Clonality profile in relapsed precursor-B-ALL children by GeneScan and sequencing analyses. Consequences on minimal residual disease monitoring

Germanò, Giuseppe; Giudice, Laura del; Palatron, Silvia; Giarin, Emanuela; Cazzaniga, Giovanni; Biondi, Andrea; Basso, Giuseppe

doi:10.1038/sj.leu.2403008

Cited by 45 publications

(37 citation statements)

References 36 publications

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“…[37][38][39][40] However, in most patients at least one IG/TCR gene rearrangement is preserved throughout the disease course. [37][38][39][40][41] To reduce the risk of false-negative results, preferably two IG/TCR gene rearrangements should be used as MRD-PCR target. 8 This should be facilitated by supplementing the classical IGH, IGK-Kde, TCRD, and TCRG targets with 'new' MRD-PCR targets, such as Vd2-Ja and TCRB gene rearrangements.…”

Section: Tablementioning

confidence: 99%

Analysis of minimal residual disease in childhood acute lymphoblastic leukemia: comparison between RQ-PCR analysis of Ig/TcR gene rearrangements and multicolor flow cytometric immunophenotyping

et al. 2004

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Detection of minimal residual disease (MRD) in follow-up samples from patients with ALL is essential for evaluation of treatment response. We applied multicolor flow cytometry and real-time quantitative PCR (RQ-PCR) to compare MRD results in 71 follow-up samples from 22 children treated for ALL. When results obtained by flow cytometry and RQ-PCR were grouped into positive-negative categories, a significant level of agreement was found in 72% of samples (Po0.001). However, if a cutoff level of 0.01% was applied, the concordance was 89%. MRD could be quantified in 19 samples by both methods, showing a strong correlation (Po0.01). Nevertheless, MRD levels differed more than five-fold between both methods in 4/ 19 samples. In 20 (28%) samples, the two techniques showed discordant results. Most discordant results (17/20) were due to the limited sensitivity of flow cytometry analysis within the range 0.01-0.001%; remaining discordant results were due to the instable or subclonal IG/TCR gene rearrangements or a limited quantitative range of the applied RQ-PCR targets. Although concordant results could be obtained by flow cytometry and RQ-PCR analysis, MRD levels may differ. Therefore, MRD data obtained by these two techniques are not yet easily exchangeable.

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Section: Tablementioning

confidence: 99%

Analysis of minimal residual disease in childhood acute lymphoblastic leukemia: comparison between RQ-PCR analysis of Ig/TcR gene rearrangements and multicolor flow cytometric immunophenotyping

et al. 2004

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“…1,11 We assume an association with the advanced disease duration of most patients. This assumption is supported by our observation that those patients transplanted during CR1 had a median marker number of five, whereas those with advanced disease that were transplanted during CR2 or CR3 showed a median marker number of three (Mann-Whitney test, P ¼ 0.04).…”

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confidence: 99%

Immunoglobulin and T-cell receptor gene rearrangements as PCR-based targets are stable markers for monitoring minimal residual disease in acute lymphoblastic leukemia after stem cell transplantation

et al. 2009

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by defining robust cut-off values in a larger, well-defined (international) reference cohort. Such future studies may also allow determination of clinically relevant thresholds in heterogenous samples. Other advantages are the potential for automation and the relatively low costs. We conclude that this type of functional read-out for p53 could be a valuable contribution to diagnostic and therapeutic criteria, and should be further tested in the prospective clinical trials. Conflict of interestThe authors declare no conflict of interest. AcknowledgementsArnon P Kater is supported by a 'Veni' grant from ZonMw (The Netherlands Organization for Health Research and Development). We thank Ronald van Ree for critical reading of the paper and comments.

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“…Specifically, 83% of original clonal sequences identified at diagnosis were conserved at relapse, compared to 71% found in our previous study on precursor-B-ALL patients. 6 Furthermore, the proportion of patients with all clonal PCR targets conserved at relapse was significantly higher in T-ALL (63%) in comparison with precursor-B-ALL (43%). 6 Moreover, our data suggest that TCRB gene rearrangements are more prone to undergo clonal evolution phenomena as compared to TCRG, TCRD and incomplete DJH rearrangements.…”

Section: To the Editormentioning

confidence: 99%

“…NG001333) and used the same conditions as for the amplification of complete Vb-Jb rearrangements. 9 All clonal gene rearrangements identified by heteroduplex or GeneScan analyses 6,8 were sequenced by dye-terminator cycle sequencing kit on ABI Prism 310 (Applied Biosystems, Foster City, CA, USA). TCR and DJH gene segments were identified in comparison with published sequences using NCBI database nucleotide-nucleotide BLAST (http://www.ncbi.nlm.nih.gov/blast/) and IGMT/ junctionAnalysis database (http://imgt.cines.fr/textes/vquest/).…”

Section: To the Editormentioning

confidence: 99%

Comparative sequence analysis of incomplete DJH and TCR gene rearrangements in children with relapses of T-ALL

et al. 2005

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The detection of minimal residual disease (MRD) during the first\ud phase of treatment can predict outcome in childhood acute\ud lymphoblastic leukemia (ALL).1 Currently MRD detection in ALL\ud patients provides important information in order to assign a\ud tailored-treatment and the risk of an impending relapse. Nevertheless,\ud the major treatment failure in ALL occurs predominantly\ud in patients with T-cell ALL. This reflects especially a more\ud therapy-resistance and a slower clearance of blasts of T-ALL in\ud comparison with precursor-B-ALL.2 Therefore, the quantification\ud of early response to therapy and the monitoring of MRD\ud during and after treatment can greatly improve the outcome and\ud long-term quality of life of these patients. In childhood ALL,\ud detection of MRD with high sensitivity (ie 104, 105) can be\ud achieved by quantitative PCR methods (RQ-PCR) of rearranged\ud immunoglobulin (Ig) and T-cell receptor (TCR) genes.1,3 The\ud usefulness of these specific PCR targets should give attention to\ud the possible modification of Ig and TCR gene rearrangements that\ud could occur during the course of disease, due to continue activity\ud of the V(D)J recombinase enzymes in leukemic blast

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Clonality profile in relapsed precursor-B-ALL children by GeneScan and sequencing analyses. Consequences on minimal residual disease monitoring

Cited by 45 publications

References 36 publications

Analysis of minimal residual disease in childhood acute lymphoblastic leukemia: comparison between RQ-PCR analysis of Ig/TcR gene rearrangements and multicolor flow cytometric immunophenotyping

Analysis of minimal residual disease in childhood acute lymphoblastic leukemia: comparison between RQ-PCR analysis of Ig/TcR gene rearrangements and multicolor flow cytometric immunophenotyping

Immunoglobulin and T-cell receptor gene rearrangements as PCR-based targets are stable markers for monitoring minimal residual disease in acute lymphoblastic leukemia after stem cell transplantation

Comparative sequence analysis of incomplete DJH and TCR gene rearrangements in children with relapses of T-ALL

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