Characterization of Immunophenotypic Aberrancies with Respect to Common Fusion Transcripts in B-Cell Precursor Acute Lymphoblastic Leukemia: A Report of 986 Indian Patients

Dikshat Gopal Gupta, Neelam Varma, Shano Naseem, Man Updesh Singh Sachdeva, Parveen Bose, Jogeshwar Binota, Ashish Kumar, Minakshi Gupta, Palak Rana, Preeti Sonam, Pankaj Malhotra, Amita Trehan, Alka Khadwal, Subhash Varma, Dikshat Gopal Gupta, Neelam Varma, Shano Naseem, Man Updesh Singh Sachdeva, Parveen Bose, Jogeshwar Binota, Ashish Kumar, Minakshi Gupta, Palak Rana, Preeti Sonam, Pankaj Malhotra, Amita Trehan, Alka Khadwal, Subhash Varma

Abstract

Objective: Based on the immunophenotype, acute lymphoblastic leukemia (ALL) can be categorized into B-cell or T-cell lineages. B-cell precursor ALL (BCP-ALL) cases show various genetic/molecular abnormalities, and varying frequencies of chimeric fusion transcripts in BCP-ALL cases are reported from different parts of the world. We studied the immunophenotypic aberrancy profiles of a large number of BCP-ALL cases with respect to various common chimeric fusion transcripts.

Materials and methods: Flow cytometric immunophenotyping and multiplex reverse-transcription polymerase chain reaction assays were performed for 986 BCP-ALL cases.

Results: Among 986 BCP-ALL cases, the incidence of various fusion transcripts was 38.36% in adult cases and 20.68% in pediatric cases. Adult BCP-ALL patients with t(9;22)(BCR-ABL1) fusion transcripts and expression of aberrant myeloid markers were significantly older at presentation (p=0.0218) with male preponderance (p=0.0246) compared to those without aberrant myeloid expression. In pediatric patients with the t(12;21)(ETV6-RUNX1) chimeric fusion transcript, aberrant expression of CD13 was observed in 39.13%, CD33 in 36.95%, and CD117 in 8.69% of patients, respectively. Pediatric BCP-ALL patients with the ETV6-RUNX1 fusion transcript and expression of aberrant myeloid markers were not significantly different compared to those without with respect to demographic and clinical/hematological characteristics (p=0.5955). Aberrant myeloid markers were rarely or never expressed in pediatric and adult BCP-ALL patients with the t(4;11)(KTM2A-AF4) and t(1;19)(TCF3-PBX1) fusion transcripts.

Conclusion: Aberrant myeloid markers were frequently expressed among BCP-ALL patients with the t(9;22)(BCR-ABL1) and t(12;21) (ETV6-RUNX1) fusion transcripts. However, BCP-ALL patients with the t(4;11)(KTM2A-AF4) and t(1;19)(TCF3-PBX1) fusion transcripts rarely or never expressed aberrant myeloid markers. Aberrant myeloid CD markers can be used in predicting chimeric fusion transcripts at baseline so as to plan appropriate tyrosine kinase inhibitor therapy in cases of BCP-ALL with specific chimeric fusion transcripts. This study has delineated the relationship of chimeric fusion transcripts with the aberrant expression of myeloid markers in a large cohort of BCP-ALL cases.

Keywords: Neoplasia; Acute leukemia; Acute lymphoblastic leukemias; Molecularbiology; Molecular hematology.

Conflict of interest statement

Conflict of Interest: No conflict of interest was declared by the authors.

