COMPARISON BETWEEN 8-COLOR MULTIPARAMETER FLOW CYTOMETRY AND NEXT-GENERATION SEQUENCING TO DETECT MINIMAL RESIDUAL DISEASE IN MULTIPLE MYELOMA PATIENTS WHO UNDERWENT AUTO-SCT
(Abstract release date: 05/19/16)
EHA Library. TAKAMATSU H. 06/09/16; 132842; E1293
Dr. HIROYUKI TAKAMATSU
Contributions
Contributions
Abstract
Abstract: E1293
Type: Eposter Presentation
Background
Autologous stem cell transplantation (ASCT) in conjunction with novel therapeutic drugs can dramatically improve response rates and the prognosis of patients with multiple myeloma (MM). However, most patients with MM are considered to be incurable, and relapse owing to minimal residual disease (MRD) is the main cause of death among these patients. Therefore, new technologies to assess deeper responses are required. Next-generation sequencing (NGS) and multiparameter flow cytometry (MFC) methods have been used to assess MRD. However, the lack of standardization of conventional MFC approaches has had a negative impact on its reproducibility. Recently, a next-generation MFC method (EuroFlow) has been developed by the EuroFlow Consortium and the International Myeloma Foundation (IMF) for a highly sensitive and standardized detection of MRD in MM.
Aims
To compare the prognostic value of MRD detection in autografts in MM between NGS and EuroFlow.
Methods
A total of 22 newly diagnosed MM patients who underwent ASCT were included in this study. Median age 60 (range 41-65); males 14, females 8; ISS 1 (n=4), 2 (n=12), 3 (n=6). 6 patients showed high-risk chromosomal abnormalities (t(4;14) (n=6), del17p (n=1)). The induction regimen was bortezomib-based chemotherapy. All patients received melphalan 200 mg/sqm conditioning regimen before ASCT. 18 of 22 (82%) patients received maintenance therapy using lenalidomide or thalidomide. The best response post-ASCT was as follows: 10sCR, 1CR, 9VGPR, 2PR. 22 autografts, one from each MM patient, were analyzed using EuroFlow and NGS methods. The EuroFlow method was based on a standardized lyse-wash-and-stain sample preparation protocol, the measurement of high numbers of cells (≥5x106 cells/tube) and an optimized 8-color, 2-tubes, antibody panel, for accurate identification of plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC): tube 1: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /CD117APC /CD81 APC-C750 ; and tube 2: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /cytoplasmic (Cy) Immunoglobulin (Ig) κAPC /CyIgλAPC-C750. NGS-based MRD assessment was performed using the immunosequencing platform (Adaptive Biotechnologies, South San Francisco, CA) (Martinez-Lopez et al Blood 2014).
Results
We identified abnormal plasma cells (aPC) in autografts based on multivariate analysis of individual cells from each patient (e.g. CD56+, CD19-, CyIgκ+, CD117+). For the MM MRD in autografts, the events from tube 1 and tube 2 were combined and a median of 5.6×106 (range: 1.3×106-11.7×106) events was acquired. 12 of 22 (55%) cases were MRD positive by 8-color MFC while 18 of 22 (82%) cases were MRD positive by NGS. The correlation of MRD level between 8-color MFC and NGS was relatively high (Fig. A). There was no significant difference in PFS between MRD negative cases by EuroFlow (MRDMFC (-)) and MRDMFC (+) cases (Fig. B) (P=0.508). However, MRD negative cases by NGS (MRDNGS (-)) tended to show better PFS than MRDNGS (+) (P=0.114) (Fig. C). There was no significant difference in overall survival between the MRD positive and negative groups.
Conclusion
In this comparison study of MRD assessment in autografts using Euroflow and NGS approaches, the NGS platform showed higher sensitivity and prognostic value than EuroFlow.
Session topic: E-poster
Keyword(s): Autologous hematopoietic stem cell transplantation, Minimal residual disease (MRD), Myeloma
Type: Eposter Presentation
Background
Autologous stem cell transplantation (ASCT) in conjunction with novel therapeutic drugs can dramatically improve response rates and the prognosis of patients with multiple myeloma (MM). However, most patients with MM are considered to be incurable, and relapse owing to minimal residual disease (MRD) is the main cause of death among these patients. Therefore, new technologies to assess deeper responses are required. Next-generation sequencing (NGS) and multiparameter flow cytometry (MFC) methods have been used to assess MRD. However, the lack of standardization of conventional MFC approaches has had a negative impact on its reproducibility. Recently, a next-generation MFC method (EuroFlow) has been developed by the EuroFlow Consortium and the International Myeloma Foundation (IMF) for a highly sensitive and standardized detection of MRD in MM.
Aims
To compare the prognostic value of MRD detection in autografts in MM between NGS and EuroFlow.
