Cell sorting enables interphase fluorescencein situhybridization detection of lowBCR-ABL1producing stem cells in chronic myeloid leukaemia patients beyond deep molecular remission
| Autor: | Peter Hokland, Charlotte Christie Petersen, Hans Beier Ommen, Line Nederby, Charlotte Guldborg Nyvold, Anne Stidsholt Roug, Peter Buur van Kooten Niekerk, Eigil Kjeldsen |
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| Rok vydání: | 2013 |
| Předmět: |
Adult
Male Neoplasm Residual Fusion Proteins bcr-abl CD34 Biology Real-Time Polymerase Chain Reaction Adult Aged Aged 80 and over Female Flow Cytometry Fusion Proteins bcr-abl Humans In Situ Hybridization Fluorescence Interphase Leukemia Myelogenous Chronic BCR-ABL Positive Male Middle Aged Neoplasm Residual Real-Time Polymerase Chain Reaction Cytogenetics Quantitative polymerase chain reaction Leukemia Myelogenous Chronic BCR-ABL Positive hemic and lymphatic diseases medicine Humans Progenitor cell Interphase In Situ Hybridization Fluorescence Aged Aged 80 and over medicine.diagnostic_test Minimal residual disease Hematology Middle Aged Cell sorting Flow Cytometry Molecular biology Reverse transcription polymerase chain reaction Real-time polymerase chain reaction Fluorescence-activated cell sorting Female Stem cell Chronic myeloid leukaemia stem cells Fluorescence in situ hybridization |
| Zdroj: | van Kooten Niekerk, P B, Petersen, C C, Nyvold, C G, Ommen, H B, Roug, A S, Nederby, L, Hokland, P & Kjeldsen, E 2014, ' Cell sorting enables interphase fluorescence in situ hybridization detection of low BCR-ABL1 producing stem cells in chronic myeloid leukaemia patients beyond deep molecular remission ', British Journal of Haematology, vol. 164, no. 1, pp. 53-60 . https://doi.org/10.1111/bjh.12589 |
| ISSN: | 0007-1048 |
| DOI: | 10.1111/bjh.12589 |
| Popis: | Summary: The exact disease state of chronic myeloid leukaemia (CML) patients in deep molecular remission is unknown, because even the most sensitive quantitative reverse transcription polymerase chain reaction (qPCR) methods cannot identify patients prone to relapse after treatment withdrawal. To elucidate this, CD34 + stem cell and progenitor cell subpopulations were isolated by fluorescence-activated cell sorting (FACS), and their content of residual Philadelphia positive (Ph +) cells was evaluated in 17 CML patients (major molecular response, n = 6; 4-log reduction in BCR-ABL1 expression (MR 4), n = 11) using both sensitive qPCR and interphase fluorescence in situ hybridization (iFISH). Despite evaluating fewer cells, iFISH proved superior to mRNA-based qPCR in detecting residual Ph + stem cells (P = 0·005), and detected Ph + stem- and progenitor cells in 9/10 patients at frequencies of 2-14%. Moreover, while all qPCR + samples also were iFISH +, 9/33 samples were qPCR-/iFISH +, including all positive samples from MR 4 patients. Our findings show that residual Ph + cells are low BCR-ABL1 producers, and that DNA-based methods are required to assess the content of persisting Ph + stem cells in these patients. This approach demonstrates a clinically applicable manner of assessing residual disease at the stem cell level in CML patients in MR 4, and may enable early and safe identification of candidates for tyrosine kinase inhibitor withdrawal. |
| Databáze: | OpenAIRE |
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