Klin Padiatr 2018; 230(03): 175
DOI: 10.1055/s-0038-1645023
Top 7 Novel approaches and applications
Georg Thieme Verlag KG Stuttgart · New York

High-throughput sequencing of clonal TCR alpha genes rearrangements in pediatric T-lineage acute lymphoblastic leukemia

A Komkov
1   Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia
2   Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia
,
A Miroshnichenkova
1   Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia
,
G Nugmanov
2   Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia
,
Y Lebedev
2   Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia
,
Y Olshanskaya
1   Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia
,
M Maschan
1   Dmitry Rogachev National Medical Research Center of Pediatric Hematology, Oncology and Immunology, Moscow, Russia
,
I Mamedov
2   Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry RAS, Moscow, Russia
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Publikationsverlauf

Publikationsdatum:
08. Mai 2018 (online)

 
 

    Introduction:

    Relapses are still the major cause of poor outcome in therapy of T-lineage acute lymphoblastic leukemia (T-ALL) in children. Over 30% cases of relapse in T-ALL are accompanied by changing of the main clones. At the present time the most reliable markers for analysis of clonal structure of T-ALL are rearrangements of T-cell receptors (TCR) genes: TCR beta, gamma and delta. The purpose of this study is identification and characterization a new type of clonal markers based on previously unanalyzed in ALL rearrangements of TCR alpha locus.

    Material and Methods:

    Detection of TCR alpha genes rearrangements was based on high-throughput sequencing of amplicons obtained in series of multiplex PCR with genomic DNA from bone marrow and primers for all major V- and J-genes combination of TCR alpha locus.

    Results:

    We analyzed 75 samples of T-ALL in which we identified over 90 clonal rearrangements of TCR alpha locus. 15 samples contained rearrangements with clonal rate more than 40%, 37 samples contained 1 – 4 rearrangements with lower clonal rate (5 – 35%), 23 samples didn't contain any leukemia specific TCR alpha rearrangements.

    Conclusion:

    For the first time we evaluated frequency of clonal rearrangements of TCR alpha locus in pediatric T-ALL. This type of markers is presented in more than half of T-ALL cases what makes them fully applicable for analysis of clonal structure of T-ALL in both onset and relapse.

    Funding: Russian Science Foundation grant # 17 – 75 – 10113.


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