NEXT GENERATION FLOW CYTOMETRY IS A RELIABLE TECHNIQUE FOR DETECTING MINIMAL RESIDUAL DISEASE IN MULTIPLE MYELOMA PATIENTS IN COMPLETE REMISSION: A STUDY COMPARING SHORT VS LONG TERM REMISSION
(Abstract release date: 05/19/16)
EHA Library. Gozzetti A. 06/09/16; 132806; E1257
Dr. Alessandro Gozzetti
Contributions
Contributions
Abstract
Abstract: E1257
Type: Eposter Presentation
Background
4-6 colours flow cytometry has been validated in the past years as a powerful and feasible technique able to detect MRD in multiple myeloma patients thus furnishing a clinical and prognostic tool to the physician. Although its applicability and specificity can be high, it requires good quality bone marrow aspirate and expert and trained personnel for the analysis.However, recent results have confirmed that 4-6 colours flow-MRD has a lower sensitivity than molecular techniques such as allele-specific oligonucleotyde (ASO)-PCR and next generation sequencing (NGS). Recently, two 8 colours tubes panel developed by the EuroFlow Consortium showed to be able to detect MRD with an increased sensitivity and to be applicable as standardized method.
Aims
Complete remission (CR) is a prerequisite for long term responses, progression free survivals, and ultimately overall survivals and cure, but while many studies have looked at MRD status soon after autologous or allogeneic stem cell transplantation with flow or molecular techniques, little is known about long term remission patients (>5-10 years) and in particular if more sensitive techniques such as NGF or NGS can still detect minimal disease in those patients.
Methods
We tested the feasibility of NGF-MRD to CR MM patients with a lyse-wash-and-stain sample preparation protocol, staining an high numbers of bone marrow (BM) cells (≥2x106 cells/tube) with 2-tubes optimized 8-colours antibody panel, (OneFlow PCST e PCD BD Biosciences) for accurate identification of BM plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC). Patients with a CR/stringent CR were tested for MRD in BM and peripheral blood samples. Patients with 5-10 years of remission duration were analysed as well as patients with 2 to 4 years of remission. Clinical characteristics will be reported as well as treatment received.
Results
Applicability was 100% of patients studied. A preliminary result on the first 20 patients analysed showed an MRD + status in 12/20 patients predefined as CR. In particular, 2/20 patients did show residual disease also in the PB. Interestingly when studying patients with >5 years CR status, 1/5 did show an MRD+. A plan to analyse patients in CR is ongoing within the Tuscan Group of Multiple Myeloma (GTMM). The target is to test NGF-MRD in >100 CR patients comparing different treatments (auto vs chemo, vs new agents), age (<65 years vs >65 years), CR duration (<5 ys vs >5 ys).
Conclusion
Next generation flow is able to detect minimal residual disease in approximately 60% of predefined CR patients. The sensitivity can be quite comparable to the molecular techniques, but with an applicability largely superior. Patients with long term CR seem to have more chances to be MRD negative, although these findings nee to be demonstrated in a large cohort of patients. To confirm these data a study is currently ongoing of the GTMM and results will be presented.
Session topic: E-poster
Keyword(s): Flow cytometry, Minimal residual disease (MRD), Myeloma
Type: Eposter Presentation
Background
4-6 colours flow cytometry has been validated in the past years as a powerful and feasible technique able to detect MRD in multiple myeloma patients thus furnishing a clinical and prognostic tool to the physician. Although its applicability and specificity can be high, it requires good quality bone marrow aspirate and expert and trained personnel for the analysis.However, recent results have confirmed that 4-6 colours flow-MRD has a lower sensitivity than molecular techniques such as allele-specific oligonucleotyde (ASO)-PCR and next generation sequencing (NGS). Recently, two 8 colours tubes panel developed by the EuroFlow Consortium showed to be able to detect MRD with an increased sensitivity and to be applicable as standardized method.
Aims
Complete remission (CR) is a prerequisite for long term responses, progression free survivals, and ultimately overall survivals and cure, but while many studies have looked at MRD status soon after autologous or allogeneic stem cell transplantation with flow or molecular techniques, little is known about long term remission patients (>5-10 years) and in particular if more sensitive techniques such as NGF or NGS can still detect minimal disease in those patients.
Methods
We tested the feasibility of NGF-MRD to CR MM patients with a lyse-wash-and-stain sample preparation protocol, staining an high numbers of bone marrow (BM) cells (≥2x106 cells/tube) with 2-tubes optimized 8-colours antibody panel, (OneFlow PCST e PCD BD Biosciences) for accurate identification of BM plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC). Patients with a CR/stringent CR were tested for MRD in BM and peripheral blood samples. Patients with 5-10 years of remission duration were analysed as well as patients with 2 to 4 years of remission. Clinical characteristics will be reported as well as treatment received.
Results
Applicability was 100% of patients studied. A preliminary result on the first 20 patients analysed showed an MRD + status in 12/20 patients predefined as CR. In particular, 2/20 patients did show residual disease also in the PB. Interestingly when studying patients with >5 years CR status, 1/5 did show an MRD+. A plan to analyse patients in CR is ongoing within the Tuscan Group of Multiple Myeloma (GTMM). The target is to test NGF-MRD in >100 CR patients comparing different treatments (auto vs chemo, vs new agents), age (<65 years vs >65 years), CR duration (<5 ys vs >5 ys).
Conclusion
Next generation flow is able to detect minimal residual disease in approximately 60% of predefined CR patients. The sensitivity can be quite comparable to the molecular techniques, but with an applicability largely superior. Patients with long term CR seem to have more chances to be MRD negative, although these findings nee to be demonstrated in a large cohort of patients. To confirm these data a study is currently ongoing of the GTMM and results will be presented.
