промежуточной или благоприятной группам риска, имевших мутацию гена NPM1. Больным проводили терапию по протоколу «ОМЛ-21». Молекулярно-генетическую диагностику мутаций FLT3 проводили методами полимеразной цепной реакции (ПЦР) с фрагментным анализом (ПЦР-ФА), ПЦР-ФА с двойной меткой и методом ПЦР с тандемной дупликацией (ТД-ПЦР), мутаций NPM1 — методами ПЦР-ФА (первичная диагностика), аллель-специфичной ПЦР (для оценки минимальной остаточной болезни (МОБ)).

Результаты

Результаты. Применение ПЦР-ФА с двойной меткой и ТД-ПЦР позволило идентифицировать минорные клоны FLT3- ITD у 12 (36 %) из 33 больных, у которых данные клоны не были выявлены стандартным методом, а также выявить дополнительные мутации FLT3-ITD у 52 % (у 13 из 25) «FLT3-позитивных» больных. На исследованной когорте значимой ассоциации наличия минорных клонов FLT3-ITD с динамикой снижения МОБ, частотой достижения МОБ-негативного статуса, общей и безрецидивной выживаемости у больных с NPM1(+) ОМЛ не было обнаружено.

Заключение

Заключение. Ассоциация минорных клонов FLT3-ITD с клиническими особенностями заболевания у больных ОМЛ и мутациями гена NPM1 минимальна или отсутствует. У этих больных не требуется применение дополнительных более чувствительных методов выявления мутаций FLT3-ITD на первичном этапе диагностики и включение ингибиторов тирозинкиназ в программную терапию. Однако окончательные выводы можно будет сделать, если будет показано, что выявление в дебюте минорных клонов с мутацией FLT3-ITD не ассоциировано с возникновением рецидивов с учетом выполняемого протокола лечения.

Introduction

Introduction. Internal tandem duplication (ITD) in the FMS-like tyrosine kinase 3 (FLT3) gene is a poor prognostic factor in acute myeloid leukemia (AML). Identification of a group of such patients raises the question of prescribing targeted therapy to them.

Aim

Aim: to assess the significance of minor FLT3-ITD clones in AML with NPM1 mutation.

Materials and methods

Materials and methods. The study included 58 patients belonging to the intermediate or favorable risk group. Patients underwent therapy according to the AML-21 protocol. Molecular genetic diagnostics of FLT3 mutations was performed by polymerase chain reaction (PCR) with fragment analysis (PCR-FA), PCR-FA with double label and tandem duplication PCR (TD-PCR), NPM1 mutations — by PCR-FA (primary diagnostics) and allele-specific PCR to assess minimal residual disease (MRD).

Results

Results. Using double-labeled PCR-FA and TD-PCR, allowed to identify minor FLT3-ITD clones in 12 (36 %) of 33 patients in whom these clones were not detected by the standard method. We also identified additional FLT3-ITD mutations in 13 (52 %) of 25 FLT3-positive patients. In the studied cohort no significant association of minor FLT3-ITD clones with the dynamics of MRD reduction, the percentage of patients achieving MRD-negative status, overall and relapse-free survival among patients with NPM1(+) AML was found.

Conclusion

Conclusion. The association of minor FLT3-ITD clones with clinical features of the disease in patients with AML and NPM1 gene mutations is minimal or absent. It can be assumed that these patients do not require the use of additional more sensitive methods for detecting FLT3-ITD mutations at the primary stage of diagnosis and the inclusion of tyrosine kinase inhibitors in program therapy. However, final conclusions can be made if it is shown that the detection of minor clones with FLT3-ITD mutation at the onset is not associated with the occurrence of relapses, taking into account the treatment protocol.

