Molecular interrogation of genetic information has transformed our understanding of disease and is now routinely integrated into the workup and monitoring of hematological malignancies. In this article, a brief but comprehensive review is presented of state-of-the-art testing in hematological disease.
Methods
The primary medical literature and standard textbooks in the field were queried and reviewed to assess current practices and trends for molecular testing in hematopathology by disease.
Results
Pertinent materials were summarized under appropriate disease categories.
Conclusion
Molecular testing is well entrenched in the diagnostic and therapeutic pathways for hematological malignancies, with rapid growth and insights emerging following the integration of next-generation sequencing into the clinical workflow.
Gonon-DemoulianR., GoldmanJ.M., NicoliniF.E.. History of chronic myeloid leukemia: a paradigm in the treatment of cancer [in French].Bull Cancer.2014; 101(1): 56–67.
4.
GahrtonG.. Historical note on the discovery of the Philadelphia chromosome.Cancer Genet.2012; 205(6): 338–339.
5.
ZhenC., WangY.L.. Molecular monitoring of chronic myeloid leukemia: international standardization of BCR-ABL1 quantitation.J Mol Diagn.2013; 15(5): 556–564.
6.
LeonardD.G., ed. Molecular Pathology in Clinical Practice: Genetics.New York: Springer;2008.
7.
NashedA.L., RaoK.W., GulleyM.L.. Clinical applications of BCR-ABL molecular testing in acute leukemia.J Mol Diagn.2003; 5(2): 63–72.
8.
PaneF., FrigeriF., SindonaM.. Neutrophilic-chronic myeloid leukemia: a distinct disease with a specific molecular marker (BCR/ABL with C3/A2 junction).Blood.1996; 88(7): 2410–2414.
9.
O'BrienS.G., GuilhotF., LarsonR.A.; IRIS Investigators. Imatinib compared with interferon and low-dose cytarabine for newly diagnosed chronic-phase chronic myeloid leukemia.N Engl J Med.2003; 348(11): 994–1004.
10.
BranfordS., FletcherL., CrossN.C.. Desirable performance characteristics for BCR-ABL measurement on an international reporting scale to allow consistent interpretation of individual patient response and comparison of response rates between clinical trials.Blood.2008; 112(8): 3330–3338.
11.
PressR.D.. Major molecular response in CML patients treated with tyrosine kinase inhibitors: the paradigm for monitoring targeted cancer therapy.Oncologist.2010; 15(7): 744–749.
12.
FrankfurtO., LichtJ.D.. Ponatinib–a step forward in overcoming resistance in chronic myeloid leukemia.Clin Cancer Res.2013; 19(21): 5828–5834.
13.
GotlibJ., MaxsonJ.E., GeorgeT.I.. The new genetics of chronic neutrophilic leukemia and atypical CML: implications for diagnosis and treatment.Blood.2013; 122(10): 1707–1711.
14.
SteensmaD.P., DewaldG.W., LashoT.L.. The JAK2 V617F activating tyrosine kinase mutation is an infrequent event in both “atypical” myeloproliferative disorders and myelodysplastic syndromes.Blood.2005; 106(4): 1207–1209.
15.
ElliottM.A., HansonC.A., DewaldG.W.. WHO-defined chronic neutrophilic leukemia: a long-term analysis of 12 cases and a critical review of the literature.Leukemia.2005; 19(2): 313–317.
16.
MaxsonJ.E., GotlibJ., PollyeaD.A.. Oncogenic CSF3R mutations in chronic neutrophilic leukemia and atypical CML.N Engl J Med.2013; 368(19): 1781–1790.
17.
ElliottM.A., TefferiA.. Chronic neutrophilic leukemia 2014: update on diagnosis, molecular genetics, and management.Am J Hematol.2014; 89(6): 651–658.
18.
BaxterE.J., ScottL.M., CampbellP.J.; Cancer Genome Project. Acquired mutation of the tyrosine kinase JAK2 in human myeloproliferative disorders.Lancet.2005; 365(9464): 1054–1061.
