PMID- 29133595
OWN - NLM
STAT- MEDLINE
DCOM- 20190111
LR  - 20190816
IS  - 1557-3125 (Electronic)
IS  - 1541-7786 (Linking)
VI  - 16
IP  - 2
DP  - 2018 Feb
TI  - Targeted Next-Generation Sequencing for Detecting MLL Gene Fusions in Leukemia.
PG  - 279-285
LID - 10.1158/1541-7786.MCR-17-0569 [doi]
AB  - Mixed lineage leukemia (MLL) gene rearrangements characterize approximately 70% 
      of infant and 10% of adult and therapy-related leukemia. Conventional clinical 
      diagnostics, including cytogenetics and fluorescence in situ hybridization (FISH) 
      fail to detect MLL translocation partner genes (TPG) in many patients. 
      Long-distance inverse (LDI)-PCR, the "gold standard" technique that is used to 
      characterize MLL breakpoints, is laborious and requires a large input of genomic 
      DNA (gDNA). To overcome the limitations of current techniques, a targeted 
      next-generation sequencing (NGS) approach that requires low RNA input was tested. 
      Anchored multiplex PCR-based enrichment (AMP-E) was used to rapidly identify a 
      broad range of MLL fusions in patient specimens. Libraries generated using Archer 
      FusionPlex Heme and Myeloid panels were sequenced using the Illumina platform. 
      Diagnostic specimens (n = 39) from pediatric leukemia patients were tested with 
      AMP-E and validated by LDI-PCR. In concordance with LDI-PCR, the AMP-E method 
      successfully identified TPGs without prior knowledge. AMP-E identified 10 
      different MLL fusions in the 39 samples. Only two specimens were discordant; 
      AMP-E successfully identified a MLL-MLLT1 fusion where LDI-PCR had failed to 
      determine the breakpoint, whereas a MLL-MLLT3 fusion was not detected by AMP-E 
      due to low expression of the fusion transcript. Sensitivity assays demonstrated 
      that AMP-E can detect MLL-AFF1 in MV4-11 cell dilutions of 10(-7) and transcripts 
      down to 0.005 copies/ng.Implications: This study demonstrates a NGS methodology 
      with improved sensitivity compared with current diagnostic methods for 
      MLL-rearranged leukemia. Furthermore, this assay rapidly and reliably identifies 
      MLL partner genes and patient-specific fusion sequences that could be used for 
      monitoring minimal residual disease. Mol Cancer Res; 16(2); 279-85. (c)2017 AACR.
CI  - (c)2017 American Association for Cancer Research.
FAU - Afrin, Sadia
AU  - Afrin S
AD  - The University of Queensland Diamantina Institute, Translational Research 
      Institute, Brisbane, Australia.
FAU - Zhang, Christine R C
AU  - Zhang CRC
AD  - The University of Queensland Diamantina Institute, Translational Research 
      Institute, Brisbane, Australia.
FAU - Meyer, Claus
AU  - Meyer C
AD  - Institute of Pharm. Biology/DCAL, Goethe-University, Frankfurt/Main, Germany.
FAU - Stinson, Caedyn L
AU  - Stinson CL
AD  - The University of Queensland Diamantina Institute, Translational Research 
      Institute, Brisbane, Australia.
FAU - Pham, Thy
AU  - Pham T
AD  - The University of Queensland Diamantina Institute, Translational Research 
      Institute, Brisbane, Australia.
FAU - Bruxner, Timothy J C
AU  - Bruxner TJC
AD  - Institute for Molecular Bioscience, The University of Queensland, Brisbane, 
      Australia.
FAU - Venn, Nicola C
AU  - Venn NC
AD  - Children's Cancer Institute, University of New South Wales, Sydney, Australia.
FAU - Trahair, Toby N
AU  - Trahair TN
AD  - Kids Cancer Centre, Sydney Children's Hospital, Sydney, Australia.
FAU - Sutton, Rosemary
AU  - Sutton R
AD  - Children's Cancer Institute, University of New South Wales, Sydney, Australia.
FAU - Marschalek, Rolf
AU  - Marschalek R
AD  - Institute of Pharm. Biology/DCAL, Goethe-University, Frankfurt/Main, Germany.
FAU - Fink, J Lynn
AU  - Fink JL
AD  - The University of Queensland Diamantina Institute, Translational Research 
      Institute, Brisbane, Australia.
FAU - Moore, Andrew S
AU  - Moore AS
AD  - The University of Queensland Diamantina Institute, Translational Research 
      Institute, Brisbane, Australia. andrew.moore@uq.edu.au.
AD  - Oncology Services Group, Children's Health Queensland Hospital and Health 
      Service, Brisbane, Australia.
AD  - UQ Child Health Research Centre, The University of Queensland, Brisbane, 
      Australia.
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20171113
PL  - United States
TA  - Mol Cancer Res
JT  - Molecular cancer research : MCR
JID - 101150042
RN  - 0 (KMT2A protein, human)
RN  - 0 (Reagent Kits, Diagnostic)
RN  - 149025-06-9 (Myeloid-Lymphoid Leukemia Protein)
RN  - EC 2.1.1.43 (Histone-Lysine N-Methyltransferase)
SB  - IM
MH  - Child
MH  - Child, Preschool
MH  - Cohort Studies
MH  - Female
MH  - *Gene Fusion
MH  - High-Throughput Nucleotide Sequencing/methods
MH  - Histone-Lysine N-Methyltransferase/*genetics
MH  - Humans
MH  - Infant
MH  - Infant, Newborn
MH  - Leukemia/diagnosis/*genetics
MH  - Male
MH  - Myeloid-Lymphoid Leukemia Protein/*genetics
MH  - Reagent Kits, Diagnostic
MH  - Sensitivity and Specificity
MH  - Sequence Analysis, DNA/*methods
EDAT- 2017/11/15 06:00
MHDA- 2019/01/12 06:00
CRDT- 2017/11/15 06:00
PHST- 2017/10/05 00:00 [received]
PHST- 2017/10/17 00:00 [revised]
PHST- 2017/10/30 00:00 [accepted]
PHST- 2017/11/15 06:00 [pubmed]
PHST- 2019/01/12 06:00 [medline]
PHST- 2017/11/15 06:00 [entrez]
AID - 1541-7786.MCR-17-0569 [pii]
AID - 10.1158/1541-7786.MCR-17-0569 [doi]
PST - ppublish
SO  - Mol Cancer Res. 2018 Feb;16(2):279-285. doi: 10.1158/1541-7786.MCR-17-0569. Epub 
      2017 Nov 13.