Figures

Figure 1
Figure 1
Flow cytometric immunophenotyping of newly diagnosed adult B-cell precursor acute lymphoblastic leukemia (BCP-ALL) patients harboring the BCR-ABL1 fusion transcript with expression of aberrant myeloid markers. The blasts were gated on plots of CD45 versus side scatter (SS) (red populations) and were confirmed to be B-lineage blasts based on expression of B-lineage-associated markers (CD19, CD10, cytoplasmic CD79a). The gated blasts showed expression of myeloid-lineage associated markers (CD13 and CD33), which was considered aberrant expression of myeloid markers in cases of B-ALL.
Figure 2
Figure 2
Multiplex assay showing presence of minor transcript (e1a2, p190 kDa) and major transcripts (b2a2, b3a2, p210 kDa) of BCR-ABL1 in adult BCP-ALL cases. Lane 1 shows minor transcript-positive control/internal reference genes (e1a2) (521 bp), lane 7 shows positive minor transcript in a patient, lane 10 shows major transcript-positive control/internal reference genes (b2a2) (310 bp), lanes 12 and 13 show positive major transcripts (b2a2) in two patients, lane 15 shows major transcript-positive control/internal reference genes (b3a2) (385 bp), and lanes 17 and 18 show positive major transcripts (b3a2) in two patients.
Figure 3
Figure 3
Multiplex assay showing presence of t(4:11), t(1:19), and t(12:21) in adult BCP-ALL cases. Lane 1 of panel a shows t(4:11) (KMT2A-AF4)-positive control/internal reference genes (511 bp), lane 6 shows positive t(4:11)(KMT2A-AF4), and lane 3 shows t(1:19) (373 bp) in patients. Lane 1 of panel b shows t(12;21)(ETV6-RUNX1)-positive control/internal reference genes, lane 3 shows t(1:19)(TCF3-PBX1)-positive control/internal reference genes, and lane 4 shows t(12;21)(ETV6-RUNX1) (373 bp) transcripts.