острый миелоидный лейкозПЦРFLT3-ITDминорные клоныфрагментный анализкапиллярный электрофорезNPM1МОБacute myeloid leukemiaPCRFLT3-ITDminor clonesfragment analysiscapillar electrophoresisNPM1MRDReferencesKazi J.U., Rönnstrand L. FMS-like Tyrosine Kinase 3/FLT3: From Basic Science to Clinical Implications. Physiol Rev. 2019;99(3):1433–66. DOI: 10.1152/physrev.00029.2018.Kazi J.U., Rönnstrand L. FMS-like Tyrosine Kinase 3/FLT3: From Basic Science to Clinical Implications. Physiol Rev. 2019;99(3):1433–66. DOI: 10.1152/physrev.00029.2018.Grafone T., Palmisano M., Nicci C., Storti S. An overview on the role of FLT3- tyrosine kinase receptor in acute myeloid leukemia: Biology and treatment. Oncol. Rev. 2012;6:e8. DOI: 10.4081/oncol.2012.e8.Grafone T., Palmisano M., Nicci C., Storti S. An overview on the role of FLT3- tyrosine kinase receptor in acute myeloid leukemia: Biology and treatment. Oncol. Rev. 2012;6:e8. DOI: 10.4081/oncol.2012.e8.Kiyoi H., Naoe T. FLT3 in human hematologic malignancies. Leuk Lymphoma. 2002;43(8):1541–7. DOI: 10.1080/1042819021000002866.Kiyoi H., Naoe T. FLT3 in human hematologic malignancies. Leuk Lymphoma. 2002;43(8):1541–7. DOI: 10.1080/1042819021000002866.Schneider F., Hoster E., Schneider S., et al. Age-dependent frequencies of NPM1 mutations and FLT3-ITD in patients with normal karyotype AML (NK-AML) Ann. Hematol. 2012;91:9–18. DOI: 10.1007/s00277-011-1280-6.Schneider F., Hoster E., Schneider S., et al. Age-dependent frequencies of NPM1 mutations and FLT3-ITD in patients with normal karyotype AML (NK-AML) Ann. Hematol. 2012;91:9–18. DOI: 10.1007/s00277-011-1280-6.Ley T.J., Miller C., Ding L., et al. The Cancer Genome Atlas Research Network. Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia. N. Engl. J. Med. 2013;368:2059–74. DOI: 10.1056/nejmoa1301689.Ley T.J., Miller C., Ding L., et al. The Cancer Genome Atlas Research Network. Genomic and epigenomic landscapes of adult de novo acute myeloid leukemia. N. Engl. J. Med. 2013;368:2059–74. DOI: 10.1056/nejmoa1301689.Papaemmanuil E., Gerstung M., Bullinger L., et al. Genomic Classification and Prognosis in Acute Myeloid Leukemia. N. Engl. J. Med. 2016;374:2209–21. DOI: 10.1056/NEJMoa1516192.Papaemmanuil E., Gerstung M., Bullinger L., et al. Genomic Classifi cation and Prognosis in Acute Myeloid Leukemia. N. Engl. J. Med. 2016;374:2209–21. DOI: 10.1056/NEJMoa1516192.Kayser S., Schlenk R.F., Londono M.C., et al. German-Austrian AML Study Group (AMLSG). Insertion of FLT3 internal tandem duplication in the tyrosine kinase domain-1 is associated with resistance to chemotherapy and inferior outcome. Blood. 2009;114(12):2386–92. DOI: 10.1182/blood-2009-03-209999.Kayser S., Schlenk R.F., Londono M.C., et al. German-Austrian AML Study Group (AMLSG). Insertion of FLT3 internal tandem duplication in the tyrosine kinase domain-1 is associated with resistance to chemotherapy and inferior outcome. Blood. 2009;114(12):2386–92. DOI: 10.1182/blood-2009-03-209999.Fröhling S., Schlenk R.F., Breitruck J., et al. Prognostic significance of activating FLT3 mutations in younger adults (16 to 60 years) with acute myeloid leukemia and normal cytogenetics: A study of the AML Study Group Ulm. Blood. 2002;100:4372–80. DOI: 10.1182/blood-2002-05-1440.Fröhling S., Schlenk R.F., Breitruck J., et al. Prognostic signifi cance of activating FLT3 mutations in younger adults (16 to 60 years) with acute myeloid leukemia and normal cytogenetics: A study of the AML Study Group Ulm. Blood. 2002;100:4372–80. DOI: 10.1182/blood-2002-05-1440.Gale R.E., Green C., Allen C., et al. The impact of FLT3 internal tandem duplication mutant level, number, size, and interaction with NPM1 mutations in a large cohort of young adult patients with acute myeloid leukemia. Blood. 2008;111:2776– 84. DOI: 10.1182/blood-2007-08-109090.Gale R.E., Green C., Allen C., et al. The impact of FLT3 internal tandem duplication mutant level, number, size, and interaction with NPM1 mutations in a large cohort of young adult patients with acute myeloid leukemia. Blood. 