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. 2020 Nov 25;9(12):3818.
doi: 10.3390/jcm9123818.

Assessment of Minimal Residual Disease by Next Generation Sequencing in Peripheral Blood as a Complementary Tool for Personalized Transplant Monitoring in Myeloid Neoplasms

Paula Aguirre-Ruiz  1 , Beñat Ariceta  1   2 , María Cruz Viguria  2   3 , María Teresa Zudaire  2   3 , Zuriñe Blasco-Iturri  1 , Patricia Arnedo  3 , Almudena Aguilera-Diaz  2   4 , Axier Jauregui  3 , Amagoia Mañú  1   2 , Felipe Prosper  2   4   5 , María Carmen Mateos  2   3 , Marta Fernández-Mercado  1   2   4 , María José Larráyoz  1   2 , Margarita Redondo  2   3 , María José Calasanz  1   2 , Iria Vázquez  1   2 , Eva Bandrés  2   3
Affiliations

Assessment of Minimal Residual Disease by Next Generation Sequencing in Peripheral Blood as a Complementary Tool for Personalized Transplant Monitoring in Myeloid Neoplasms

Paula Aguirre-Ruiz et al. J Clin Med. .

Abstract

Patients with myeloid neoplasms who relapsed after allogenic hematopoietic stem cell transplant (HSCT) have poor prognosis. Monitoring of chimerism and specific molecular markers as a surrogate measure of relapse is not always helpful; therefore, improved systems to detect early relapse are needed. We hypothesized that the use of next generation sequencing (NGS) could be a suitable approach for personalized follow-up post-HSCT. To validate our hypothesis, we analyzed by NGS, a retrospective set of peripheral blood (PB) DNA samples previously evaluated by high-sensitive quantitative PCR analysis using insertion/deletion polymorphisms (indel-qPCR) chimerism engraftment. Post-HCST allelic burdens assessed by NGS and chimerism status showed a similar time-course pattern. At time of clinical relapse in 8/12 patients, we detected positive NGS-based minimal residual disease (NGS-MRD). Importantly, in 6/8 patients, we were able to detect NGS-MRD at time points collected prior to clinical relapse. We also confirmed the disappearance of post-HCST allelic burden in non-relapsed patients, indicating true clinical specificity. This study highlights the clinical utility of NGS-based post-HCST monitoring in myeloid neoplasia as a complementary specific analysis to high-sensitive engraftment testing. Overall, NGS-MRD testing in PB is widely applicable for the evaluation of patients following HSCT and highly valuable to personalized early treatment intervention when mixed chimerism is detected.

Keywords: chimerism; hematopoietic stem cell transplant (HSCT); minimal residual disease (MRD); myeloid leukemia; next generation sequencing (NGS).

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Conflict of interest statement

The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Specificity of the NGS-MRD analysis in non-relapsed patients. Specific negative NGS-MRD confirms remission during MC decreased until CC is reached in both UPN7 (A) and UPN17 (B). Post-HSCT engraftment analysis by indel-qPCR results are plotted as percentage of receptor (Y-axis) over time shown as days post-HSCT (X-axis). Vertical dotted lines denote the NGS-analysis time points and the height bars represents VAF percentages; asterisk indicate NGS-MRD variants. (NGS = next generation sequencing; MRD = minimal residual disease; MC = mixed chimerism; CC = complete chimerism; HSCT = hematopoietic stem cell transplant; UPN = unique patient number; VAF = variant allele frequency).
Figure 2
Figure 2
NGS-MRD markers for relapse detection in patients that achieved complete chimerism. Relapsed patients showed a correlation of chimerism status and NGS-MRD during the monitoring of the disease course; MC increase and NGS-MRD variants were detected prior to clinical relapse. (A) In UPN1, negative NGS-MRD correlated with CC and two new variants were detected with the slight increase of MC even before positive NGS-MRD presence. (B) In UPN3, no complete clearance of all the variants was achieved even during CC, and NGS-MRD turned positive when a slight increase of MC was detected. Post-HSCT engraftment analysis by indel-qPCR results are plotted as percentage of receptor (Y-axis) over time shown as days post-HSCT (X-axis). Vertical dotted lines denote the NGS-analysis time points and the height bars represents VAF percentages; asterisk indicate NGS-MRD variants. (NGS = next generation sequencing; MRD = minimal residual disease; MC = mixed chimerism; CC = complete chimerism; HSCT = hematopoietic stem cell transplant; UPN = unique patient number; VAF = variant allele frequency).
Figure 3
Figure 3
NGS-MRD markers for relapse detection in relapsed patients with MC fluctuations. NGS-MRD during MC monitoring helps to anticipate clinical relapse. Detection of positive NGS-MRD anticipates relapse 220 days in UPN12 (A) and 40 days in UPN19 (B). Post-HSCT engraftment analysis by indel-qPCR results are plotted as percentage of receptor (Y-axis) over time shown as days post-HSCT (X-axis). Vertical dotted lines denote the NGS-analysis time points and the height bars represents VAF percentages; asterisk indicate NGS-MRD variants. (NGS = next generation sequencing; MRD = minimal residual disease; MC = mixed chimerism; CC = complete chimerism; HSCT = hematopoietic stem cell transplant; UPN = unique patient number; VAF = variant allele frequency).
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