scholarly journals A Novel System for Semiautomatic Sample Processing in Chronic Myeloid Leukaemia: Increasing Throughput without Impacting on Molecular Monitoring at Time of SARS-CoV-2 Pandemic

Diagnostics ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 1502
Author(s):  
Stefania Stella ◽  
Silvia Rita Vitale ◽  
Michele Massimino ◽  
Adriana Puma ◽  
Cristina Tomarchio ◽  
...  
Keyword(s):  
Chronic Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Kinase Inhibitors ◽  
Quantitative Polymerase Chain Reaction ◽  
White Blood Cells ◽  
Rna Isolation ◽  
Molecular Testing ◽  
Molecular Response ◽  
Molecular Monitoring ◽  
Semiautomatic System

Molecular testing of the BCR-ABL1 transcript via real-time quantitative-polymerase-chain-reaction is the most sensitive approach for monitoring the response to tyrosine-kinase-inhibitors therapy in chronic myeloid leukaemia (CML) patients. Each stage of the molecular procedure has been standardized and optimized, including the total white blood cells (WBCs) and RNA isolation methods. Here, we compare the performance of our current manual protocol to a newly semiautomatic method based on the Biomek i-5 Automated Workstations integrated with the CytoFLEX Flow Cytometer, followed by the automatic QIAsymphony system to facilitate high-throughput processing samples and reduce the hands-on time and the risk associated with SARS-CoV-2. The recovery efficiency was investigated in blood samples from 100 adults with CML. We observe a 100% of concordance between the two methods, with similar total WBCs isolated (median 1.137 × 106 for manual method vs. 1.076 × 106 for semiautomatic system) and a comparable quality and quantity of RNA extracted (median 103 ng/μL with manual isolation kit vs. 99.95 ng/μL with the QIAsymphony system). Moreover, by stratifying patients according to their BCR-ABL1 transcript levels, we obtained similar BCR-ABL1/ABL1IS values and ABL1 copies, and matched samples were assigned to the same group of molecular response. We conclude that this newly semiautomatic workflow has a performance comparable to our more laborious standard manual, which can be replaced, particularly when specimens from patients with suspected or confirmed SARS-CoV-2 infection need to be processed.

scholarly journals Treatment-Free Remission in Chronic Myeloid Leukaemia: Can We Identify Prognostic Factors?

2021 ◽  
Author(s):  
Hilbeen Hisham Saifullah ◽  
Claire Marie Lucas
Keyword(s):  
Prognostic Factors ◽  
Chronic Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Kinase Inhibitors ◽  
Chronic Phase ◽  
Molecular Response ◽  
Predicting Success ◽  
Treatment Free Remission ◽  
Almost All ◽  
Tki Discontinuation

Following the development of tyrosine kinase inhibitors (TKI), the survival of patients with chronic myeloid leukaemia (CML) drastically improved. With the introduction of these agents, CML is now considered a chronic disease, for some patients. Taking into consideration the side effects, toxicity, and high cost, discontinuing TKIs became a goal for patients with chronic phase CML. Patients who achieved deep molecular response (DMR) and discontinued TKI, remained in treatment-free remission (TFR). Currently, the data from the published literature demonstrate that 40-60% of patients achieve TFR, with relapses occurring within the first six months. In addition, almost all patients who relapsed regained a molecular response upon re-treatment, indicating TKI discontinuation is safe. However, there is still a gap in the understanding the mechanisms behind TFR, and whether there are prognostic factors that can predict the best candidates who qualify for TKI discontinuation with a view to keeping them in TFR. Furthermore, the information about a second TFR attempt and the role of gradual de-escalation of TKI before complete cessation is limited. This review highlights the factors predicting success or failure of TFR. In addition, it ex-amines the feasibility of a second TFR attempt after the failure of the first one, and the current guidelines concerning TFR in clinical practice.

scholarly journals Biomarkers for BCR-ABL1 as a Potential Therapeutic Option Among Adult Chronic Myeloid Leukaemia Treated With Tyrosine Kinase Inhibitors at Molecular Response in Multi-racial Malaysia: Hsa-mir-181a-5p for Decrement; hsa-mir-182-5p and hsa-mir-26a-5p for Increment.