19.
SzpurkaH., JankowskaA.M., MakishimaH.. Spectrum of mutations in RARS-T patients includes TET2 and ASXL1 mutations.Leuk Res.2010; 34(8): 969–973.
20.
SwerdlowE.A.. WHO Classification of Tumours of the Heamatopoietic and Lymphoid Tissues.4th ed.Lyon: IARC; 2008.
21.
PancrazziA., GuglielmelliP., PonzianiV.. A sensitive detection method for MPLW515L or MPLW515K mutation in chronic myeloproliferative disorders with locked nucleic acid-modified probes and real-time polymerase chain reaction.J Mol Diagn.2008; 10(5): 435–441.
22.
PikmanY., LeeB.H., MercherT.. MPLW515L is a novel somatic activating mutation in myelofibrosis with myeloid metaplasia.PLoS Med.2006; 3(7): e270.
23.
DingJ., KomatsuH., WakitaA.. Familial essential thrombocythemia associated with a dominant-positive activating mutation of the c-MPL gene, which encodes for the receptor for thrombopoietin.Blood.2004; 103(11): 4198–4200.
24.
CankovicM., WhiteleyL., HawleyR.C.. Clinical performance of JAK2 V617F mutation detection assays in a molecular diagnostics laboratory: evaluation of screening and quantitation methods.Am J Clin Pathol.2009; 132(5): 713–721.
25.
SteensmaD.P.. JAK2 V617F in myeloid disorders: molecular diagnostic techniques and their clinical utility: a paper from the 2005 William Beaumont Hospital Symposium on Molecular Pathology.J Mol Diagn.2006; 8(4): 397–411.
26.
CazzolaM., KralovicsR.. From Janus kinase 2 to calreticulin: the clinically relevant genomic landscape of myeloproliferative neoplasms.Blood.2014; 123(24): 3714–3719.
27.
KlampflT., GisslingerH., HarutyunyanA.S.. Somatic mutations of calreticulin in myeloproliferative neoplasms.N Engl J Med.2013; 369(25): 2379–2390.
28.
NangaliaJ., MassieC.E., BaxterE.J.. Somatic CALR mutations in myeloproliferative neoplasms with nonmutated JAK2.N Engl J Med.2013; 369(25): 2391–2405.
29.
GangatN., CaramazzaD., VaidyaR.. DIPSS plus: a refined Dynamic International Prognostic Scoring System for primary myelofibrosis that incorporates prognostic information from karyotype, platelet count, and transfusion status.J Clin Oncol.2011; 29(4): 392–397.
30.
KristensenT., VestergaardH., MøllerM.B.. Improved detection of the KIT D816V mutation in patients with systemic mastocytosis using a quantitative and highly sensitive real-time qPCR assay.J Mol Diagn.2011; 13(2): 180–188.
31.
TefferiA., VerstovsekS., PardananiA.. How we diagnose and treat WHO-defined systemic mastocytosis in adults.Haematologica.2008; 93(1): 6–9.
32.
LimK.H., TefferiA., LashoT.L.. Systemic mastocytosis in 342 consecutive adults: survival studies and prognostic factors.Blood.2009; 113(23): 5727–5736.
33.
PardananiA.. Systemic mastocytosis in adults: 2013 update on diagnosis, risk stratification, and management.Am J Hematol.2013; 88(7): 612–624.
34.
GotlibJ.. World Health Organization-defined eosinophilic disorders: 2014 update on diagnosis, risk stratification, and management.Am J Hematol.2014; 89(3): 325–337.
35.
CoolsJ., DeAngeloD.J., GotlibJ.. A tyrosine kinase created by fusion of the PDGFRA and FIP1L1 genes as a therapeutic target of imatinib in idiopathic hypereosinophilic syndrome.N Engl J Med.2003; 348(13): 1201–1214.
JacobyM.A., WalterM.J.. Detection of copy number alterations in acute myeloid leukemia and myelodysplastic syndromes.Expert Rev Mol Diagn.2012; 12(3): 253–264.
38.