2008;111:2776– 84. DOI: 10.1182/blood-2007-08-109090.Stone R.M., Mandrekar S.J., Sanford B.L., et al. Midostaurin plus Chemotherapy for Acute Myeloid Leukemia with a FLT3 Mutation. N. Engl. J. Med. 2017;377:454–64. DOI: 10.1056/NEJMoa1614359.Stone R.M., Mandrekar S.J., Sanford B.L., et al. Midostaurin plus Chemotherapy for Acute Myeloid Leukemia with a FLT3 Mutation. N. Engl. J. Med. 2017;377:454–64. DOI: 10.1056/NEJMoa1614359.Perl A.E., Martinelli G., Cortes J.E., et al. Gilteritinib or Chemotherapy for Relapsed or Refractory FLT3-Mutated AML. N. Engl. J. Med. 2019;381:1728–40. DOI: 10.1056/NEJMoa1902688.Perl A.E., Martinelli G., Cortes J.E., et al. Gilteritinib or Chemotherapy for Relapsed or Refractory FLT3-Mutated AML. N. Engl. J. Med. 2019;381:1728–40. DOI: 10.1056/NEJMoa1902688.Badar T., Kantarjian H.M., Nogueras-Gonzalez G.M., et al. Improvement in clinical outcome of FLT3 ITD mutated acute myeloid leukemia patients over the last one and a half decade. Am J Hematol. 2015;90(11):1065–70. DOI: 10.1002/ajh.24140.Badar T., Kantarjian H.M., Nogueras-Gonzalez G.M., et al. Improvement in clinical outcome of FLT3 ITD mutated acute myeloid leukemia patients over the last one and a half decade. Am J Hematol. 2015;90(11):1065–70. DOI: 10.1002/ajh.24140.Oñate G., Pratcorona M., Garrido A. et al. Survival improvement of patients with FLT3 mutated acute myeloid leukemia: results from a prospective 9 years cohort. Blood Cancer J. 2023;13(1):69. DOI: 10.1038/s41408-023-00839-1.Oñate G., Pratcorona M., Garrido A. et al. Survival improvement of patients with FLT3 mutated acute myeloid leukemia: results from a prospective 9 years cohort. Blood Cancer J. 2023;13(1):69. DOI: 10.1038/s41408-023-00839-1.Daver N., Schlenk R.F., Russell N.H., Levis M.J. Targeting FLT3 mutations in AML: Review of current knowledge and evidence. Leukemia. 2019;33:299–312. DOI: 10.1038/s41375-018-0357-9.Daver N., Schlenk R.F., Russell N.H., Levis M.J. Targeting FLT3 mutations in AML: Review of current knowledge and evidence. Leukemia. 2019;33:299–312. DOI: 10.1038/s41375-018-0357-9.Shouval R., Shlush L.I., Yehudai-Resheff S., et al. Single cell analysis exposes intratumor heterogeneity and suggests that FLT3-ITD is a late event in leukemogenesis. Exp Hematol. 2014;42(6):457–63. DOI: 10.1016/j.exphem.2014.01.010.Shouval R., Shlush L.I., Yehudai-Resheff S., et al. Single cell analysis exposes intratumor heterogeneity and suggests that FLT3-ITD is a late event in leukemogenesis. Exp Hematol. 2014;42(6):457–63. DOI: 10.1016/j.exphem.2014.01.010.Onecha E., Rapado I., Luz Morales M., et al. Monitoring of clonal evolution of acute myeloid leukemia identifies the leukemia subtype, clinical outcome and potential new drug targets for post-remission strategies or relapse. Haematologica. 2021;106(9):2325–33. DOI: 10.3324/haematol.2020.254623.Onecha E., Rapado I., Luz Morales M., et al. Monitoring of clonal evolution of acute myeloid leukemia identifi es the leukemia subtype, clinical outcome and potential new drug targets for post-remission strategies or relapse. Haematologica. 2021;106(9):2325–33. DOI: 10.3324/haematol.2020.254623.McCormick S.R., McCormick M.J., Grutkoski P.S., et al. FLT3 mutations at diagnosis and relapse in acute myeloid leukemia: Cytogenetic and pathologic correlations, including cuplike blast morphology. Arch. Pathol. Lab. Med. 2010;134:1143–51. DOI: 10.5858/2009-0292-OA.1McCormick S.R., McCormick M.J., Grutkoski P.S., et al. FLT3 mutations at diagnosis and relapse in acute myeloid leukemia: Cytogenetic and pathologic correlations, including cuplike blast morphology. Arch. Pathol. Lab. Med. 2010;134:1143–51. DOI: 10.5858/2009-0292-OA.1.Tiesmeier J., Müller-Tidow C., Westermann A., et al. Evolution of FLT3-ITD and D835 activating point mutations in relapsing acute myeloid leukemia and response to salvage therapy. Leuk Res. 2004 Oct;28(10):1069–74. DOI: 10.1016/j.leukres.2004.02.009.Tiesmeier J., Müller-Tidow C., Westermann A., et al. Evolution of FLT3-ITD and D835 activating point mutations in relapsing acute myeloid leukemia and response to salvage therapy. Leuk Res. 2004 Oct;28(10):1069–74. DOI: 10.1016/j.leukres.2004.02.009.Cloos J., Goemans B.F., Hess C.J., et al. Stability and prognostic influence of FLT3 mutations in paired initial and relapsed AML samples. Leukemia. 