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Clinical Trial
. 2021 Aug;39(3):293-303.
doi: 10.1002/hon.2864. Epub 2021 Mar 31.

Targeted locus amplification to detect molecular markers in mantle cell and follicular lymphoma

Elisa Genuardi  1 , Petra Klous  2 , Barbara Mantoan  1 , Daniela Drandi  1 , Martina Ferrante  1 , Federica Cavallo  1   3 , Beatrice Alessandria  1 , Irene Dogliotti  1   3 , Daniele Grimaldi  1   3 , Simone Ragaini  1   3 , Michele Clerico  1   3 , Mariella Lo Schirico  1 , Elona Saraci  4 , Mehmet Yilmaz  2 , Gian Maria Zaccaria  1 , Sergio Cortelazzo  5 , Umberto Vitolo  6 , Stefano Luminari  7   8 , Massimo Federico  8 , Mario Boccadoro  1   3 , Max van Min  2 , Erik Splinter  2 , Marco Ladetto  9 , Simone Ferrero  1   3
Affiliations
Clinical Trial

Targeted locus amplification to detect molecular markers in mantle cell and follicular lymphoma

Elisa Genuardi et al. Hematol Oncol. 2021 Aug.

Abstract

Minimal residual disease (MRD) monitoring by PCR methods is a strong and standardized predictor of clinical outcome in mantle cell lymphoma (MCL) and follicular lymphoma (FL). However, about 20% of MCL and 40% of FL patients lack a reliable molecular marker, being thus not eligible for MRD studies. Recently, targeted locus amplification (TLA), a next-generation sequencing (NGS) method based on the physical proximity of DNA sequences for target selection, identified novel gene rearrangements in leukemia. The aim of this study was to test TLA in MCL and FL diagnostic samples lacking a classical, PCR-detectable, t(11; 14) MTC (BCL1/IGH), or t(14; 18) major breakpoint region and minor cluster region (BCL2/IGH) rearrangements. Overall, TLA was performed on 20 MCL bone marrow (BM) or peripheral blood (PB) primary samples and on 20 FL BM, identifying a novel BCL1 or BCL2/IGH breakpoint in 16 MCL and 8 FL patients (80% and 40%, respectively). These new breakpoints (named BCL1-TLA and BCL2-TLA) were validated by ASO primers design and compared as MRD markers to classical IGH rearrangements in eight MCL: overall, MRD results by BCL1-TLA were superimposable (R Pearson = 0.76) to the standardized IGH-based approach. Moreover, MRD by BCL2-TLA reached good sensitivity levels also in FL and was predictive of a primary refractory case. In conclusion, this study offers the proof of principle that TLA is a promising and reliable NGS-based technology for the identification of novel molecular markers, suitable for further MRD analysis in previously not traceable MCL and FL patients.

Keywords: NGS; follicular lymphoma; mantle cell lymphoma; marker screening; minimal residual disease.

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Conflict of interest statement

Petra Klous, Mehmet Yilmaz, Max van Min, and Erik Splinter are Cergentis employees. The other authors declare no conflicts of interest.

Figures

FIGURE 1
FIGURE 1
Study workflow and results overview
FIGURE 2
FIGURE 2
Physical relationship among the translocated loci represented by Circos
FIGURE 3
FIGURE 3
Schematic representation of mated chromosomic regions compared to MTC, MBR, and MCR breakpoint genomic localization. MTC, major translocation cluster; CCND1, cyclin D1 gene; MBR, major breakpoint region; MCR, minor cluster region
FIGURE 4
FIGURE 4
Correlation between BCL1‐TLA and IGH quantification. Major discordances were described as positive or PNQ for BCL1‐TLA and negative for IGH (red dots), positive BCL1‐TLA versus negative IGH sample (green dots). Minor discordance resulted as positive for BCL1‐TLA and PNQ for IGH (orange dot)
FIGURE 5
FIGURE 5
Minimal residual disease (MRD) trends by BCL1‐TLA (red line) and IGH (black line) in “double markers” mantle cell lymphoma patients with available FU samples postdiagnosis (panels A–I); MRD trends by BCL2‐TLA (black line) in follicular lymphoma patients (panel L–M)
FIGURE 6
FIGURE 6
Targeted locust amplification 13 standard curve performances (panel A and B) and minimal residual disease trends in bone marrow (blue line) and peripheral blood (green line) follow ups targeting BCL1‐TLA (panel C)
See this image and copyright information in PMC

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