2020 ◽  
Author(s):  
Aliza Mohd Yacob ◽  
Nor Asiah Muhammad ◽  
Kian Meng Chang ◽  
Hamidah Akmal Hisham ◽  
Yuslina Mat Yusoff ◽  
...  
Keyword(s):  
Chronic Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Kinase Inhibitors ◽  
Therapeutic Option ◽  
P Value ◽  
Molecular Response ◽  
Down Regulation ◽  
Molecular Responses ◽  
Number Of Patients ◽  
Long Term Treatment

Abstract BackgroundTyrosine kinase inhibitors (TKIs) as first-line therapy for Chronic Myeloid Leukaemia (CML) show a high success rate. However, low number of patients with long-term treatment-free remission (TFR) is observed. Molecular relapse after imatinib discontinuation occurred at 50% at 24 months with 80% occurrence within the first 6 months. One of the reasons for relapse is untimely TKIs discontinuation caused by large errors from estimates at very low-level or undetectable disease, thus warranted new biomarkers for CML. MethodsNext Generation Sequencing (NGS) was used to identify microRNAs (miRNAs) at molecular response in CML adult patients receiving TKIs treatment. A total of 86 samples were collected, 30 from CML patients responsive and 28 from non-responsive to imatinib therapy, and 28 from blood donors. NGS was conducted whereby 18 miRNAs were selected and validated by real time RT-qPCR in triplicate. ResultsHsa-miR-181a-5p was expressed significantly (p-value<0.05) with 2.14 and 2.3 fold down-regulation in both patient groups respectively while hsa-miR-182-5p and hsa-miR-26a-5p were significant only in the non-responsive group with 2.08 and 2.39 fold up-regulation respectively. The down-regulation was consistent with decreased amount of BCR-ABL1 in patients taking TKIs regardless of molecular responses. The up-regulation was consistent with substantial presence of BCR-ABL1 in CML patients treated with TKIs at molecular response. ConclusionTherefore, these miRNAs have potential as new therapeutic biomarkers for BCR-ABL1 status in adult CML patients treated with TKIs at molecular responses. These could improve current approaches and require further analysis to look for targets of these miRNAs in CML.

scholarly journals Expert Opinion on the Treatment of Refractory Chronic Phase Chronic Myeloid Leukaemia

2017 ◽  
Vol 13 (01) ◽  
pp. 17
Author(s):  
Tim Hughes ◽  
Giuseppe Saglio ◽  
◽  
Keyword(s):  
Chronic Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Second Generation ◽  
Kinase Inhibitors ◽  
Chronic Phase ◽  
Molecular Response ◽  
Line Treatment ◽  
T315i Mutation ◽  
Third Line ◽  
Third Line Treatment

The development and clinical availability of second-generation tyrosine kinase inhibitors (TKIs) for the treatment of patients who discontinue imatinib therapy has further improved the outlook for patients with chronic phase chronic myeloid leukaemia (CP-CML). There is, however, uncertainty surrounding how best to treat patients after failing second-generation TKIs. A three-section questionnaire was devised by chronic myeloid leukaemia experts to address questions surrounding this issue. Responses were received from 14 out of 34 experts (41.2%). Generally, a reasonable consensus was found among the responses for most issues. There was a complete consensus that ponatinib was suitable for all patients carrying the T315I mutation regardless of the molecular response to prior treatment. There was also complete consensus that allografting is appropriate in any patient who has had blast crises and is back in a second chronic phase. More recommendations for third-line treatment of CP-CML patients are necessary.

scholarly journals Molecular Testing in CML between Old and New Methods: Are We at a Turning Point?

2020 ◽  
Vol 9 (12) ◽  
pp. 3865
Author(s):  
Simona Soverini ◽  
Simona Bernardi ◽  
Sara Galimberti
Keyword(s):  
Kinase Inhibitor ◽  
Residual Disease ◽  
Quantitative Polymerase Chain Reaction ◽  
Mutation Testing ◽  
Response Monitoring ◽  
Molecular Testing ◽  
Molecular Response ◽  
Molecular Monitoring ◽  
Clinical Value ◽  
Novel Technologies

Molecular monitoring of minimal residual disease (MRD) and BCR-ABL1 kinase domain (KD) mutation testing have a well consolidated role in the routine management of chronic myeloid leukemia (CML) patients, as they provide precious information for therapeutic decision-making. Molecular response levels are used to define whether a patient has an “optimal”, “warning”, or “failure” response to tyrosine kinase inhibitor (TKI) therapy. Mutation status may be useful to decide whether TKI therapy should be changed and which alternative TKI (or TKIs) are most likely to be effective. Real-time quantitative polymerase chain reaction (RQ-qPCR) and Sanger sequencing are currently the gold standard for molecular response monitoring and mutation testing, respectively. However, in recent years, novel technologies such as digital PCR (dPCR) and next-generation sequencing (NGS) have been evaluated. Here, we critically describe the main features of these old and novel technologies, provide an overview of the recently published studies assessing the potential clinical value of dPCR and NGS, and discuss how the state of the art might evolve in the next years.

scholarly journals Molecular monitoring in CML: how deep? How often? How should it influence therapy?