LaiJ.L., PreudhommeC., ZandeckiM.. Myelodysplastic syndromes and acute myeloid leukemia with 17p deletion. An entity characterized by specific dysgranulopoïesis and a high incidence of P53 mutations.Leukemia.1995; 9(3): 370–381.
39.
GreenbergP., CoxC., LeBeauM.M.. International scoring system for evaluating prognosis in myelodysplastic syndromes.Blood.1997; 89(6): 2079–2088.
40.
SchanzJ., TüchlerH., SoléF.. New comprehensive cytogenetic scoring system for primary myelodysplastic syndromes (MDS) and oligoblastic acute myeloid leukemia after MDS derived from an international database merge.J Clin Oncol.2012; 30(8): 820–829.
41.
MalcovatiL., Hellström-LindbergE., BowenD.; European Leukemia Net. Diagnosis and treatment of primary myelodysplastic syndromes in adults: recommendations from the European LeukemiaNet.Blood.2013; 122(17): 2943–2964.
42.
BejarR.. Prognostic models in myelodysplastic syndromes.Hematology Am Soc Hematol Educ Program.2013; 2013: 504–510.
43.
PapaemmanuilE., GerstungM., MalcovatiL.; Chronic Myeloid Disorders Working Group of the International Cancer Genome Consortium. Clinical and biological implications of driver mutations in myelodysplastic syndromes.Blood.2013; 122(22): 3616–3627.
44.
BejarR., StevensonK., Abdel-WahabO.. Clinical effect of point mutations in myelodysplastic syndromes.N Engl J Med.2011; 364(26): 2496–2506.
45.
BravoG.M., LeeE., MerchanB.. Integrating genetics and epigenetics in myelodysplastic syndromes: advances in pathogenesis and disease evolution.Br J Haematol.2014; 166(5): 646–659.
46.
HaferlachT., NagataY., GrossmannV.. Landscape of genetic lesions in 944 patients with myelodysplastic syndromes.Leukemia.2014; 28(2): 241–247.
47.
FramptonG.M., FichtenholtzA., OttoG.A.. Development and validation of a clinical cancer genomic profiling test based on massively parallel DNA sequencing.Nat Biotechnol.2013; 31(11): 1023–1031.
Nestal de MoraesG., CastroC.P., SalustianoE.J.. The pterocarpanquinone LQB-118 induces apoptosis in acute myeloid leukemia cells of distinct molecular subtypes and targets FoxO3a and FoxM1 transcription factors.Int J Oncol.2014; 45(5): 1949–1958.
50.
GrimwadeD., WalkerH., OliverF.; The Medical Research Council Adult and Children's Leukaemia Working Parties. The importance of diagnostic cytogenetics on outcome in AML: analysis of 1,612 patients entered into the MRC AML 10 trial.Blood.1998; 92(7): 2322–2333.
51.
SlovakM.L., KopeckyK.J., CassilethP.A.. Karyotypic analysis predicts outcome of preremission and postremission therapy in adult acute myeloid leukemia: a Southwest Oncology Group/Eastern Cooperative Oncology Group study.Blood.2000; 96(13): 4075–4083.
52.
MrózekK., MarcucciG., NicoletD.. Prognostic significance of the European LeukemiaNet standardized system for reporting cytogenetic and molecular alterations in adults with acute myeloid leukemia.J Clin Oncol.2012; 30(36): 4515–4523.
53.
DöhnerH., EsteyE.H., AmadoriS.; European LeukemiaNet. Diagnosis and management of acute myeloid leukemia in adults: recommendations from an international expert panel, on behalf of the European LeukemiaNet.Blood.2010; 115(3): 453–474.
54.
DufourA., SchneiderF., MetzelerK.H.. Acute myeloid leukemia with biallelic CEBPA gene mutations and normal karyotype represents a distinct genetic entity associated with a favorable clinical outcome.J Clin Oncol.2010; 28(4): 570–577.
EsteyE.H.. Acute myeloid leukemia: 2013 update on risk-stratification and management.Am J Hematol.2013; 88(4): 318–327.