2006;20(7):1217–20. DOI: 10.1038/sj.leu.2404246.Cloos J., Goemans B.F., Hess C.J., et al. Stability and prognostic infl uence of FLT3 mutations in paired initial and relapsed AML samples. Leukemia. 2006;20(7):1217–20. DOI: 10.1038/sj.leu.2404246.Kottaridis P.D., Gale R.E., Langabeer S.E., et al. Studies of FLT3 mutations in paired presentation and relapse samples from patients with acute myeloid leukemia: implications for the role of FLT3 mutations in leukemogenesis, minimal residual disease detection, and possible therapy with FLT3 inhibitors. Blood. 2002;100(7):2393–8. DOI: 10.1182/blood-2002-02-0420.Kottaridis P.D., Gale R.E., Langabeer S.E., et al. Studies of FLT3 mutations in paired presentation and relapse samples from patients with acute myeloid leukemia: implications for the role of FLT3 mutations in leukemogenesis, minimal residual disease detection, and possible therapy with FLT3 inhibitors. Blood. 2002;100(7):2393–8. DOI: 10.1182/blood-2002-02-0420.Murphy K.M., Levis M., Hafez M.J., et al. Detection of FLT3 internal tandem duplication and D835 mutations by a multiplex polymerase chain reaction and capillary electrophoresis assay. J Mol Diagn. 2003;5(2):96–102. DOI: 10.1016/S1525-1578(10)60458-8.Murphy K.M., Levis M., Hafez M.J., et al. Detection of FLT3 internal tandem duplication and D835 mutations by a multiplex polymerase chain reaction and capillary electrophoresis assay. J Mol Diagn. 2003;5(2):96–102. DOI: 10.1016/S1525-1578(10)60458-8.Диагностика заболеваний системы крови. Практическое руководство. Под ред. Е.Н. Паровичниковой, И.В. Гальцевой. М.: Практика; 2024. 560 с.Diagnostics of blood system diseases. Practical guide. Edited by. Parovichnikova E.N., Galtseva I.V. ed. Moscow: Praktika, 2024:560 (In Russian).Кашлакова А.И., Паровичникова Е.Н., Бидерман Б.В. и др. Определение молекулярно-генетического профиля у взрослых больных острыми миелоидными лейкозами методом секвенирования нового поколения. Гематология и трансфузиология. 2020;65(4):444–59. DOI: 10.35754/0234-5730-2020-65-4-444-459.Kashlakova A.I., Parovichnikova E.N., Biderman B.V., et al. Next-generation sequencing-based molecular genetic profi ling in adults with acute myeloid leukaemia. Gematologiya i transfuziologiya 2020;65(4):444–59 (In Russian). DOI: 10.35754/0234-5730-2020-65-4-444-459.Sakaguchi M., Nakajima N., Yamaguchi H., et al. The sensitivity of the FLT3- ITD detection method is an important consideration when diagnosing acute myeloid leukemia. Leuk Res Rep. 2020;13:100198. DOI: 10.1016/j.lrr.2020.100198.Sakaguchi M., Nakajima N., Yamaguchi H, et al. The sensitivity of the FLT3-ITD detection method is an important consideration when diagnosing acute myeloid leukemia. Leuk Res Rep. 2020;13:100198. DOI: 10.1016/j.lrr.2020.100198.Beierl K., Tseng L.H., Beierl R., et al. Detection of minor clones with internal tandem duplication mutations of FLT3 gene in acute myeloid leukemia using delta-PCR. Diagn Mol Pathol. 2013;22(1):1–9. DOI: 10.1097/PDM.0b013e31825d81f4.Beierl K., Tseng L.H., Beierl R., et al. Detection of minor clones with internal tandem duplication mutations of FLT3 gene in acute myeloid leukemia using delta-PCR. Diagn Mol Pathol. 2013;22(1):1–9. DOI: 10.1097/PDM.0b013e31825d81f4.Lin M.T., Tseng L.H., Beierl K., et al. Tandem duplication PCR: an ultrasensitive assay for the detection of internal tandem duplications of the FLT3 gene. Diagn Mol Pathol. 2013;22(3):149–55. DOI: 10.1097/PDM.0b013e31828308a1.Lin M.T., Tseng L.H., Beierl K., et al. Tandem duplication PCR: an ultrasensitive assay for the detection of internal tandem duplications of the FLT3 gene. Diagn Mol Pathol. 