多重基因即時定量PCR法應用於接受Glivec治療的慢性骨髓性白血病患者療效與預後評估之前瞻性研究
Other Title
Multiplex quantitative PCR for the assessment of treatment
response and outcome prediction in chronic myelogenous leukemia
patients receiving Glivec treatment: a prospective study
response and outcome prediction in chronic myelogenous leukemia
patients receiving Glivec treatment: a prospective study
Date Issued
2004
Date
2004
Author(s)
唐季祿
DOI
922314B002177
Abstract
Chronic myelogenous leukemia (CML) is a clonal disorder of hematopoietic stem cell
characterized by t(9;22), which results in fusion of bcr and abl proto-oncogenes and expression
of BCR-ABL chimeric protein with abnormal tyrosine-kinase activity. Inhibition of this kinase
activity by STI571 (Glivec) resulted in 96% of hematological remission and 68% cytogenetic
remission in newly diagnosed CML. Molecular remission is possible in a few patients.
Quantification of t(9;22) carrying cells can be achieved by real-time quantitative RT-PCR
(RQ-PCR) assay that is highly sensitive for evaluating treatment response and minimal residual
disease (MRD). We had successfully developed one-step RQ-RT-PCR method that integrated
RT and PCR reaction in single tube. The assay can detect one abnormal CML cells among 10 5
normal cells (5-log sensitivity). Amplification of GAPDH control gene was done in separate
reaction. Since each PCR reaction has its own kinetics, separate PCR assay for target and
control genes can potentially result in inaccurate quantification. Multiplex PCR can further
minimize the laboratory procedure and increase the assay accuracy. Whether multiplex
RQ-RT-PCR can be applied in detecting MRD in CML has not yet been reported in the
literature.
Between June 2001 and October 2004, a total of 102 CML patients had been treated with
Imatinib in NTUH, including 49 patients in chronic phase (CP), 30 patients in accelerated phase
(AP), and 23 patients in blastic phase (BC). Fifty-two were male and 50 female and the median
age was 40 years old (ranges 3-80). The median interval from diagnosis to Imatinib treatment
was 16 months (0-120) and median Imatinib duration 19.4 months (1-45). Since July 2004, most
patients received Imatinib as first-line drug in early chronic phase.
Results: the complete hematological rate (CHR) was 98% in CP, 70% in AP and 35% in BC
patients (p < 0.001). Major cytogenetic remission (MCyR) was achieved in 70% of CP, 58% in
AP and 75% in BC patients (p=0.08). However, only a few patients could achieved molecular
remission as defined as MRD < -3.0log reduction: 19% in CP, 11% in AP and 17% in BC.
(p=0.341). The probability of leukemic progression-free survival (PFS) at 3 years was 93% for
CP, 51% for AP and 4% for BC (p<0.001). PFS was also correlated with cytogenetic response
and molecular response. For patients treated in CP or AP stagesand had achieved MRD <
-2.0log reduction, none had leukemic progression. Cytogenetic or molecular remission in BC
stage didn ’t prevent disease relapse and progression. Sequencing analysis showed that most of
them had acquired mutation at BCR-ABL kinase domain.
In conclusion, Imatinib was highly effective in CML-CP. For advanced stages, higher doses
of imatinib or combination with other drugs are needed to improve response and prevent
leukemic progression to improve survival. Sequential monitoring of cytogenetic and molecular
analysis is helpful in clinical decision-making. (The result was presented at the annual meeting
of the Taiwan Society of Hematology, March 26, 2005 and manuscript in preparation for
submission)
Subjects
acute myeloid leukemia
minimal residual leukemia
real-time RT-PCR
WT-1 gene
Publisher
臺北市：國立臺灣大學醫學院內科
Type
report
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