Blood ◽  
2018 ◽  
Vol 132 (20) ◽  
pp. 2125-2133 ◽  
Author(s):  
Naranie Shanmuganathan ◽  
Timothy P. Hughes
Keyword(s):  
Messenger Rna ◽  
Kinase Inhibitors ◽  
Quantitative Polymerase Chain Reaction ◽  
Response Assessment ◽  
Molecular Response ◽  
Molecular Monitoring ◽  
Optimal Outcomes ◽  
Patients At Risk ◽  
Failure Depth ◽  
Treatment Free Remission

Abstract With the advent of tyrosine kinase inhibitors (TKIs), the goals of therapy in chronic myeloid leukemia (CML) are steadily shifting. Long-term disease control on TKI therapy has been the goal and expectation for most patients. More recently, treatment-free remission (TFR) has entered mainstream practice and is increasingly being adopted as the main goal of therapy. This therapeutic shift not only influences TKI selection but also, has necessitated the refinement and dissemination of highly sensitive and accurate molecular monitoring techniques. Measurement of BCR-ABL1 messenger RNA expression through reverse transcription quantitative polymerase chain reaction, reported according to the International Scale, has become the primary tool for response assessment in CML. Achieving specific time-dependent molecular milestones, as defined by global therapeutic guidelines, has been established as critical in maximizing optimal outcomes while identifying patients at risk of therapy failure. Depth and duration of a deep molecular response have become the new therapeutic targets in patients considered for TFR. Consequently, molecular monitoring in CML has become even more critical to ongoing response assessment, identifying patients with TKI resistance and poor drug adherence, and enabling TFR to be attempted safely and effectively.

scholarly journals Comparison of Response to Treatment with Imatinib Versus Nilotinib in the Initial Three Months in Chronic Myeloid Leukaemia

2021 ◽  
Vol 8 (24) ◽  
pp. 2001-2005
Author(s):  
Irom Anil Singh ◽  
Aditi Jain ◽  
Pukhrambam Vedanti Devi ◽  
Tombing Niangneihching
Keyword(s):  
Chronic Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Kinase Inhibitors ◽  
Chronic Phase ◽  
Response To Treatment ◽  
P Value ◽  
Molecular Response ◽  
Optimal Response ◽  
Potent Drug

BACKGROUND Chronic myeloid leukemia (CML) accounts for 15 - 20 % of leukemia in adults worldwide. At present, the three tyrosine kinase inhibitors (TKI) imatinib, dasatinib, or nilotinib are accepted as the standard first-line treatment in chronic phase (CP). Nilotinib is a second generation TKI having faster and deeper response compared to imatinib. We wanted to see if the response achieved with nilotinib in the first three months could be translated into long term benefits when imatinib was given after 3 months. METHODS Newly diagnosed CML-CP patients were randomized into two arms. The patients on the first arm were given imatinib and in the second arm nilotinib was given for first 3 months. After three months nilotinib was switched over to imatinib. The molecular response was assessed in both arms at 3 months and 6 months. RESULTS Twenty-six patients in each arm were analysed. The optimal molecular response (QPCR <10 %) after 3 months was significantly higher in patients receiving nilotinib than imatinib (96.1 % vs 65.38 %; P < 0.0048). The optimal response after 6 months (QPCR < 1 %) was found to be more in the initial nilotinib arm than the initial imatinib arm (76.9 % vs 65.3 %; P - value = 0.35). CONCLUSIONS Patients on nilotinib arm did well even after switching to imatinib. It gives us an important platform for an economically backward country like India where the therapy with more potent drug like nilotinib is given in the initial three months or even six months. KEYWORDS Chronic Myeloid Leukaemia, Imatinib, Nilotinib, Optimal Response, Major Molecular Response

scholarly journals Molecular monitoring in CML: how deep? How often? How should it influence therapy?

Hematology ◽  
2018 ◽  
Vol 2018 (1) ◽  
pp. 168-176 ◽  
Author(s):  
Naranie Shanmuganathan ◽  
Timothy P. Hughes
Keyword(s):  
Messenger Rna ◽  
Kinase Inhibitors ◽  
Quantitative Polymerase Chain Reaction ◽  
Response Assessment ◽  
Molecular Response ◽  
Molecular Monitoring ◽  
Optimal Outcomes ◽  
Patients At Risk ◽  
Failure Depth ◽  
Treatment Free Remission