2013;22(3):149–55. DOI: 10.1097/PDM.0b013e31828308a1.Loo S., Dillon R., Ivey A., et al. Pretransplant FLT3-ITD MRD assessed by high-sensitivity PCR-NGS determines posttransplant clinical outcome. Blood. 2022;140:2407–11. DOI: 10.1182/blood.2022016567.Loo S., Dillon R., Ivey A., et al. Pretransplant FLT3-ITD MRD assessed by high-sensitivity PCR-NGS determines posttransplant clinical outcome. Blood. 2022;140:2407–11. DOI: 10.1182/blood.2022016567.Grob T., Sanders M.A., Vonk C.M., et al. Prognostic Value of FLT3-Internal Tandem Duplication Residual Disease in Acute Myeloid Leukemia. J. Clin. Oncol. 2023;41:756–65. DOI: 10.1200/JCO.22.00715.Grob T., Sanders M.A., Vonk C.M., et al. Prognostic Value of FLT3-Internal Tandem Duplication Residual Disease in Acute Myeloid Leukemia. J. Clin. Oncol. 2023;41:756–65. DOI: 10.1200/JCO.22.00715.Lin M.T., Tseng L.H., Dudley J.C., et al. A Novel Tandem Duplication Assay to Detect Minimal Residual Disease in FLT3/ITD AML. Mol Diagn Ther. 2015;19(6):409–17. DOI: 10.1007/s40291-015-0170-3.Lin M.T., Tseng L.H., Dudley J.C., et al. A Novel Tandem Duplication Assay to Detect Minimal Residual Disease in FLT3/ITD AML. Mol Diagn Ther. 2015;19(6):409–17. DOI: 10.1007/s40291-015-0170-3.Blätte T.J., Schmalbrock L.K., Skambraks S., et al. getITD for FLT3-ITD-based MRD monitoring in AML. Leukemia. 2019;33(10):2535–9. DOI: 10.1038/s41375-019-0483-z.Blätte T.J., Schmalbrock L.K., Skambraks S., et al. getITD for FLT3-ITD-based MRD monitoring in AML. Leukemia. 2019;33(10):2535–9. DOI: 10.1038/s41375-019-0483-z.Bibault J.E., Figeac M., Hélevaut N., et al. Next-generation sequencing of FLT3 internal tandem duplications for minimal residual disease monitoring in acute myeloid leukemia. Oncotarget. 2015;6(26):22812–21. DOI: 10.18632/oncotarget.4333.Bibault J.E., Figeac M., Hélevaut N., et al. Next-generation sequencing of FLT3 internal tandem duplications for minimal residual disease monitoring in acute myeloid leukemia. Oncotarget. 2015;6(26):22812–21. DOI: 10.18632/oncotarget.4333.Falini B., Nicoletti I., Martelli M.F., Mecucci C. Acute myeloid leukemia carrying cytoplasmic/mutated nucleophosmin (NPMc+ AML): Biologic and clinical features. Blood. 2007;109:874–85. DOI: 10.1182/blood-2006-07-012252.Falini B., Nicoletti I., Martelli M.F., Mecucci C. Acute myeloid leukemia carrying cytoplasmic/mutated nucleophosmin (NPMc+ AML): Biologic and clinical features. Blood. 2007;109:874–85. DOI: 10.1182/blood-2006-07-012252.Heath E.M., Chan S.M., Minden M.D., et al. Biological and clinical consequences of NPM1 mutations in AML. Leukemia. 2017;31:798–807. DOI: 10.1038/leu.2017.30.Heath E.M., Chan S.M., Minden M.D., Murphy T., Shlush L.I., Schimmer A.D. Biological and clinical consequences of NPM1 mutations in AML. Leukemia. 2017;31:798–807. DOI: 10.1038/leu.2017.30.Kihara R, Nagata Y, Kiyoi H., et al. Comprehensive analysis of genetic alterations and their prognostic impacts in adult acute myeloid leukemia patients. Leukemia. 2014;28(8):1586–95. DOI: 10.1038/leu.2014.55.Kihara R, Nagata Y, Kiyoi H., et al. Comprehensive analysis of genetic alterations and their prognostic impacts in adult acute myeloid leukemia patients. Leukemia. 2014;28(8):1586–95. DOI: 10.1038/leu.2014.55.Döhner K., Schlenk R.F., Habdank M., et al. Mutant nucleophosmin (NPM1) predicts favorable prognosis in younger adults with acute myeloid leukemia and normal cytogenetics: Interaction with other gene mutations. Blood. 2005;106:3740–6. DOI: 10.1182/blood-2005-05-2164.Döhner K., Schlenk R.F., Habdank M., et al. Mutant nucleophosmin (NPM1) predicts favorable prognosis in younger adults with acute myeloid leukemia and normal cytogenetics: Interaction with other gene mutations. Blood. 2005;106:3740–6. DOI: 10.1182/blood-2005-05-2164.Jahn N., Jahn E., Saadati M., et al. Genomic landscape of patients with FLT3- mutated acute myeloid leukemia (AML) treated within the CALGB 10603/RATIFY trial. Leukemia. 2022;36(9):2218–27. DOI: 10.1038/s41375-022-01650-w.Jahn N., Jahn E., Saadati M., et al. Genomic landscape of patients with FLT3- mutated acute myeloid leukemia (AML) treated within the CALGB 10603/RATIFY trial. Leukemia. 