Minimal residual disease in multiple myeloma: defining the role of next generation sequencing and flow cytometry in routine diagnostic use

8601 Background: Although molecular CR in multiple myeloma (MM) can be assessed by allele-specific oligonucleotide (ASO)-PCR, this technique requires preparation of clonotype-specific primers for each individual which is laborious and time-consuming. The usage of the LymphoSIGHT method, a next-generation sequencing (NGS)-based platform, may overcome these challenges and increase sensitivity and specificity. We compared the LymphoSIGHT approach with ASO-qPCR for minimal residual disease (MRD) detection in autografts in the autologous peripheral blood stem cell transplantation (ASCT) setting. Methods: Seventeen Japanese patients with newly diagnosed MM who received various induction regimens prior to ASCT were retrospectively analyzed. All patients had achieved a PR or CR after ASCT. Bone marrow (BM) slides from 13 MM patients and fresh BM cells from 4 MM patients at diagnosis as well as autografts were obtained for DNA extraction. Using universal primer sets, we amplified IGH variable (V), diversity (D), and joining (J) gene segments, IGH-DJ, and IGK from genomic DNA. Amplified products were subjected to deep sequencing using NGS. Reads were analyzed using standardized algorithms for clonotype determination. Myeloma-specific clonotypes were identified for each patient based on their high frequency in BM samples. The presence of the myeloma clonotype was then assessed in follow-up samples. Results: MRD in autografts was detected in 6 of 17 (35%) by ASO-qPCR and 13 of 17 (76%) by NGS. When MRD was assessed by NGS, 6 MRD (+) cases received post-ASCT therapy while 4 MRD (-) cases and 7 MRD (+) cases were followed without post-ASCT therapy. The MRD (-) cases tended to show a better PFS than the MRD (+) cases with post-ASCT therapy (P = 0.26) and those without post-ASCT therapy (P = 0.09) although overall survival rates were comparable among the three groups. There was no difference in PFS between MRD (-) and MRD (+) cases when MRD was assessed by ASO-qPCR (P = 0.6). These studies will be extended in 30 additional MM patients, and results will be presented. Conclusions: MRD-negativity in autografts revealed by NGS may be more closely associated with durable remission of MM than that revealed by ASO-qPCR.

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Immunoglobulin Gene Clonality Analysis using Next-Generation Sequencing for Improved Minimal Residual Disease Detection with Significant Prognostic Value in Multiple Myeloma Patients

Journal of Molecular Diagnostics ◽

10.1016/j.jmoldx.2021.09.006 ◽

2021 ◽

Author(s):

Jihye Ha ◽

Hyeonah Lee ◽

Saeam Shin ◽

Hyunsoo Cho ◽

Haerim Chung ◽

...

Keyword(s):

Multiple Myeloma ◽

Next Generation Sequencing ◽

Minimal Residual Disease ◽

Prognostic Value ◽

Residual Disease ◽

Immunoglobulin Gene ◽

Next Generation ◽

Clonality Analysis ◽

Generation Sequencing ◽

Minimal Residual

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Comparison of next-generation sequencing (NGS) and next-generation flow (NGF) for minimal residual disease (MRD) assessment in multiple myeloma

Blood Cancer Journal ◽

10.1038/s41408-020-00377-0 ◽

2020 ◽

Vol 10 (10) ◽

Author(s):

Alejandro Medina ◽

Noemi Puig ◽

Juan Flores-Montero ◽

Cristina Jimenez ◽

M.-Eugenia Sarasquete ◽

...

Keyword(s):

Multiple Myeloma ◽

Next Generation Sequencing ◽

Minimal Residual Disease ◽

Cox Regression ◽

Residual Disease ◽

Next Generation ◽

Increased Risk ◽

Next Generation Sequencing Ngs ◽

Generation Sequencing ◽

Minimal Residual

Abstract Detecting persistent minimal residual disease (MRD) allows the identification of patients with an increased risk of relapse and death. In this study, we have evaluated MRD 3 months after transplantation in 106 myeloma patients using a commercial next-generation sequencing (NGS) strategy (LymphoTrack®), and compared the results with next-generation flow (NGF, EuroFlow). The use of different marrow pulls and the need of concentrating samples for NGS biased the applicability for MRD evaluation and favored NGF. Despite that, correlation between NGS and NGF was high (R2 = 0.905). The 3-year progression-free survival (PFS) rates by NGS and NGF were longer for undetectable vs. positive patients (NGS: 88.7% vs. 56.6%; NGF: 91.4% vs. 50%; p < 0.001 for both comparisons), which resulted in a 3-year overall survival (OS) advantage (NGS: 96.2% vs. 77.3%; NGF: 96.6% vs. 74.9%, p < 0.01 for both comparisons). In the Cox regression model, NGS and NGF negativity had similar results but favoring the latter in PFS (HR: 0.20, 95% CI: 0.09–0.45, p < 0.001) and OS (HR: 0.21, 95% CI: 0.06–0.75, p = 0.02). All these results reinforce the role of MRD detection by different strategies in patient prognosis and highlight the use of MRD as an endpoint for multiple myeloma treatment.

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Minimal residual disease undetectable by next-generation sequencing predicts improved outcome in CLL after chemoimmunotherapy

Blood ◽

10.1182/blood.2019001077 ◽

2019 ◽

Vol 134 (22) ◽

pp. 1951-1959 ◽

Cited By ~ 8

Author(s):

Philip A. Thompson ◽

Jaya Srivastava ◽

Christine Peterson ◽

Paolo Strati ◽

Jeffrey L. Jorgensen ◽

...

Keyword(s):

Flow Cytometry ◽

Next Generation Sequencing ◽

Minimal Residual Disease ◽

Residual Disease ◽

Progression Free Survival ◽

Next Generation ◽

Free Survival ◽

Generation Sequencing ◽

Minimal Residual ◽

Improved Outcome

Thompson and colleagues report that detection of minimal residual disease using next-generation sequencing, which is 2 orders of magnitude more sensitive than flow cytometry, is a much better predictor of progression-free survival.

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