Reverse transcription polymerase chain reaction for the rearranged retinoic acid receptor α clarifies diagnosis and detects minimal residual disease in acute promyelocytic leukemia Journal Article

   

Authors:	Miller, W. H. Jr; Kakizuka, A.; Frankel, S. R.; Warrell, R. P. Jr; DeBlasio, A.; Levine, K.; Evans, R. M.; Dmitrovsky, E.
Article Title:	Reverse transcription polymerase chain reaction for the rearranged retinoic acid receptor α clarifies diagnosis and detects minimal residual disease in acute promyelocytic leukemia
Abstract:	The characteristic t(15;17) of acute promyelocytic leukemia (APL) fuses the retinoic acid receptor α (RAR-α) gene on chromosome 17 to a gene on chromosome 15 called PML, a putative transcription factor. This distinct translocation results in a fusion mRNA detected by Northern analysis. Two cDNAs have been isolated that differ in the extent of 3′ PML nucleic acid sequence contained. This study describes a reverse transcription polymerase chain reaction (RT-PCR) assay for the PML/RAR-α fusion transcript, which amplifies PML/RAR-α mRNA from APL cells with either reported breakpoint. DNA sequencing of the predominant RT-PCR products from 6 patients showed identical RAR-α exonic breakpoints and two PML breakpoints. This RT-PCR assay was positive in leukemic cells from 30/30 APL patients with the molecular rearrangement confirmed by cytogenetics or Northern analysis. In leukemic cells of patients with a morphologic diagnosis of APL lacking the t(15;17) by routine cytogenetics, a positive RT-PCR assay predicted clinical response to all-trans-retinoic acid (RA) therapy. Dilutional studies with leukemic cells that express (NB4) or do not express (HL-60) a PML/RAR-α fusion mRNA reveal that this RT-PCR assay detects the transcript from as little as 50 pg of total RNA. In APL cells from 5/6 patients treated with RA alone, a complete response by clinical and cytogenctic criteria accompanied a persistently positive RT-PCR assay. This preceded relapse by 1-6 months. RT-PCR for PML/RAR-α mRNA provides a more-sensitive test for the t(15;17) than routine cytogenetics or Northern analysis. This molecular rearrangement detected by RT-PCR best defines this RA-responsive malignancy. The RT-PCR assay for the PML/RAR-α transcript yields important diagnostic and prognostic information in the management of APL patients.
Keywords:	human cell; polymerase chain reaction; gene expression; transcription factor; cytogenetics; leukemia, promyelocytic, acute; gene rearrangement; molecular sequence data; diagnostic value; rna, messenger; leukemia cell; carrier proteins; acute myeloblastic leukemia; base sequence; dna sequence; chromosome aberrations; blotting, northern; retinoic acid; northern blotting; oligodeoxyribonucleotides; tretinoin; receptors, retinoic acid; chromosome translocation 17; chromosome disorders; rna directed dna polymerase; prognosis; human; priority journal; article; chromosome translocation 15; support, non-u.s. gov't; support, u.s. gov't, p.h.s.; rna-directed dna polymerase; translocation (genetics); retinoic acid binding protein
Journal Title:	Proceedings of the National Academy of Sciences of the United States of America
Volume:	89
Issue:	7
ISSN:	0027-8424
Publisher:	National Academy of Sciences
Date Published:	1992-04-01
Start Page:	2694
End Page:	2698
Language:	English
DOI:	10.1073/pnas.89.7.2694
PUBMED:	1372989
PROVIDER:	scopus
PMCID:	PMC48728
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-0026559655&doi=10.1073%2fpnas.89.7.2694&partnerID=40&md5=7e89a66f2b801273a0a132cbcb0e6d19
Notes:	Source: Scopus

https://synapse.mskcc.org/synapse/works/has_related_data_chk/150005