Mutational Analysis of EZH2 in the Germinal Centre Type of Diffuse Large B Cell Lymphoma

Authors

  • Farah Fatima Abbas
  • Muhammad Furqan Bari
  • Muhammad Nouman Mughal
  • Bilal Ahmed Khan
  • Nehad Khan
  • Saba Kamil
  • Abdul Razzaq
  • Noor Kamil

Keywords:

Germinal Center, Diffuse Large B Cell Lymphoma, Immunomarkers, EZH2 mutation, Polymerase chain reaction, Sequencing

Abstract

OBJECTIVE: To determine the Frequency of Germinal Center B (GCB) type Diffuse Large B Cell Lymphoma (DLBCL) in Karachi, Pakistan, and to screen the DLBCL patients for EZH2 mutation.
METHODOLOGY: A cross-sectional study was conducted on DLBCL patients, and IHC analysis, PCR, and sequencing of the EZH2 gene were used. This study was conducted from August 2016 to July 2020. Surgical specimens received in the histopathology section of DDRRL centrally and from different collection units throughout Sindh having been diagnosed with DLBCL were collected. Three markers (CD10, BCL6, and MUM1) were used for subtyping. DNA was manually extracted from the FFPE Blocks of DLBCL cases using phenol: chloroform: isopropanol alcohol. Polymerase chain reaction (PCR) was performed, and samples were sent from Dow University to Korea (Macrogen) for sequencing. The amplified product was 189bp. Primers were used for EZH2 to flank the region containing the tyrosine domain on exon 16 and exon 18. 
RESULTS: Out of 116 cases, 60.3% and 39.7% were male and female, respectively. The mean age was 49.73±16.38 years. The mean survival duration of cases diagnosed with GCB was 8.21±7.39 months. Polymorphism of EZH2 mutation (Tyr 701 Asn) was found to be positive in GCB-DLBCL for three samples (16.6%).
CONCLUSION: In the current study, Sanger sequencing was used to detect any mutation in the coding region of EZH2, and a unique mutation was reported (Tyr701Asn).

References

Piñeros M, Laversanne M, Barrios E, de Camargo Cancela M, de Vries E, Pardo C et al. An updated profile of the cancer burden, patterns and trends in Latin America and the Caribbean. Lancet Reg Health-Am. 2022 Sep; 13: 100294. doi: 10.1016/j.lana.2022.100294.

Mushtaq S, Akhtar N, Jamal S, Mamoon N, Khadim T, Sarfaraz T et al. Malignant lymphomas in Pakistan according to the WHO classification of lymphoid neoplasms. Asian Pac J Cancer Prev. 2008; 9(2): 229-32.

Lazim NM, Abdullah B. Multidisciplinary approach to children with sinonasal tumors: A review. Pediatr Investig. 2019; 3(3): 173-9. doi: 10.1002/ped4.12147.

Miranda-Filho A, Piñeros M, Znaor A, Marcos-Gragera R, Steliarova-Foucher E, Bray F. Global patterns and trends in the incidence of non-Hodgkin lymphoma. Cancer Causes Control. 2019; 30(5): 489-99. doi: 10.1007/s10552-019-01155-5.

Khorshid O, Salama A, Helal A, Sallam Y, Amin M. Clinicopathologic profile and outcome of extranodal diffuse large B-cell NHL Egyptian National Cancer Institute Experience. Forum Clin Oncol. 2015; 6(3): 8-12. doi: 10.1515/fco-2015-0013.

Menon MP, Pittaluga S, Jaffe ES. The histological and biological spectrum of diffuse large B-cell lymphoma in the World Health Organization classification. Cancer J. 2012; 18(5): 411-420. doi: 10.1097/PPO.0b013e31826aee97.

Alyahya N, Adiga B, Alwadei A, Alshahrani G, Alyahya F. The clinicopathological profile of non-Hodgkin's lymphoma in Aseer region of Saudi Arabia. BMC Res Notes. 2019; 12(1): 418. doi: 10.1186/s13104-019-4447-1.

Diumenjo MC, Abriata G, Forman D, Sierra MS. The burden of non-Hodgkin lymphoma in Central and South America. Cancer Epidemiol. 2016; 44: S168-S177. doi: 10.1016/j.canep. 2016.05.008.

Mahmood H, Habib M, Aslam W, Khursheed S, Fatima S, Aziz S et al. Clinicopathological spectrum of Diffuse Large B Cell lymphoma: a study targeting population yet unexplored in Pakistan. BMC Res Notes. 2021; 14: 1-6. doi: 10.1186/s13104-021-05768-5.

Ali S, Ali M, Badar F, Basit A, Hameed A. Factors Associated with Increased Red Blood Cells Transfusion Requirements in Patients with Hodgkin and Non-Hodgkin Lymphoma. J Ayub Med Coll Abbottabad. 2015; 27(1): 70-3.

Padala SA, Kallam A. Diffuse large B cell lymphoma. In: StatPearls [Internet]. Treasure Island (FL): StatPearls Publishing; 2024 Jan, 2023 Apr 24.

