Detection and molecular monitoring of FIP1L1-PDGFRA-positive diseases by analysis of patient-specific genomic DNA fusion junctions
Detection and molecular monitoring of FIP1L1-PDGFRA-positive diseases by analysis of patient-specific genomic DNA fusion junctions
To evaluate current detection methods for FIP1L1-PDGFRA in hypereosinophilic syndrome (HES), we developed a means to rapidly amplify genomic break points. We screened 202 cases and detected genomic junctions in all samples previously identified as RT-PCR positive (n=43). Genomic fusions were amplified by single step PCR in all cases whereas only 22 (51%) were single step RT-PCR positive. Importantly, FIP1L1-PDGFRA was detected in two cases that initially tested negative by RTPCR or fluorescence in situ hybridization. Absolute quantitation of the fusion by real-time PCR from genomic DNA (gDNA) using patient-specific primer/probe combinations at presentation (n=13) revealed a 40-fold variation between patients (range, 0.027-1.1 FIP1L1-PDGFRA copies/haploid genome). In follow up samples, quantitative analysis of gDNA gave 1-2 log greater sensitivity than RQ-PCR of cDNA. Minimal residual disease assessment using gDNA showed that 11 of 13 patients achieved complete molecular response to imatinib within a median of 9 months (range, 3-17) of starting treatment, with a sensitivity of detection of up to 1 in 10 5. One case relapsed with an acquired D842V mutation. We conclude that detection of FIP1L1-PDGFRA from gDNA is a useful adjunct to standard diagnostic procedures and enables more sensitive follow up of positive cases after treatment. Leukemia (2009) 23, 332-339; doi:10.1038/leu.2008.309; published online 6 November 2008 Copyright (C) 2009 Nature Publishing Group
imatinib, FIP1L1-PDGFRA, HES, MRD
332-339
Score, J.
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Walz, C.
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Jovanovic, J.V.
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Jones, A.V.
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Waghorn, K.
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Hidalgo-Curtis, C.
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Lin, F.
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Grimwade, D.
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Grand, F.
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Reiter, A.
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Cross, N.C.P.
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February 2009
Score, J.
ea0db6ef-c17e-4915-b216-ac67c07b26b7
Walz, C.
03a7b61b-bebd-4510-8f08-ec6b65a36809
Jovanovic, J.V.
92ddc8f4-238a-4c32-8f06-f14fb1b5f294
Jones, A.V.
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Waghorn, K.
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Hidalgo-Curtis, C.
b492ffa5-2adb-4901-973f-72e2432b0ee0
Lin, F.
9067ee54-a57f-4432-b390-ec81fd8aa73c
Grimwade, D.
ffb05d2d-ba42-4841-a002-3c80c3b26961
Grand, F.
26daee41-f8f8-4bae-910f-89579532ea62
Reiter, A.
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Cross, N.C.P.
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Score, J., Walz, C., Jovanovic, J.V., Jones, A.V., Waghorn, K., Hidalgo-Curtis, C., Lin, F., Grimwade, D., Grand, F., Reiter, A. and Cross, N.C.P.
(2009)
Detection and molecular monitoring of FIP1L1-PDGFRA-positive diseases by analysis of patient-specific genomic DNA fusion junctions.
Leukemia, 23 (2), .
(doi:10.1038/leu.2008.309).
Abstract
To evaluate current detection methods for FIP1L1-PDGFRA in hypereosinophilic syndrome (HES), we developed a means to rapidly amplify genomic break points. We screened 202 cases and detected genomic junctions in all samples previously identified as RT-PCR positive (n=43). Genomic fusions were amplified by single step PCR in all cases whereas only 22 (51%) were single step RT-PCR positive. Importantly, FIP1L1-PDGFRA was detected in two cases that initially tested negative by RTPCR or fluorescence in situ hybridization. Absolute quantitation of the fusion by real-time PCR from genomic DNA (gDNA) using patient-specific primer/probe combinations at presentation (n=13) revealed a 40-fold variation between patients (range, 0.027-1.1 FIP1L1-PDGFRA copies/haploid genome). In follow up samples, quantitative analysis of gDNA gave 1-2 log greater sensitivity than RQ-PCR of cDNA. Minimal residual disease assessment using gDNA showed that 11 of 13 patients achieved complete molecular response to imatinib within a median of 9 months (range, 3-17) of starting treatment, with a sensitivity of detection of up to 1 in 10 5. One case relapsed with an acquired D842V mutation. We conclude that detection of FIP1L1-PDGFRA from gDNA is a useful adjunct to standard diagnostic procedures and enables more sensitive follow up of positive cases after treatment. Leukemia (2009) 23, 332-339; doi:10.1038/leu.2008.309; published online 6 November 2008 Copyright (C) 2009 Nature Publishing Group
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Published date: February 2009
Keywords:
imatinib, FIP1L1-PDGFRA, HES, MRD
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Local EPrints ID: 69894
URI: http://eprints.soton.ac.uk/id/eprint/69894
ISSN: 0887-6924
PURE UUID: 14848753-3dbd-4a6d-b45c-d682635b331c
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Date deposited: 09 Dec 2009
Last modified: 14 Mar 2024 02:46
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Author:
J. Score
Author:
C. Walz
Author:
J.V. Jovanovic
Author:
A.V. Jones
Author:
K. Waghorn
Author:
C. Hidalgo-Curtis
Author:
F. Lin
Author:
D. Grimwade
Author:
F. Grand
Author:
A. Reiter
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