Новейший метод секвенирование нового поколения (Next Generation Sequencing, NGS) позволяет выявлять генетическое и эпигенетическое разнообразие хронического лимфолейкоза (ХЛЛ), декодируя мутационный статус и оценивая его влияние на течение заболевания. Учитывая, что тактика лечения пациентов с ХЛЛ претерпела революционные изменения в последние два десятилетия, детекция драйверных мутаций позволит выбирать оптимальные терапевтические схемы и содействовать разработке и дальнейшему внедрению новейших таргетных препаратов и их комбинаций. Риск-адаптированный подход к терапии ХЛЛ, основанный на группах генетического риска, будет являться базисом персонифицированного лечения. Целью настоящей работы является оценка мутационного статуса пациентов сХЛЛ с использованием разработанной лимфоидной таргетной NGS-панели и изучение возможной корреляции мутационного статуса с клиническими характеристиками заболевания. В проспективное исследование было включено 68 пациентов с ХЛЛ: нелеченые (n=56) и рецидивирующие и рефрактерные формы (n=12). С целью апробации «Лимфоидной таргетной NGS панели генов» было выполнено исследование на образцах ДНК пациентов (n=19) с ХЛЛ. Использование NGS позволило выявить генетические аберрации у всех исследуемых пациентов. Общее количество мутаций составило 767. Полученные нами данные пилотного исследования демонстрируют возможность применения технологии NGS в клинической практике. Для оценки корреляции клинико-лабораторных параметров с мутационным статусом пациентов сХЛЛ исследование будет продолжено. Учитывая, что информация о прогностической значимости ряда выявленных мутаций в настоящее время отсутствует, требуются дополнительные исследования с целью определения их влияния на течение ХЛЛ. The newest method - Next Generation Sequencing (NGS) - allows detecting the genetic and epigenetic diversity of chronic lymphocytic leukemia (CLL), decoding the mutational status and assessing its effect on the course of the disease. Considering that the tactics of treatment of patients with CLL have undergone revolutionary changes in the past two decades, the detection of driver mutations will allow choosing the optimal therapeutic regimens and promoting the development and further implementation of the latest targeted drugs and their combinations. A risk-adapted approach to CLL therapy based on genetic risk groups will be the base for personalized treatment. This work aims to assess the mutational status of patients with CLL using the developed “Lymphoid targeted NGS panel” and to study the possible correlation of the mutational status with the clinical characteristics of the disease. The prospective study included 68 patients with CLL: untreated (n=56) and relapse/refractory (n=12). In order to test the “Lymphoid targeted NGS gene panel”, the study was carried out on DNA samples from patients (n=19) with CLL. The use of NGS let to identify genetic aberrations in all patients. The total number of mutations was 767. Our pilot study data demonstrate the possibility of using NGS technology in clinical practice. The study will be continued to assess the correlation of clinical and laboratory parameters with CLL’s mutational status. Considering that there is currently no information on the prognostic significance of several identified mutations, additional studies are required.

Background:According to various authors, the frequency of concurrent myelodysplastic syndrome (MDS) in patients with chronic lymphocytic leukemia (CLL) is 5%. The observation of patients with combination of lympho‐ and myeloproliferative diseases is clinical interest, due to the rare occurrence, poorer outcomes and absence of optimal treatment strategy for this category of patients.Aims:We present a case of a 71‐year‐old woman with CLL and acute myeloid leukemia (AML) developed from MDS.Methods:The baseline diagnosis CLL III stage (Rai) was established in 2016 at 68 years. FISH: trisomy12 – 9%; del13 – 20.5%; del17p – 17.5%. CT scan demonstrated lymphadenopathy (axillary, epitrochlear, mediastinal, paraaortic). The first line therapy: Ibrutinib 420 mg daily, and Rituximab 375 mg/m2 D1 C1 and 500 mg/m2 D1 C2‐6 of each 28‐day cycle. Despite the treatment, patient still had moderate/severe anemia without reduction of transfusion dependency. The disease evaluation (DE) after cycle 6 of therapy showed the presence of 13.4% blasts and dysplasia in erythroid, granulocyte and megakaryocyte lines (≥10%) in BM. Flow cytometry (FCM) of BM demonstrated the population of myeloid blasts. The BM's biopsy showed combined lesions: MDS with excess of blasts‐2 and CLL (5‐7% of infiltrate). The lymph nodes, spleen and liver were normal sizes by CT scan. The response of CLL was evaluated as partial remission (PR) and the second diagnosis was established: MDS with excess of blasts‐2 (IPSS: high risk, IPSS‐R: very high risk). Azacytidine 75 mg/m2 D1‐7 of each 28‐day cycle, SC was started. Simultaneous, therapy with ibrutinib 420 mg daily was ongoing. After 4 cycles of treatment the patient became transfusion independent; HB level stabilized at 110–115 g/L; BM blasts decreased to 9.8%, and dysplasia changes persisted only in granulocyte line. We observed the decrease of minimal residual disease (MRD) of CLL: 8.6%→5.4%→3.8%. The toxicity grade 3‐4 during therapy was not observed. However, after 8 cycle of therapy the results of laboratory tests showed transformation to AML: HB–73 g/L, WBC–1.6x109/L (NEUT–27%, BANDS–1%, LYM–66%, MON–6%), PLT–44x109/L; BM blasts–24%; FCM of BM ‐ population of myeloid blasts; normal karyotype 46, XX; mutations in genes FLT3, NMP1, BCR/ABL, CBFb/MYH1, RUNX1, TP53, IDH1/2, c‐KIT, ASXL1 were not detected. Treatment with ibrutinib was stopped. Cytozar 38 mg D1‐10 of each 28‐day cycle, SC, and venetoclax 600 mg daily were started.Results:After 1stcycle of therapy: complete remission (CR) with incomplete BM recovery. After 2ndcycle – complete BM recovery. By FCM MRD negative status (MRDneg) AML and MRD positive status (MRDpos) CLL were detected. Patient is ongoing to intake cytozar and venetoclax. Presently, the MRDneg CR of AML and MRDneg PR of CLL are lasting.Summary/Conclusion:Our findings suggest that use of targeted drug in the treatment of patients with AML and CLL is possible and very effective. Also, treatment combination of low dose cytozar with venetoclax is powerful and shows clinical significance. Due to the small number of patients with this combined pathology, further studies are needed to confirm the results obtained.

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.