2022;36(9):2218–27. DOI: 10.1038/s41375-022-01650-w.Severens J.F., Karakaslar E.O., van der Reijden, B.A., et al. Mapping AML heterogeneity — multi-cohort transcriptomic analysis identifies novel clusters and divergent ex-vivo drug responses. Leukemia. 2024;38:751–61. DOI: 10.1038/s41375-024-02137-6.Severens, J.F., Karakaslar, E.O., van der Reijden, B.A. et al. Mapping AML heterogeneity — multi-cohort transcriptomic analysis identifi es novel clusters and divergent ex-vivo drug responses. Leukemia. 2024;38:751–61. DOI: 10.1038/s41375-024-02137-6.Tyner J.W., Tognon C.E., Bottomly D., et al. Functional genomic landscape of acute myeloid leukaemia. Nature. 2018;562:526–53. DOI: 10.1038/s41586-018-0623-z.Tyner, J.W., Tognon, C.E., Bottomly, D. et al. Functional genomic landscape of acute myeloid leukaemia. Nature. 2018;562:526–53. DOI: 10.1038/s41586-018-0623-z.Yu J., Sun J., Du Y., Chang C. Genomic Landscape of Acute Myeloid Leukemia (AML) on the Basis of 2017 ELN Classification and Other Mutations in Adult AML — Single Healthcare System Data. Blood. 2020;136(Suppl. 1):3–4. DOI: 10.1182/blood-2020-139976.Yu J., Sun J., Du Y., Chang C. Genomic Landscape of Acute Myeloid Leukemia (AML) on the Basis of 2017 ELN Classifi cation and Other Mutations in Adult AML — Single Healthcare System Data. Blood. 2020;136(Suppl. 1):3–4. DOI: 10.1182/blood-2020-139976.Пехова К.А., Сидорова Ю.В., Северина Н.А. и др. Генетический ландшафт острых миелоидных лейкозов, протекающих с лейкоцитозом. Онкогематология. 2023;18(3):102–14. DOI: 10.17650/1818-8346-2023-18-3-102-114.Pekhova K.A., Sidorova Yu.V., Severina N.A., et al. Genetic landscape of acute myeloid leukemias with leukocytosis. Oncohematologiya. 2023;18(3):102– 14 (In Russian). DOI 10.17650/1818-8346-2023-18-3-102-114.Swerdlow S.H., Campo E., Harris N.L., et al. WHO classification of tumors of haematopoietic and lymphoid tissues. Lyon: IARC, 2017:130–55.Swerdlow S.H., Campo E., Harris N.L., et al. WHO classifi cation of tumors of haematopoietic and lymphoid tissues. Lyon: IARC, 2017:130–55.Krönke J., Schlenk R.F., Jensen K-O., et al. Monitoring of minimal residual disease in NPM1-mutated acute myeloid leukemia: a study from the German-Austrian acute myeloid leukemia study group. J Clin Oncol. 2011;29:2709–16. DOI: 10.1200/JCO.2011.35.0371.Krönke J., Schlenk R.F., Jensen K-O., et al. Monitoring of minimal residual disease in NPM1-mutated acute myeloid leukemia: a study from the German-Austrian acute myeloid leukemia study group. J Clin Oncol. 2011;29:2709–16. DOI: 10.1200/JCO.2011.35.0371.Ivey A., Hills R.K., Simpson M.A., et al. Assessment of minimal residual disease in standard-risk AML. N. Engl. J. Med. 2016;374:422–33. DOI: 10.1056/NEJMoa1507471.Ivey A., Hills R.K., Simpson M.A., et al. Assessment of minimal residual disease in standard-risk AML. N. Engl. J. Med. 2016;374:422–33. DOI: 10.1056/NEJMoa1507471.Schuurhuis G.J., Heuser M., Freeman S., et al. Minimal/measurable residual disease in AML: A consensus document from the European LeukemiaNet MRD Working Party. Blood. 2018;131:1275–91. DOI: 10.1182/blood-2017-09-801498.Schuurhuis G.J., Heuser M., Freeman S., et al. Minimal/measurable residual disease in AML: A consensus document from the European LeukemiaNet MRD Working Party. Blood. 2018;131:1275–91. DOI: 10.1182/blood-2017-09-801498.Heuser M., Freeman S.D., Ossenkoppele G.J., et al. 2021 Update on MRD in acute myeloid leukemia: a consensus document from the European LeukemiaNet MRD Working Party. Blood. 2021;138(26):2753–67. DOI: 10.1182/blood.2021013626.Heuser M, Freeman SD, Ossenkoppele GJ, et al. 2021 Update on MRD in acute myeloid leukemia: a consensus document from the European LeukemiaNet MRD Working Party. Blood. 2021;138(26):2753–67. DOI: 10.1182/blood.2021013626.Dohner H., Estey E., Grimwade D., et al. Diagnosis and management of AML in adults: 2017 ELN recommendations from an international expert panel. Blood. 