References

    1. Dias A, Kenderian SJ, Westin GF, Litzow MR. Novel therapeutic strategies in acute lymphoblastic leukemia. Curr Hematol Malig Rep. 2016;11:253–264.
    1. Egler RA, Ahuja SP, Matloub Y. L-Asparaginase in the treatment of patients with acute lymphoblastic leukemia. J Pharmacol Pharmacother. 2016;7:62–71.
    1. Mullighan CG. The genomic landscape of acute lymphoblastic leukemia in children and young adults. Hematology Am Soc Hematol Educ Program. 2014;2014:174–180.
    1. Roberts KG, Li Y, Payne-Turner D, Harvey RC, Yang YL, Pei D, McCastlain K, Ding L, Lu C, Song G, Ma J, Becksfort J, Rusch M, Chen SC, Easton J, Cheng J, Boggs K, Santiago-Morales N, Iacobucci I, Fulton RS, Wen J, Valentine M, Cheng C, Paugh SW, Devidas M, Chen IM, Reshmi S, Smith A, Hedlund E, Gupta P, Nagahawatte P, Wu G, Chen X, Yergeau D, Vadodaria B, Mulder H, Winick NJ, Larsen EC, Carroll WL, Heerema NA, Carroll AJ, Grayson G, Tasian SK, Moore AS, Keller F, Frei-Jones M, Whitlock JA, Raetz EA, White DL, Hughes TP, Guidry Auvil JM, Smith MA, Marcucci G, Bloomfield CD, Mrózek K, Kohlschmidt J, Stock W, Kornblau SM, Konopleva M, Paietta E, Pui CH, Jeha S, Relling MV, Evans WE, Gerhard DS, Gastier-Foster JM, Mardis E, Wilson RK, Loh ML, Downing JR, Hunger SP, Willman CL, Zhang J, Mullighan CG. Targetable kinase-activating lesions in Ph-like acute lymphoblastic leukemia. N Engl J Med. 2014;371:1005–1015.
    1. Wang N, Huang L, Wang D, Wang J, Jiang L, Zhou K, Yang Y, Xu D, Zhou J. Successful engraftment of human acute lymphoblastic leukemia cells in NOD/SCID mice via intrasplenic inoculation. Cancer Biol Ther. 2012;13:1158–1164.
    1. Chiaretti S, Gianfelici V, O’Brien SM, Mullighan CG. Advances in the genetics and therapy of acute lymphoblastic leukemia. Am Soc Clin Oncol Educ Book. 2016;35:e314–322.
    1. Stock W. Adolescents and young adults with acute lymphoblastic leukemia. Hematology Am Soc Hematol Educ Program. 2010;2010:21–29.
    1. Bao L, Gross SA, Ryder J, Wang X, Ji M, Chen Y, Yang Y, Zhu S, Irons RD. Adult precursor B lymphoblastic leukemia in Shanghai, China: characterization of phenotype, cytogenetics and outcome for 137 consecutive cases. Int J Hematol. 2009;89:431–437.
    1. Bhatia P, Binota J, Varma N, Bansal D, Trehan A, Marwaha RK, Malhotra P, Varma S. Incidence of common chimeric fusion transcripts in B-cell acute lymphoblastic leukemia: an Indian perspective. Acta Haematol. 2012;128:17–19.
    1. Borkhardt A, Cazzaniga G, Viehmann S, Valsecchi MG, Ludwig WD, Burci L, Mangioni S, Schrappe M, Riehm H, Lampert F, Basso G, Masera G, Harbott J, Biondi A. Incidence and clinical relevance of TEL/AML1 fusion genes in children with acute lymphoblastic leukemia enrolled in the German and Italian multicenter therapy trials. Associazione Italiana Ematologia Oncologia Pediatrica and the Berlin-Frankfurt-Münster Study Group. Blood. 1997;15;90:571–577.
    1. Chopra A, Soni S, Verma D, Kumar D, Dwivedi R, Vishwanathan A, Vishwakama G, Bakhshi S, Seth R, Gogia A, Kumar L, Kumar R. Prevalence of common fusion transcripts in acute lymphoblastic leukemia: a report of 304 cases. Asia Pac J Clin Oncol. 2015;11:293–298.
    1. Chung HY, Kim KH, Jun KR, Jang S, Park CJ, Chi HS, Im HJ, Seo JJ, Seo EJ. Prognostic significance of TEL/AML1 rearrangement and its additional genetic changes in Korean childhood precursor B-acute lymphoblastic leukemia. Korean J Lab Med. 2010;30:1–8.
    1. Gupta N, Pawar R, Banerjee S, Brahma S, Rath A, Shewale S, Parihar M, Singh M, Arun SR, Krishnan S, Bhatacharyya A, Das A, Kumar J, Bhave S, Radhakrishnan V, Nair R, Chandy M, Arora N, Mishra D. Spectrum and immunophenotypic profile of acute leukemia: a tertiary center flow cytometry experience. Mediterr J Hematol Infect Dis. 2019;11:e2019017.
    1. Kerketta LS, Rao VB, Ghosh K. Chimeric fusion karyotypes in childhood B-cell acute lymphoblastic leukemia. Indian Pediatr. 2014;51:152–153.
