Rapid and Sensitive Detection of Calreticulin Type 1 and 2 Mutations by Real-Time Quantitative PCR
Tóm tắt
The majority of patients with JAK2 V617F-negative essential thrombocythemia or primary myelofibrosis harbor mutations involving the calreticulin (CALR) gene. These mutations are located in CALR exon 9 and lead to a frameshift with subsequent alteration of the CALR protein C-terminus. They have emerged as valuable molecular markers for the diagnosis of clonal myeloproliferative diseases. Although a variety of CALR mutations have been described, two mutations, denoted type 1 and type 2, account for around 85 % of cases. The type 1 mutation encompasses a 52 bp deletion and the type 2 mutation a 5 bp TTGTC insertion. This work describes the development and testing of quantitative real-time PCRs (qPCRs) for detecting these two mutations. The final type 1 CALR qPCR displayed a sensitivity of <0.1 % mutant alleles and the type 2 CALR qPCR had a sensitivity of <0.01 % mutant alleles. Additionally, two new CALR mutations are reported. These sensitive and specific qPCRs should be helpful in establishing the diagnosis and in monitoring minimal residual disease in patients during or after therapy.
Tài liệu tham khảo
de Keersmaecker K, Cools J. Chronic myeloproliferative disorders: a tyrosine kinase tale. Leukemia. 2006;20:200–5.
Vardiman J, Hyjek E. World health organization classification, evaluation, and genetics of the myeloproliferative neoplasm variants. Hematology Am Soc Hematol Educ Program. 2011;2011:250–6.
Savage N, George TI, Gotlib J. Myeloid neoplasms associated with eosinophilia and rearrangement of PDGFRA, PDGFRB, and FGFR1: a review. Int J Lab Hematol. 2013;35:491–500.
Jackson CC, Medeiros LJ, Miranda RN. 8p11 myeloproliferative syndrome: a review. Hum Pathol. 2010;41:461–76.
Klampfl T, Gisslinger H, Harutyunyan AS, Nivarthi H, Rumi E, Milosevic JD, et al. Somatic mutations of calreticulin in myeloproliferative neoplasms. N Engl J Med. 2013;369:2379–90.
Nangalia J, Massie CE, Baxter EJ, Nice FL, Gundem G, Wedge DC, et al. Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2. N Engl J Med. 2013;369:2391–405.
Tefferi A, Lasho TL, Finke C, Belachew AA, Wassie EA, Ketterling RP, et al. Type 1 vs type 2 calreticulin mutations in primary myelofibrosis: differences in phenotype and prognostic impact. Leukemia. 2014;28:1568–70.
Jovanovic JV, Ivey A, Vannucchi AM, Lippert E, Oppliger Leibundgut E, Cassinat B, et al. Establishing optimal quantitative-polymerase chain reaction assays for routine diagnosis and tracking of minimal residual disease in JAK2-V617F-associated myeloproliferative neoplasms: a joint European LeukemiaNet/MPN&MPNr-EuroNet (COST action BM0902) study. Leukemia. 2013;27:2032–9.
Chalandon Y, Passweg JR, Guglielmi C, Iacobelli S, Apperley J, Schaap NP, et al. Early administration of donor lymphocyte infusions upon molecular relapse after allogeneic hematopoietic stem cell transplantation for chronic myeloid leukemia: a study by the Chronic Malignancies Working Party of the EBMT. Haematologica. 2014;99:1492–8.
Burmeister T, Marschalek R, Schneider B, Meyer C, Gökbuget N, Schwartz S, et al. Monitoring minimal residual disease by quantification of genomic chromosomal breakpoint sequences in acute leukemias with MLL aberrations. Leukemia. 2006;20:451–7.
Little S. Amplification-refractory mutation system (ARMS) analysis of point mutations. Curr Protoc Hum Genet. 2001;Chapter 9:Unit 9.8:9.8.1–9.8.12.
Liu J, Huang S, Sun M, Liu S, Liu Y, Wang W, et al. An improved allele-specific PCR primer design method for SNP marker analysis and its application. Plant Methods. 2012;8:34.
van der Velden VH, Hochhaus A, Cazzaniga G, Szczepanski T, Gabert J, van Dongen JJ. Detection of minimal residual disease in hematologic malignancies by real-time quantitative PCR: principles, approaches, and laboratory aspects. Leukemia. 2003;17:1013–34.
Tefferi A, Thiele J, Vannucchi AM, Barbui T. An overview on CALR and CSF3R mutations and a proposal for revision of WHO diagnostic criteria for myeloproliferative neoplasms. Leukemia. 2014;28:1407–13.
Baccarani M, Deininger MW, Rosti G, Hochhaus A, Soverini S, Apperley JF, et al. European LeukemiaNet recommendations for the management of chronic myeloid leukemia: 2013. Blood. 2013;122:872–84.
Lange T, Edelmann A, Siebolts U, Krahl R, Nehring C, Jäkel N, et al. JAK2 p. V617F allele burden in myeloproliferative neoplasms one month after allogeneic stem cell transplantation significantly predicts outcome and risk of relapse. Haematologica. 2013;98:722–8.
Bar M, Radich J. Maintenance therapy with tyrosine kinase inhibitors after transplant in patients with chronic myeloid leukemia. J Natl Compr Canc Netw. 2013;11:308–15.
Zhang WW, Habeebu S, Sheehan AM, Naeem R, Hernandez VS, Dreyer ZE, et al. Molecular monitoring of 8p11 myeloproliferative syndrome in an infant. J Pediatr Hematol Oncol. 2009;31:879–83.
Barraco D, Carobolante F, Candoni A, Simeone E, Piccaluga P, Tabanelli V, et al. Complete and long-lasting cytologic and molecular remission of FIP1L1-PDGFRA-positive acute eosinophil myeloid leukaemia, treated with low-dose imatinib monotherapy. Eur J Haematol. 2014;92:541–5.
Cheah CY, Burbury K, Apperley JF, Huguet F, Pitini V, Gardembas M, et al. Patients with myeloid malignancies bearing PDGFRB fusion genes achieve durable long-term remissions with imatinib. Blood. 2014;123:3574–7.
Haslam K, Langabeer SE, Molloy K, McMullin MF, Conneally E. Assessment of CALR mutations in myelofibrosis patients, post-allogeneic stem cell transplantation.[letter]. Br J Haematol. 2014;166(5):800–2.
Lim KH, Lin HC, Chen CG, Wang WT, Chang YC, Chiang YH, et al. Rapid and sensitive detection of CALR exon 9 mutations using high-resolution melting analysis. Clin Chim Acta. 2015;440:133–9.
Chi J, Manoloukos M, Pierides C, Nicolaidou V, Nicolaou K, Kleopa M, et al. Calreticulin mutations in myeloproliferative neoplasms and new methodology for their detection and monitoring. Ann Hematol. 2015;94:399–408.
Yao QM, Zhou J, Gale RP, Li JL, Li LD, Li N, et al. A rapid, highly accurate method for quantifying CALR mutant allele burden in persons with myeloproliferative neoplasms. Hematology. 2015. doi:10.1179/1607845415y.0000000009
Jones AV, Ward D, Lyon M, Leung W, Callaway A, Chase A, et al. Evaluation of methods to detect CALR mutations in myeloproliferative neoplasms. Leuk Res. 2015;39:82–7.