57.
BullingerL., DöhnerK., KranzR.. An FLT3 gene-expression signature predicts clinical outcome in normal karyotype AML.Blood.2008; 111(9): 4490–4495.
58.
StirewaltD.L., RadichJ.P.. The role of FLT3 in haematopoietic malignancies.Nat Rev Cancer.2003; 3(9): 650–665.
59.
TanA.Y., WestermanD.A., CarneyD.A.. Detection of NPM1 exon 12 mutations and FLT3-internal tandem duplications by high resolution melting analysis in normal karyotype acute myeloid leukemia.J Hematol Oncol.2008; 1: 10.
60.
ThiedeC., CreutzigE., ReinhardtD.. Different types of NPM1 mutations in children and adults: evidence for an effect of patient age on the prevalence of the TCTG-tandem duplication in NPM1-exon 12.Leukemia.2007; 21(2): 366–367.
61.
LinL.I., ChenC.Y., LinD.T.. Characterization of CEBPA mutations in acute myeloid leukemia: most patients with CEBPA mutations have biallelic mutations and show a distinct immunophenotype of the leukemic cells.Clin Cancer Res.2005; 11(4): 1372–1379.
62.
LeroyH., RoumierC., HuygheP.. CEBPA point mutations in hematological malignancies.Leukemia.2005; 19(3): 329–334.
63.
GreenC.L., KooK.K., HillsR.K.. Prognostic significance of CEBPA mutations in a large cohort of younger adult patients with acute myeloid leukemia: impact of double CEBPA mutations and the interaction with FLT3 and NPM1 mutations.J Clin Oncol.2010; 28(16): 2739–2747.
64.
NaoeT., KiyoiH.. Gene mutations of acute myeloid leukemia in the genome era.Int J Hematol.2013; 97(2): 165–174.
HaferlachC., BacherU., HaferlachT.. The inv(3)(q21q26)/t(3;3) (q21;q26) is frequently accompanied by alterations of the RUNX1, KRAS and NRAS and NF1 genes and mediates adverse prognosis both in MDS and in AML: a study in 39 cases of MDS or AML.Leukemia.2011; 25(5): 874–877.
67.
RogersH.J., VardimanJ.W., AnastasiJ.. Complex or monosomal karyotype and not blast percentage is associated with poor survival in acute myeloid leukemia and myelodysplastic syndrome patients with inv(3) (q21q26.2)/t(3;3)(q21;q26.2): a Bone Marrow Pathology Group study.Haematologica.2014; 99(5): 821–829.
68.
ReisP.P., WaldronL., GoswamiR.S.. mRNA transcript quantification in archival samples using multiplexed, color-coded probes.BMC Biotechnol.2011; 11: 46.
WangZ.Y., ChenZ.. Acute promyelocytic leukemia: from highly fatal to highly curable.Blood.2008; 111(5): 2505–2515.
71.
CairoliR., BeghiniA., GrilloG.. Prognostic impact of c-KIT mutations in core binding factor leukemias: an Italian retrospective study.Blood.2006; 107(9): 3463–3468.
72.
MoormanA.V., ChiltonL., WilkinsonJ.. A population-based cytogenetic study of adults with acute lymphoblastic leukemia [Erratum appears in Blood. 2010; 116(5): 1017].Blood.2010; 115(2): 206–214.
73.
HoelzerD., ThielE., LöfflerH.. Prognostic factors in a multicenter study for treatment of acute lymphoblastic leukemia in adults.Blood.1988; 71(1): 123–131.
74.
FieldingA.K.. How I treat Philadelphia chromosome-positive acute lymphoblastic leukemia.Blood.2010; 116(18): 3409–3417.
75.
HungerS.P., LuX., DevidasM.. Improved survival for children and adolescents with acute lymphoblastic leukemia between 1990 and 2005: a report from the Children's Oncology Group.J Clin Oncol.2012; 30(14): 1663–1669.
76.