Abstract With the advent of tyrosine kinase inhibitors (TKIs), the goals of therapy in chronic myeloid leukemia (CML) are steadily shifting. Long-term disease control on TKI therapy has been the goal and expectation for most patients. More recently, treatment-free remission (TFR) has entered mainstream practice and is increasingly being adopted as the main goal of therapy. This therapeutic shift not only influences TKI selection but also, has necessitated the refinement and dissemination of highly sensitive and accurate molecular monitoring techniques. Measurement of BCR-ABL1 messenger RNA expression through reverse transcription quantitative polymerase chain reaction, reported according to the International Scale, has become the primary tool for response assessment in CML. Achieving specific time-dependent molecular milestones, as defined by global therapeutic guidelines, has been established as critical in maximizing optimal outcomes while identifying patients at risk of therapy failure. Depth and duration of a deep molecular response have become the new therapeutic targets in patients considered for TFR. Consequently, molecular monitoring in CML has become even more critical to ongoing response assessment, identifying patients with TKI resistance and poor drug adherence, and enabling TFR to be attempted safely and effectively.

scholarly journals Molecular Response of Chronic Myeloid Leukaemia Patients to Tyrosine Kinase Inhibitors

2020 ◽  
Vol 3 (02) ◽  
pp. 42-46
Author(s):  
Md. Imran Hossain ◽  
Md. Jaki Yamani ◽  
Md. Golzar Hossain ◽  
Mohammed Murad Hossain ◽  
Md. Rafiquzzaman Khan ◽  
...  
Keyword(s):  
Chronic Myeloid Leukaemia ◽  
Tyrosine Kinase ◽  
Myeloid Leukaemia ◽  
Tyrosine Kinase Inhibitors ◽  
Kinase Inhibitors ◽  
Occupational Status ◽  
Total Leukocyte Count ◽  
Molecular Diagnostic ◽  
Molecular Response ◽  
Female Ratio

Background: The goal of therapy in Chronic Myeloid Leukaemia (CML) with tyrosine kinase inhibitors (TKIs) has been to achieve the molecular responses, as measured by the reduction or elimination of BCR-ABLl transcript. Objective: The aim of the study is to observe the molecular response status of CML patients to TKIs. Methodology: This is an observational study and was conducted in the department of Haematology, Bangabandhu Sheikh Mujib Medical University (BSMMU) from February 2018 to January 2019. A total of 30 diagnosed CML patients on TKIs therapy was checked for quantitative BCR-ABL1 (by qRT-PCR) level. The laboratory monitoring of BCR-ABL1 RNA with RQ-PCR was likely become the model for successful molecular diagnostic monitoring of CML patients to both assess and prognosticate the efficacy of these targeted treatments. Result: Among 30 diagnosed CML patients, the mean age was found 36.1±11.7 years with range from 20 to 70 years. Males were predominant 19 (63.3%), male: female ratio was 1.7:1. Almost two third (63.3%) patients were found optimal (?10%) after 3-month BCR ABL. Age, sex, religion, marital status, occupational status, BMI, monthly income, number of family member, symptoms and sings were not statistically significant (p>0.05) when compared between BCR ABL optimal (?10%) and warning (>10%) group. Mean haemoglobin, total leukocyte count, platelet count, basophil, myelocyte and baseline BCR ABL1 were not statistically significant (p>0.05) however atypical cells was found statistically significant (p

scholarly journals Clonal evolution and clinical implications of genetic abnormalities in blastic transformation of chronic myeloid leukaemia

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Yotaro Ochi ◽  
Kenichi Yoshida ◽  
Ying-Jung Huang ◽  
Ming-Chung Kuo ◽  
Yasuhito Nannya ◽  
...  
Keyword(s):  
Chronic Myeloid Leukaemia ◽  
Myeloid Leukaemia ◽  
Kinase Inhibitors ◽  
Blast Crisis ◽  
Clonal Evolution ◽  
Chronic Phase ◽  
Genetic Alterations ◽  
Prognostic Impact ◽  
Genetic Abnormalities ◽  
Prior Use

AbstractBlast crisis (BC) predicts dismal outcomes in patients with chronic myeloid leukaemia (CML). Although additional genetic alterations play a central role in BC, the landscape and prognostic impact of these alterations remain elusive. Here, we comprehensively investigate genetic abnormalities in 136 BC and 148 chronic phase (CP) samples obtained from 216 CML patients using exome and targeted sequencing. One or more genetic abnormalities are found in 126 (92.6%) out of the 136 BC patients, including the RUNX1-ETS2 fusion and NBEAL2 mutations. The number of genetic alterations increase during the transition from CP to BC, which is markedly suppressed by tyrosine kinase inhibitors (TKIs). The lineage of the BC and prior use of TKIs correlate with distinct molecular profiles. Notably, genetic alterations, rather than clinical variables, contribute to a better prediction of BC prognosis. In conclusion, genetic abnormalities can help predict clinical outcomes and can guide clinical decisions in CML.

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