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Published on: March 5, 2018
Quantification of minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL) using amplicon-fusion-site
Axel Weber1, Sylvia Taube, Udo Zur Stadt
1Department of Pediatric Oncology, Hematology and Hemostaseology, Children's Hospital, University of Leipzig, Leipzig, Germany. axel.weber@humangenetik.med.uni-giessen.de.
Abstract:
The amplification of putative oncogenes is a common finding within the genome of various cancer types. Identification and further targeting of specific junction sites within the sequence of genomic amplicons (amplicon fusion sites, AFS) by PCR (AFS-PCR) is suitable for quantification of minimal residual disease (MRD). This approach has recently been developed and described for MYCN amplified neuroblastomas. To compare AFS-PCR directly to routinely used MRD diagnostic strategies, we mapped the amplified genomic regions (ampGR) of an iAMP21-amplicon in high resolution of a patient with acute lymphoblastic leukemia (ALL). Successfully, we established AFS-PCR covering junction sites between ampGR within the iAMP21-amplicon. Quantification of MRD by AFS-PCR was directly comparable to IgH/TCR based real time quantitative PCR and fluorescence activated cell sorting (FACS) analysis in consecutive bone marrow (BM) specimens. Our data give an additional proof of concept of AFS-PCR for quantification of MRD. The method could be taken into account for ALL patients with genomic amplifications as alternative MRD diagnostic, if no or qualitatively poor Ig/TCR-PCRs are available.
Insights
Amplicon fusion site PCR (AFS-PCR) effectively quantifies minimal residual disease (MRD) in acute lymphoblastic leukemia (ALL). This method offers a comparable alternative to existing strategies, especially when other tests are unavailable.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Gene amplification is common in cancers, including oncogenes.
- Minimal residual disease (MRD) monitoring is crucial for cancer treatment efficacy.
- Amplicon fusion site PCR (AFS-PCR) is a novel method for MRD quantification.
Purpose of the Study:
- To establish and validate AFS-PCR for MRD quantification in acute lymphoblastic leukemia (ALL).
- To compare the efficacy of AFS-PCR against established MRD diagnostic methods.
Main Methods:
- High-resolution mapping of amplified genomic regions (ampGR) in an iAMP21-amplicon.
- Development of AFS-PCR targeting junction sites within the iAMP21-amplicon.
- Comparative analysis of AFS-PCR with IgH/TCR real-time quantitative PCR and FACS in bone marrow specimens.
Main Results:
- Successfully established AFS-PCR for the iAMP21-amplicon.
- AFS-PCR demonstrated comparable MRD quantification to IgH/TCR qPCR and FACS.
- Validated AFS-PCR as a viable method for MRD assessment.
Conclusions:
- AFS-PCR is a reliable method for quantifying MRD in ALL patients with genomic amplifications.
- AFS-PCR serves as a valuable alternative diagnostic tool when Ig/TCR-PCRs are unavailable or of poor quality.