HildenJ.M., DinndorfP.A., MeerbaumS.O.; Children's Oncology Group. Analysis of prognostic factors of acute lymphoblastic leukemia in infants: report on CCG 1953 from the Children's Oncology Group.Blood.2006; 108(2): 441–451.
77.
GrimaldiJ.C., MeekerT.C.. The t(5;14) chromosomal translocation in a case of acute lymphocytic leukemia joins the interleukin-3 gene to the immunoglobulin heavy chain gene.Blood.1989; 73(8): 2081–2085.
BorowitzM.J., HungerS.P., CarrollA.J.. Predictability of the t(1;19) (q23;p13) from surface antigen phenotype: implications for screening cases of childhood acute lymphoblastic leukemia for molecular analysis: a Pediatric Oncology Group study.Blood.1993; 82(4): 1086–1091.
80.
MullighanC.G.. Genome sequencing of lymphoid malignancies.Blood.2013; 122(24): 3899–3907.
81.
NachmanJ.B., HeeremaN.A., SatherH.. Outcome of treatment in children with hypodiploid acute lymphoblastic leukemia.Blood.2007; 110(4): 1112–1115.
82.
FerrandoA.A., NeubergD.S., DodgeR.K.. Prognostic importance of TLX1 (HOX11) oncogene expression in adults with T-cell acute lymphoblastic leukaemia.Lancet.2004; 363(9408): 535–536.
83.
TurkmenS., TimmermannB., BartelsG.. Involvement of the MLL gene in adult T-lymphoblastic leukemia.Genes Chromosomes Cancer.2012; 51(12): 1114–1124.
84.
BaggA.. Immunoglobulin and T-cell receptor gene rearrangements: minding your B's and T's in assessing lineage and clonality in neoplastic lymphoproliferative disorders.J Mol Diagn.2006; 8(4): 426–429.
85.
SandbergY., van Gastel-MolE.J., VerhaafB.. BIOMED-2 multiplex immunoglobulin/T-cell receptor polymerase chain reaction protocols can reliably replace Southern blot analysis in routine clonality diagnostics.J Mol Diagn.2005; 7(4): 495–503.
86.
Elenitoba-JohnsonK.S., BohlingS.D., MitchellR.S.. PCR analysis of the immunoglobulin heavy chain gene in polyclonal processes can yield pseudoclonal bands as an artifact of low B cell number.J Mol Diagn.2000; 2(2): 92–96.
87.
NihalM., MikkolaD., WoodG.S.. Detection of clonally restricted immunoglobulin heavy chain gene rearrangements in normal and lesional skin: analysis of the B cell component of the skin-associated lymphoid tissue and implications for the molecular diagnosis of cutaneous B cell lymphomas.J Mol Diagn.2000; 2(1): 5–10.
88.
GrodyW.W., NakamuraR.M., KiechleF.L.. eds. Molecular Diagnostics: Techniques and Applications for the Clinical Laboratory.San Diego, CA: Academic Press; 2010.
89.
WangS.A., WangL., HochbergE.P.. Low histologic grade follicular lymphoma with high proliferation index: morphologic and clinical features.Am J Surg Pathol.2005; 29(11): 1490–1496.
SwerdlowS.H.. Diagnosis of ‘double hit’ diffuse large B-cell lymphoma and B-cell lymphoma, unclassifiable, with features intermediate between DL-BCL and Burkitt lymphoma: when and how, FISH versus IHC.ASH Education Handbook.2014; (1): 90–99.
92.
ValeraA., BalagueO., ColomoL.. IG/MYC rearrangements are the main cytogenetic alteration in plasmablastic lymphomas.Am J Surg Pathol.2010; 34(11): 1686–1694.
HummelM., BentinkS., BergerH.; Molecular Mechanisms in Malignant Lymphomas Network Project of the Deutsche Krebshilfe. A biologic definition of Burkitt's lymphoma from transcriptional and genomic profiling.N Engl J Med.2006; 354(23): 2419–2430.
95.