2017;129:424–47. DOI: 10.1182/blood-2016-08-733196.Dohner H., Estey E., Grimwade D., et al. Diagnosis and management of AML in adults: 2017 ELN recommendations from an international expert panel. Blood. 2017;129:424–47. DOI: 10.1182/blood-2016-08-733196.Алгоритмы диагностики и протоколы лечения заболеваний системы крови. Под ред. Паровичниковой Е.Н. М.: Практика; 2024:621–81.Diagnostic algorithms and protocols for the treatment of diseases of the blood system. Ed. Parovichnikova E.N. Moscow: Praktika; 2024:621–81 (In Russian).Сидорова Ю.В., Сорокина Т.В., Бидерман Б.В. и др. Определение минимальной остаточной болезни у больных В-клеточным хроническим лимфолейкозом методом пациент-специфичной ПЦР. Клиническая лабораторная диагностика. 2011; (12): 22-35.Sidorova Yu.V., Sorokina T.V., Biderman B.V., et al. The detection of minimal residual disease in patients with chronic B-cell lymphatic leukemia using patientspecifi ed polymerase chain reaction. Klinicheskaya I Laboratornaya Diagnostika. 2011;12:22–35 (In Russian). DOI: 10.35754/0234-5730-2025-70-1-8-26.Сидорова Ю.В., Северина Н.А., Бидерман Б.В. и др. Определение мутаций FLT3 и мониторинг МОБ минимальной остаточной болезни при FLT3- позитивном остром миелоидном лейкозе. Гематология и трансфузиология. 2025:70(1):8–26. DOI: 10.35754/0234-5730-2025-70-1-8-26.Sidorova Yu.V. 1, Severina N.A. 1, Biderman B.V., et al. Determination of FLT3 mutations and MRD minimal residual deasease monitoring in FLT3-positive acute myeloid leukemiaAML. Gematologiâ i transfuziologiâ. 2025:70(1):8–26 (In Russian). DOI: 10.35754/0234-5730-2025-70-1-8-26.Сидорова Ю.В., Северина Н.А., Судариков А.Б. Тест-система и способ выявления A, B, D мутаций гена NPM1 для количественного определения минимальной остаточной болезни. Изобретения и полезные модели. 2024. Бюл. 33. Патент № 2830545.Sidorova Yu.V., Severina N.A., Sudarikov A.B. Test system and method for detecting A, B, D mutations of NPM1 gene for quantitative determination of minimal residual disease. Izobreteniya i polezniye modeli. Bull. 33. Patent № 2830545 (In Russian).Schranz K., Hubmann M., Harin E., et al. Clonal heterogeneity of FLT3- ITD detected by high-throughput amplicon sequencing correlates with adverse prognosis in acute myeloid leukemia. Oncotarget. 2018;9(53):30128–45. DOI: 10.18632/oncotarget.25729.Schranz K., Hubmann M., Harin E., et al. Clonal heterogeneity of FLT3- ITD detected by high-throughput amplicon sequencing correlates with adverse prognosis in acute myeloid leukemia. Oncotarget. 2018;9(53):30128–45. DOI: 10.18632/oncotarget.25729.Zuffa E., Franchini E., Papayannidis C., et al. Revealing very small FLT3 ITD mutated clones by ultra-deep sequencing analysis has important clinical implications in AML patients. Oncotarget. 2015;6(31):31284–94. DOI: 10.18632/oncotarget.5161.Zuffa E., Franchini E., Papayannidis C., et al. Revealing very small FLT3 ITD mutated clones by ultra-deep sequencing analysis has important clinical implications in AML patients. Oncotarget. 2015;6(31):31284–94. DOI: 10.18632/oncotarget.5161.Ottone T., Zaza S., Divona M., et al. Identification of emerging FLT3 ITD-positive clones during clinical remission and kinetics of disease relapse in acute myeloid leukaemia with mutated nucleophosmin. Br J Haematol. 2013;161(4):533– 40. DOI: 10.1111/bjh.12288.Ottone T., Zaza S., Divona M., et al. Identifi cation of emerging FLT3 ITD-positive clones during clinical remission and kinetics of disease relapse in acute myeloid leukaemia with mutated nucleophosmin. Br J Haematol. 2013;161(4):533– 40. DOI: 10.1111/bjh.12288.Sakaguchi M., Yamaguchi H., Najima Y., et al. Prognostic impact of low allelic ratio FLT3-ITD and NPM1 mutation in acute myeloid leukemia. Blood Adv. 2018;2(20):2744–54. DOI: 10.1182/bloodadvances.2018020305.Sakaguchi M., Yamaguchi H., Najima Y., et al. Prognostic impact of low allelic ratio FLT3-ITD and NPM1 mutation in acute myeloid leukemia. Blood Adv. 2018;2(20):2744–54. DOI: 10.1182/bloodadvances.2018020305.Thiede C., Steudel C., Mohr B., et al. Analysis of FLT3-activating mutations in 979 patients with acute myelogenous leukemia: association with FAB subtypes and identification of subgroups with poor prognosis. Blood. 