Methods
A total of 22 newly diagnosed MM patients who underwent ASCT were included in this study. Median age 60 (range 41-65); males 14, females 8; ISS 1 (n=4), 2 (n=12), 3 (n=6). 6 patients showed high-risk chromosomal abnormalities (t(4;14) (n=6), del17p (n=1)). The induction regimen was bortezomib-based chemotherapy. All patients received melphalan 200 mg/sqm conditioning regimen before ASCT. 18 of 22 (82%) patients received maintenance therapy using lenalidomide or thalidomide. The best response post-ASCT was as follows: 10sCR, 1CR, 9VGPR, 2PR. 22 autografts, one from each MM patient, were analyzed using EuroFlow and NGS methods. The EuroFlow method was based on a standardized lyse-wash-and-stain sample preparation protocol, the measurement of high numbers of cells (≥5x106 cells/tube) and an optimized 8-color, 2-tubes, antibody panel, for accurate identification of plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC): tube 1: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /CD117APC /CD81 APC-C750 ; and tube 2: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /cytoplasmic (Cy) Immunoglobulin (Ig) κAPC /CyIgλAPC-C750. NGS-based MRD assessment was performed using the immunosequencing platform (Adaptive Biotechnologies, South San Francisco, CA) (Martinez-Lopez et al Blood 2014).
Results
We identified abnormal plasma cells (aPC) in autografts based on multivariate analysis of individual cells from each patient (e.g. CD56+, CD19-, CyIgκ+, CD117+). For the MM MRD in autografts, the events from tube 1 and tube 2 were combined and a median of 5.6×106 (range: 1.3×106-11.7×106) events was acquired. 12 of 22 (55%) cases were MRD positive by 8-color MFC while 18 of 22 (82%) cases were MRD positive by NGS. The correlation of MRD level between 8-color MFC and NGS was relatively high (Fig. A). There was no significant difference in PFS between MRD negative cases by EuroFlow (MRDMFC (-)) and MRDMFC (+) cases (Fig. B) (P=0.508). However, MRD negative cases by NGS (MRDNGS (-)) tended to show better PFS than MRDNGS (+) (P=0.114) (Fig. C). There was no significant difference in overall survival between the MRD positive and negative groups.
Conclusion
In this comparison study of MRD assessment in autografts using Euroflow and NGS approaches, the NGS platform showed higher sensitivity and prognostic value than EuroFlow.
Session topic: E-poster
Keyword(s): Autologous hematopoietic stem cell transplantation, Minimal residual disease (MRD), Myeloma
Abstract: E1293
Type: Eposter Presentation
Background
Autologous stem cell transplantation (ASCT) in conjunction with novel therapeutic drugs can dramatically improve response rates and the prognosis of patients with multiple myeloma (MM). However, most patients with MM are considered to be incurable, and relapse owing to minimal residual disease (MRD) is the main cause of death among these patients. Therefore, new technologies to assess deeper responses are required. Next-generation sequencing (NGS) and multiparameter flow cytometry (MFC) methods have been used to assess MRD. However, the lack of standardization of conventional MFC approaches has had a negative impact on its reproducibility. Recently, a next-generation MFC method (EuroFlow) has been developed by the EuroFlow Consortium and the International Myeloma Foundation (IMF) for a highly sensitive and standardized detection of MRD in MM.
Aims
To compare the prognostic value of MRD detection in autografts in MM between NGS and EuroFlow.
Methods
A total of 22 newly diagnosed MM patients who underwent ASCT were included in this study. Median age 60 (range 41-65); males 14, females 8; ISS 1 (n=4), 2 (n=12), 3 (n=6). 6 patients showed high-risk chromosomal abnormalities (t(4;14) (n=6), del17p (n=1)). The induction regimen was bortezomib-based chemotherapy. All patients received melphalan 200 mg/sqm conditioning regimen before ASCT. 18 of 22 (82%) patients received maintenance therapy using lenalidomide or thalidomide. The best response post-ASCT was as follows: 10sCR, 1CR, 9VGPR, 2PR. 22 autografts, one from each MM patient, were analyzed using EuroFlow and NGS methods. The EuroFlow method was based on a standardized lyse-wash-and-stain sample preparation protocol, the measurement of high numbers of cells (≥5x106 cells/tube) and an optimized 8-color, 2-tubes, antibody panel, for accurate identification of plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC): tube 1: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /CD117APC /CD81 APC-C750 ; and tube 2: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /cytoplasmic (Cy) Immunoglobulin (Ig) κAPC /CyIgλAPC-C750. NGS-based MRD assessment was performed using the immunosequencing platform (Adaptive Biotechnologies, South San Francisco, CA) (Martinez-Lopez et al Blood 2014).