    1. Liang DC, Shih LY, Yang CP, Hung IJ, Chen SH, Jaing TH, Liu HC, Chang WH. Multiplex RT-PCR assay for the detection of major fusion transcripts in Taiwanese children with B-lineage acute lymphoblastic leukemia. Med Pediatr Oncol. 2002;39:12–17.
    1. Martinez-Mancilla M, Rodriguez-Aguirre I, Tejocote-Romero I, Medina-Sanson A, Ocadiz-Delgado R, Gariglio P. Clinical relevance of the fusion transcripts distribution pattern in mexican children with acute lymphoblastic leukemia. J Pediatr Hematol Oncol. 2013;35:170–173.
    1. Olarte-Carrillo I, Ramos-Peñafiel CO, Miranda-Peralta E, Mendoza-Salas I, Nacho-Vargas KA, Zamora-Domínguez JA, Mendoza-García E, Rozen-Fuller E, Kassack-Ipiña JJ, Collazo-Jaloma J, Martínez-Tovar A. Frequency of the minor BCR-ABL (e1;a2) transcript oncogene in a Mexican population with adult acute lymphoblastic leukaemia. Revista Médica Del Hospital General De México. 2015;78:119–123.
    1. Sharma M, Sachdeva MU, Varma N, Varma S, Marwaha RK. Characterization of immunophenotypic aberrancies in adult and childhood acute lymphoblastic leukemia: a study from Northern India. J Cancer Res Ther. 2016;12:620–626.
    1. Siraj AK, Kamat S, Gutiérrez MI, Banavali S, Timpson G, Sazawal S, Bhargava M, Advani S, Adde M, Magrath I, Bhatia K. Frequencies of the major subgroups of precursor B-cell acute lymphoblastic leukemia in Indian children differ from the West. Leukemia. 2003;17:1192–1193.
    1. Sudhakar N, Rajalekshmy KR, Rajkumar T, Nancy KN. RT-PCR and real-time PCR analysis of E2A-PBX1, TEL-AML1, mBCR-ABL and KMT2A-AF4 fusion gene transcripts in de novo B-lineage acute lymphoblastic leukaemia patients in south India. J Genet. 2011;90:349–353.
    1. Trka J, Zuna J, Hrusák O, Kalinová M, Muzíková K, Lauschman H, Starý J. Impact of TEL/AML1-positive patients on age distribution of childhood acute lymphoblastic leukemia in Czech Republic. Pediatric Hematology Working Group in Czech Republic. Leukemia. 1998;12:996–997.
    1. Den Boer ML, Kapaun P, Pieters R, Kazemier KM, Janka-Schaub GE, Veerman AJ. Myeloid antigen co-expression in childhood acute lymphoblastic leukaemia: relationship with in vitro drug resistance. Br J Haematol. 1999;105:876–882.
    1. Gujral S, Badrinath Y, Kumar A, Subramanian PG, Raje G, Jain H, Pais A, Amre Kadam PS, Banavali SD, Arora B, Kumar P, Hari Menon VG, Kurkure PA, Parikh PM, Mahadik S, Chogule AB, Shinde SC, Nair CN. Immunophenotypic profile of acute leukemia: critical analysis and insights gained at a tertiary care center in India. Cytometry B Clin Cytom. 2009;76:199–205.
    1. Gupta M, Monga L, Mehrotra D, Chhabra S, Singhal S, Sen R. Immunophenotypic aberrancies in acute leukemia: a tertiary care centre experience. Oman Med J. 2021;36:e218.
    1. Seegmiller AC, Kroft SH, Karandikar NJ, McKenna RW. Characterization of immunophenotypic aberrancies in 200 cases of B acute lymphoblastic leukemia. Am J Clin Pathol. 2009;132:940–949.
    1. Supriyadi E, Veerman AJ, Purwanto I, van de Ven PM, Cloos J. Myeloid antigen expression in childhood acute lymphoblastic leukemia and its relevance for clinical outcome in Indonesian ALL-2006 Protocol. J Oncol. 2012;2012:135186.
    1. Borowitz MJ, Chan JK, Downing JR, Le Beau MM, Arber DA. B-Lymphoblastic leukaemia/lymphoma with recurrent genetic abnormalities. In: Swerdlow SH, Campo E, Harris NL, Jaffe ES, Pileri SA, Stein H, Thiele J (eds). WHO Classification of Tumours of Haematopoietic and Lymphoid Tissues. 4th Edition. Geneva. World Health Organization. 2017.
    1. Pakakasama S, Kajanachumpol S, Kanjanapongkul S, Sirachainan N, Meekaewkunchorn A, Ningsanond V, Hongeng S. Simple multiplex RT-PCR for identifying common fusion transcripts in childhood acute leukemia. Int J Lab Hematol. 2008;30:286–291.
    1. Raiya B, Vora H, Trivedi P, Parikh B. Flowcytometric analysis of leukemic blasts - as primary screening test for BCR/ABL1 gene rearrangement in B-ALL. EJMO. 2019;3:191–198.

Source: PubMed

Sponsors en medewerkers

Medische omstandigheden

Geneesmiddelinterventies

3
Abonneren