Session topic: E-poster
Keyword(s): Flow cytometry, Minimal residual disease (MRD), Myeloma
Abstract: E1257
Type: Eposter Presentation
Background
4-6 colours flow cytometry has been validated in the past years as a powerful and feasible technique able to detect MRD in multiple myeloma patients thus furnishing a clinical and prognostic tool to the physician. Although its applicability and specificity can be high, it requires good quality bone marrow aspirate and expert and trained personnel for the analysis.However, recent results have confirmed that 4-6 colours flow-MRD has a lower sensitivity than molecular techniques such as allele-specific oligonucleotyde (ASO)-PCR and next generation sequencing (NGS). Recently, two 8 colours tubes panel developed by the EuroFlow Consortium showed to be able to detect MRD with an increased sensitivity and to be applicable as standardized method.
Aims
Complete remission (CR) is a prerequisite for long term responses, progression free survivals, and ultimately overall survivals and cure, but while many studies have looked at MRD status soon after autologous or allogeneic stem cell transplantation with flow or molecular techniques, little is known about long term remission patients (>5-10 years) and in particular if more sensitive techniques such as NGF or NGS can still detect minimal disease in those patients.
Methods
We tested the feasibility of NGF-MRD to CR MM patients with a lyse-wash-and-stain sample preparation protocol, staining an high numbers of bone marrow (BM) cells (≥2x106 cells/tube) with 2-tubes optimized 8-colours antibody panel, (OneFlow PCST e PCD BD Biosciences) for accurate identification of BM plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC). Patients with a CR/stringent CR were tested for MRD in BM and peripheral blood samples. Patients with 5-10 years of remission duration were analysed as well as patients with 2 to 4 years of remission. Clinical characteristics will be reported as well as treatment received.
Results
Applicability was 100% of patients studied. A preliminary result on the first 20 patients analysed showed an MRD + status in 12/20 patients predefined as CR. In particular, 2/20 patients did show residual disease also in the PB. Interestingly when studying patients with >5 years CR status, 1/5 did show an MRD+. A plan to analyse patients in CR is ongoing within the Tuscan Group of Multiple Myeloma (GTMM). The target is to test NGF-MRD in >100 CR patients comparing different treatments (auto vs chemo, vs new agents), age (<65 years vs >65 years), CR duration (<5 ys vs >5 ys).
Conclusion
Next generation flow is able to detect minimal residual disease in approximately 60% of predefined CR patients. The sensitivity can be quite comparable to the molecular techniques, but with an applicability largely superior. Patients with long term CR seem to have more chances to be MRD negative, although these findings nee to be demonstrated in a large cohort of patients. To confirm these data a study is currently ongoing of the GTMM and results will be presented.
Session topic: E-poster
Keyword(s): Flow cytometry, Minimal residual disease (MRD), Myeloma
Type: Eposter Presentation
Background
4-6 colours flow cytometry has been validated in the past years as a powerful and feasible technique able to detect MRD in multiple myeloma patients thus furnishing a clinical and prognostic tool to the physician. Although its applicability and specificity can be high, it requires good quality bone marrow aspirate and expert and trained personnel for the analysis.However, recent results have confirmed that 4-6 colours flow-MRD has a lower sensitivity than molecular techniques such as allele-specific oligonucleotyde (ASO)-PCR and next generation sequencing (NGS). Recently, two 8 colours tubes panel developed by the EuroFlow Consortium showed to be able to detect MRD with an increased sensitivity and to be applicable as standardized method.
Aims
Complete remission (CR) is a prerequisite for long term responses, progression free survivals, and ultimately overall survivals and cure, but while many studies have looked at MRD status soon after autologous or allogeneic stem cell transplantation with flow or molecular techniques, little is known about long term remission patients (>5-10 years) and in particular if more sensitive techniques such as NGF or NGS can still detect minimal disease in those patients.
Methods
We tested the feasibility of NGF-MRD to CR MM patients with a lyse-wash-and-stain sample preparation protocol, staining an high numbers of bone marrow (BM) cells (≥2x106 cells/tube) with 2-tubes optimized 8-colours antibody panel, (OneFlow PCST e PCD BD Biosciences) for accurate identification of BM plasma cells (PCs) and discrimination between phenotypically aberrant (aPC) and normal PC (nPC). Patients with a CR/stringent CR were tested for MRD in BM and peripheral blood samples. Patients with 5-10 years of remission duration were analysed as well as patients with 2 to 4 years of remission. Clinical characteristics will be reported as well as treatment received.
Results
Applicability was 100% of patients studied. A preliminary result on the first 20 patients analysed showed an MRD + status in 12/20 patients predefined as CR. In particular, 2/20 patients did show residual disease also in the PB. Interestingly when studying patients with >5 years CR status, 1/5 did show an MRD+. A plan to analyse patients in CR is ongoing within the Tuscan Group of Multiple Myeloma (GTMM). The target is to test NGF-MRD in >100 CR patients comparing different treatments (auto vs chemo, vs new agents), age (<65 years vs >65 years), CR duration (<5 ys vs >5 ys).
Conclusion
Next generation flow is able to detect minimal residual disease in approximately 60% of predefined CR patients. The sensitivity can be quite comparable to the molecular techniques, but with an applicability largely superior. Patients with long term CR seem to have more chances to be MRD negative, although these findings nee to be demonstrated in a large cohort of patients. To confirm these data a study is currently ongoing of the GTMM and results will be presented.
Session topic: E-poster
Keyword(s): Flow cytometry, Minimal residual disease (MRD), Myeloma
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