HaralambievaE., BoermaE.J., van ImhoffG.W.. Clinical, immunophenotypic, and genetic analysis of adult lymphomas with morphologic features of Burkitt lymphoma.Am J Surg Pathol.2005; 29(8): 1086–1094.
96.
FerryJ.A.. Burkitt's lymphoma: clinicopathologic features and differential diagnosis.Oncologist.2006; 11(4): 375–383.
97.
CrisanD., ed. Hematopathology: Genomic Mechanisms of Neoplastic Diseases.New York: Humana Press;2010.
98.
SunT., NordbergM.L., CotelingamJ.D.. Fluorescence in situ hybridization: method of choice for a definitive diagnosis of mantle cell lymphoma.Am J Hematol.2003; 74(1): 78–84.
DierlammJ., BaensM., WlodarskaI.. The apoptosis inhibitor gene API2 and a novel 18q gene, MLT, are recurrently rearranged in the t(11;18) (q21;q21) associated with mucosa-associated lymphoid tissue lymphomas.Blood.1999; 93(11): 3601–3609.
101.
StreubelB., Simonitsch-KluppI., MüllauerL.. Variable frequencies of MALT lymphoma-associated genetic aberrations in MALT lymphomas of different sites.Leukemia.2004; 18(10): 1722–1726.
102.
HamoudiR.A., AppertA., YeH.. Differential expression of NF-kappaB target genes in MALT lymphoma with and without chromosome translocation: insights into molecular mechanism.Leukemia.2010; 24(8): 1487–1497.
103.
FabbriG., RasiS., RossiD.. Analysis of the chronic lymphocytic leukemia coding genome: role of NOTCH1 mutational activation.J Exp Med.2011; 208(7): 1389–1401.
104.
ArcainiL., ZibelliniS., BoveriE.. The BRAF V600E mutation in hairy cell leukemia and other mature B-cell neoplasms.Blood.2012; 119(1): 188–191.
TiacciE., SchiavoniG., ForconiF.. Simple genetic diagnosis of hairy cell leukemia by sensitive detection of the BRAF-V600E mutation.Blood.2012; 119(1): 192–195.
TreonS.P., HunterZ.R.. A new era for Waldenstrom macroglobulinemia: MYD88 L265P.Blood.2013; 121(22): 4434–4436.
109.
MurrayF., DarzentasN., HadzidimitriouA.. Stereotyped patterns of somatic hypermutation in subsets of patients with chronic lymphocytic leukemia: implications for the role of antigen selection in leukemogenesis.Blood.2008; 111(3): 1524–1533.
110.
RådestadE., WikellH., EngströmM.. Alpha/beta T-cell depleted grafts as an immunological booster to treat graft failure after hematopoietic stem cell transplantation with HLA-matched related and unrelated donors.J Immunol Res.2014; 2014: 578741.
LiangX., MeechS.J., OdomL.F.. Assessment of t(2;5)(p23;q35) translocation and variants in pediatric ALK+ anaplastic large cell lymphoma.Am J Clin Pathol.2004; 121(4): 496–506.
115.
RudolphC., BittnerC., FellerA.C.. Cytogenetic characteristics of a murine in vitro model for the human anaplastic large cell lymphoma (ALCL).Cytogenet Genome Res.2006; 114 (3-4): 292–295.
116.
DelgadoP., StarshakP., RaoN.. A comprehensive update on molecular and cytogenetic abnormalities in T-cell prolymphocytic leukemia (T-pll).J Assoc Genet Technol.2012; 38(4): 193–198.
117.
DeardenC.. How I treat prolymphocytic leukemia.Blood.2012; 120(3): 538–551.
118.
MandavaS., SonarR., AhmadF.. Cytogenetic and molecular characterization of a hepatosplenic T-cell lymphoma: report of a novel chromosomal aberration.Cancer Genet.2011; 204(2): 103–107.
119.
TsukasakiK., TsushimaH., YamamuraM.. Integration patterns of HTLV-I provirus in relation to the clinical course of ATL: frequent clonal change at crisis from indolent disease.Blood.1997; 89(3): 948–956.
