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Prepublished online as a Blood First Edition Paper on December 5, 2002; DOI 10.1182/blood-2002-05-1357.
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Blood, 1 April 2003, Vol. 101, No. 7, pp. 2521-2528
CLINICAL OBSERVATIONS, INTERVENTIONS, AND THERAPEUTIC TRIALS
Quantitative real-time RT-PCR analysis of PML-RAR mRNA in
acute promyelocytic leukemia: assessment of prognostic significance in
adult patients from intergroup protocol 0129
Robert E. Gallagher,
Beow Y. Yeap,
Wanli Bi,
Kenneth J. Livak,
Nike Beaubier,
Sreenivas Rao,
Clara D. Bloomfield,
Frederick R. Appelbaum,
Martin S. Tallman,
James L. Slack, and
Cheryl L. Willman
From the Departments of Oncology and Medicine,
Montefiore Medical Center and Albert Einstein Cancer Center, Bronx, NY;
the Department of Medicine, Massachusetts General Hospital and Harvard
Medical School, Boston, MA; Applied Biosystems, Foster City, CA; The
Ohio State University Comprehensive Cancer Center, Columbus, OH; the
Fred Hutchinson Cancer Research Center, Seattle, WA; the Division of
Hematology/Oncology, Department of Medicine, Northwestern University
Medical School, Chicago, IL; the Department of Medicine, Roswell Park
Cancer Institute, Buffalo, NY; the Departments of Pathology and Cell
Biology and the University of New Mexico Cancer Center, University of
New Mexico, Albuquerque, NM.
The potential prognostic value of quantitative real-time reverse
transcription-polymerase chain reaction (RT-PCR [qrtPCR]) measurements of PML-RAR mRNA in acute promyelocytic leukemia was
retrospectively assessed before treatment and at 3 posttreatment intervals in 123 patients on intergroup protocol 0129. The primary measure was the PML-RARGAPDH normalized quotient (NQ),
that is, PML-RAR mRNA copies divided by
glyceraldehyde-3'-phosphate dehydrogenase (GAPDH) mRNA copies.
Only samples with more than 2.5 × 105 copies of the
housekeeping gene GAPDH mRNA (detection sensitivity exceeding
104) were considered NQ evaluable. With RNA from
low-density selected cells, paired peripheral blood (PB) and bone
marrow samples (n = 140) had comparable NQs
(P < .001). Before treatment, high NQ was
associated with short-form PML-RAR
(P < .001), but not with white blood cell
count or clinical outcome. Following treatment, NQ was lower
in all-trans retinoic acid-induced complete remission (CR)
than chemotherapy-induced CR (P = .018) and at
first test after consolidation chemotherapy
(P = .037). After consolidation chemotherapy, patients
with NQ exceeding 10 5 had 4.1-fold increased relapse risk
(P = .008); however, 73% of patients who experienced
relapse had NQ lower than 105. In the follow-up
period (FUP), any NQ exceeding 105 and
106 had 17.5-fold and 7.6-fold increased relapse risk,
respectively (P < .001), while no
gradation of relapse risk (approximately 18%) could be identified at
NQ lower than 106, including NQ. These
results indicate that qrtPCR monitoring of PML-RAR NQ can identify
patients at high risk of relapse and suggest that clinically practical
PB NQ monitoring at more frequent FUP intervals may improve predictive
accuracy for relapse or continuing CR in patients with persistent,
fluctuating minimal residual disease levels.
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