Results
We identified abnormal plasma cells (aPC) in autografts based on multivariate analysis of individual cells from each patient (e.g. CD56+, CD19-, CyIgκ+, CD117+). For the MM MRD in autografts, the events from tube 1 and tube 2 were combined and a median of 5.6×106 (range: 1.3×106-11.7×106) events was acquired. 12 of 22 (55%) cases were MRD positive by 8-color MFC while 18 of 22 (82%) cases were MRD positive by NGS. The correlation of MRD level between 8-color MFC and NGS was relatively high (Fig. A). There was no significant difference in PFS between MRD negative cases by EuroFlow (MRDMFC (-)) and MRDMFC (+) cases (Fig. B) (P=0.508). However, MRD negative cases by NGS (MRDNGS (-)) tended to show better PFS than MRDNGS (+) (P=0.114) (Fig. C). There was no significant difference in overall survival between the MRD positive and negative groups.
Conclusion
In this comparison study of MRD assessment in autografts using Euroflow and NGS approaches, the NGS platform showed higher sensitivity and prognostic value than EuroFlow.
Session topic: E-poster
Keyword(s): Autologous hematopoietic stem cell transplantation, Minimal residual disease (MRD), Myeloma
Type: Eposter Presentation
Background
Autologous stem cell transplantation (ASCT) in conjunction with novel therapeutic drugs can dramatically improve response rates and the prognosis of patients with multiple myeloma (MM). However, most patients with MM are considered to be incurable, and relapse owing to minimal residual disease (MRD) is the main cause of death among these patients. Therefore, new technologies to assess deeper responses are required. Next-generation sequencing (NGS) and multiparameter flow cytometry (MFC) methods have been used to assess MRD. However, the lack of standardization of conventional MFC approaches has had a negative impact on its reproducibility. Recently, a next-generation MFC method (EuroFlow) has been developed by the EuroFlow Consortium and the International Myeloma Foundation (IMF) for a highly sensitive and standardized detection of MRD in MM.
Aims
To compare the prognostic value of MRD detection in autografts in MM between NGS and EuroFlow.
Methods
A total of 22 newly diagnosed MM patients who underwent ASCT were included in this study. Median age 60 (range 41-65); males 14, females 8; ISS 1 (n=4), 2 (n=12), 3 (n=6). 6 patients showed high-risk chromosomal abnormalities (t(4;14) (n=6), del17p (n=1)). The induction regimen was bortezomib-based chemotherapy. All patients received melphalan 200 mg/sqm conditioning regimen before ASCT. 18 of 22 (82%) patients received maintenance therapy using lenalidomide or thalidomide. The best response post-ASCT was as follows: 10sCR, 1CR, 9VGPR, 2PR. 22 autografts, one from each MM patient, were analyzed using EuroFlow and NGS methods. The EuroFlow method was based on a standardized lyse-wash-and-stain sample preparation protocol, the measurement of high numbers of cells (≥5x106 cells/tube) and an optimized 8-color, 2-tubes, antibody panel, for accurate identification of plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC): tube 1: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /CD117APC /CD81 APC-C750 ; and tube 2: CD138BV421 /CD27BV510 / CD38FITC /CD56PE /CD45PerCP-Cy5.5 /CD19PE-Cy7 /cytoplasmic (Cy) Immunoglobulin (Ig) κAPC /CyIgλAPC-C750. NGS-based MRD assessment was performed using the immunosequencing platform (Adaptive Biotechnologies, South San Francisco, CA) (Martinez-Lopez et al Blood 2014).
Results
We identified abnormal plasma cells (aPC) in autografts based on multivariate analysis of individual cells from each patient (e.g. CD56+, CD19-, CyIgκ+, CD117+). For the MM MRD in autografts, the events from tube 1 and tube 2 were combined and a median of 5.6×106 (range: 1.3×106-11.7×106) events was acquired. 12 of 22 (55%) cases were MRD positive by 8-color MFC while 18 of 22 (82%) cases were MRD positive by NGS. The correlation of MRD level between 8-color MFC and NGS was relatively high (Fig. A). There was no significant difference in PFS between MRD negative cases by EuroFlow (MRDMFC (-)) and MRDMFC (+) cases (Fig. B) (P=0.508). However, MRD negative cases by NGS (MRDNGS (-)) tended to show better PFS than MRDNGS (+) (P=0.114) (Fig. C). There was no significant difference in overall survival between the MRD positive and negative groups.
Conclusion
In this comparison study of MRD assessment in autografts using Euroflow and NGS approaches, the NGS platform showed higher sensitivity and prognostic value than EuroFlow.
Session topic: E-poster
Keyword(s): Autologous hematopoietic stem cell transplantation, Minimal residual disease (MRD), Myeloma
{{ help_message }}
{{filter}}