2002;99(12):4326–35. DOI: 10.1182/blood.v99.12.4326.Thiede C, Steudel C, Mohr B, et al. Analysis of FLT3-activating mutations in 979 patients with acute myelogenous leukemia: association with FAB subtypes and identifi cation of subgroups with poor prognosis. Blood 2002;99(12):4326– 35. DOI: 10.1182/blood.v99.12.4326.Chen F., Sun J., Yin, C., et al. Impact of FLT3-ITD allele ratio and ITD length on therapeutic outcome in cytogenetically normal AML patients without NPM1 mutation. Bone Marrow Transplant. 2020;55:740–8. DOI: 10.1038/s41409-019-0721-z.Chen F., Sun J., Yin, C., et al. Impact of FLT3-ITD allele ratio and ITD length on therapeutic outcome in cytogenetically normal AML patients without NPM1 mutation. Bone Marrow Transplant. 2020;55:740–8. DOI: 10.1038/s41409-019-0721-z.Schlenk R.F., Kayser S., Bullinger L., et al. German-Austrian AML Study Group. Differential impact of allelic ratio and insertion site in FLT3-ITD-positive AML with respect to allogeneic transplantation. Blood. 2014;124(23):3441–9. DOI: 10.1182/blood-2014-05-578070.Schlenk R.F., Kayser S., Bullinger L., et al. German-Austrian AML Study Group. Differential impact of allelic ratio and insertion site in FLT3-ITD-positive AML with respect to allogeneic transplantation. Blood. 2014;124(23):3441–9. DOI: 10.1182/blood-2014-05-578070.Abou Dalle I, Ghorab A, Patel K, et al. Impact of numerical variation, allele burden, mutation length and co-occurring mutations on the efficacy of tyrosine kinase inhibitors in newly diagnosed FLT3- mutant acute myeloid leukemia. Blood Cancer J. 2020;10(5):48. DOI: 10.1038/s41408-020-0318-148.Abou Dalle I, Ghorab A, Patel K, et al. Impact of numerical variation, allele burden, mutation length and co-occurring mutations on the effi cacy of tyrosine kinase inhibitors in newly diagnosed FLT3- mutant acute myeloid leukemia. Blood Cancer J. 2020;10(5):48. DOI: 10.1038/s41408-020-0318-1 48.Döhner H., Wei A.H., Appelbaum F.R., et al. Diagnosis and management of AML in adults: 2022 recommendations from an international expert panel on behalf of the ELN. Blood. 2022;140:1345–77. DOI: 10.1182/blood.2022016867Döhner H., Wei A.H., Appelbaum F.R., et al. Diagnosis and management of AML in adults: 2022 recommendations from an international expert panel on behalf of the ELN. Blood. 2022;140:1345–77. DOI: 10.1182/blood.2022016867.Larson R.A., Mandrekar S.J., Huebner L.J., et al. Midostaurin reduces relapse in FLT3-mutant acute myeloid leukemia: the Alliance CALGB 10603/RATIFY trial. Leukemia. 2021;35(9):2539–51. DOI: 10.1038/s41375-021-01179-4.Larson R.A., Mandrekar S.J., Huebner L.J., et al. Midostaurin reduces relapse in FLT3-mutant acute myeloid leukemia: the Alliance CALGB 10603/RATIFY trial. Leukemia. 2021;35(9):2539–51. DOI: 10.1038/s41375-021-01179-4.Döhner H., Weber D., Krzykalla J., et al. Midostaurin plus intensive chemotherapy for younger and older patients with AML and FLT3 internal tandem duplications. Blood Adv. 2022;6:5345–55. DOI: 10.1182/bloodadvances.2022007223.Döhner H., Weber D., Krzykalla J., et al. Midostaurin plus intensive chemotherapy for younger and older patients with AML and FLT3 internal tandem duplications. Blood Adv. 2022;6:5345–55. DOI: 10.1182/bloodadvances.2022007223.Клинические рекомендации № 131 «Острые миелоидные лейкозы. Взрослые». 2024. URL: https://cr.minzdrav.gov.ru/view-cr/131_2Clinical recomendations No. 131. “Acute myeloid leukemia. Adult” (In Russian). 2024. URL: https://cr.minzdrav.gov.ru/view-cr/131_2Pollyea D.A., Bixby D., Perl A., et al. NCCN Guidelines Insights: Acute Myeloid Leukemia, Version 2.2021. J Natl Compr Canc Netw. 2021;19(1):16–27. DOI: 10.6004/jnccn.2021.0002.Pollyea D.A., Bixby D., Perl A., et al. NCCN Guidelines Insights: Acute Myeloid Leukemia, Version 2.2021. J Natl Compr Canc Netw. 2021;19(1):16–27. DOI: 10.6004/jnccn.2021.0002.

The authors declare that there are no conflicts of interest present.