Johnson N, Assouline S, Alcaide M, Mohajeri A, Greenwood C, Riazalhosseini Y et al. Genetic landscapes of relapsed and refractory diffuse large B cell lymphomas. Cancer Res. 2016; 76(14):117. doi: 10.1158/1538-7445.AM2016-117.

Colomo L, López-Guillermo A, Perales M, Rives S, Martínez A, Bosch F et al. Clinical impact of the differentiation profile assessed by immunophenotyping in patients with diffuse large B-cell lymphoma. Blood. 2003; 101(1): 78-84. doi: 10.1182/blood-2002-04-1286. Epub 2002 Aug 8.

Chapman JR, Bouska AC, Zhang W, Alderuccio JP, Lossos IS, Rimsza LM et al. EBV?positive HIV?associated diffuse large B cell lymphomas are characterized by JAK/STAT (STAT3) pathway mutations and unique clinicopathologic features. Br J Haematol. 2021; 194(5): 870-878. doi: 10.1111/bjh.17708. Epub 2021 Jul 17.

Li SS, Zhai XH, Liu HL, Liu TZ, Cao TY, Chen DM et al. Whole-exome sequencing analysis identifies distinct mutational profile and novel prognostic biomarkers in primary gastrointestinal diffuse large B-cell lymphoma. Exp Hematol Oncol. 2022; 11(1): 71. doi: 10.1186/s40164-022-00325-7.

Narazaki T, Nakashima Y, Tsukamoto Y, Tsuda M, Masuda T, Kimura D et al. Transformation of follicular lymphoma to double-hit lymphoma during adjuvant chemotherapy for concurrent ovarian carcinoma. Int J Hematol. 2019; 110(3): 375-80. doi: 10.1007/s12185-019-02656-4. Epub 2019 May 18.

Erokhin M, Chetverina O, Gy?rffy B, Tatarskiy VV, Mogila V, Shtil AA et al. Clinical correlations of polycomb repressive complex 2 in different tumor types. Cancers(Basel). 2021; 13(13): 3155. doi: 10.3390/cancers13133155.

Li B, Chng WJ. EZH2 abnormalities in lymphoid malignancies: underlying mechanisms and therapeutic implications. J Hematol Oncol. 2019; 12(1): 118. doi: 10.1186/s13045-019-0814-6.

Dharanipragada P, Parekh N. Genome-wide characterization of copy number variations in diffuse large B-cell lymphoma with implications in targeted therapy. Precis Clin Med. 2019; 2(4): 246-58. doi: 10.1093/pcmedi/pbz024. Epub 2019 Nov 21.

Morin R, Bainbridge M, Fejes A, Hirst M, Krzywinski M, Pugh T et al. Profiling the HeLa S3 transcriptome using randomly primed cDNA and massively parallel short-read sequencing. Biotechniques. 2008; 45(1): 81-94. doi: 10.2144/000112900.

Chen G, Shi T, Shi L. Characterizing and annotating the genome using RNA-seq data. Sci China Life Sci. 2017; 60(2): 116-125. doi: 10.1007/s11427-015-0349-4. Epub 2016 Jun 13.

Montesinos-Rongen M, Brunn A, Sanchez-Ruiz M, Küppers R, Siebert R, Deckert M. Impact of a Faulty Germinal Center Reaction on the Pathogenesis of Primary Diffuse Large B Cell Lymphoma of the Central Nervous System. Cancers(Basel). 2021; 13(24): 6334. doi: 10.3390/cancers13246334.

Sneeringer CJ, Scott MP, Kuntz KW, Knutson SK, Pollock RM, Richon VM et al. Coordinated activities of wild-type plus mutant EZH2 drive tumor-associated hypertrimethylation of lysine 27 on histone H3 (H3K27) in human B-cell lymphomas. Proc Natl Acad Sci U S A. 2010; 107(49): 20980-5. doi: 10.1073/pnas.1012525107. Epub 2010 Nov 15.

Lee EF, Fairlie WD. Discovery, development and application of drugs targeting BCL-2 pro-survival proteins in cancer. Biochem Soc Trans. 2021; 49(5): 2381-95. doi: 10.1042/BST20210749.

Merdan S, Subramanian K, Ayer T, Van Weyenbergh J, Chang A, Koff JL et al. Gene expression profiling-based risk prediction and profiles of immune infiltration in diffuse large B-cell lymphoma. Blood Cancer J. 2021; 11(1): 2. doi: 10.1038/s41408-020-00404-0.

Zamò A, Gerhard-Hartmann E, Ott G, Anagnostopoulos I, Scott DW, Rosenwald A et al. Routine application of the Lymph2Cx assay for the subclassification of aggressive B-cell lymphoma: report of a prospective real-world series. Virchows Arch. 2022; 481(6): 935-943. doi: 10.1007/s00428-022-03420-6. Epub 2022 Oct 11.

Downloads

Published

2024-04-03

How to Cite

1.
Abbas FF, Bari MF, Mughal MN, Khan BA, Khan N, Kamil S, Razzaq A, Kamil N. Mutational Analysis of EZH2 in the Germinal Centre Type of Diffuse Large B Cell Lymphoma. J Liaq Uni Med Health Sci [Internet]. 2024 Apr. 3 [cited 2024 Apr. 22];23(01):15-20. Available from: http://ojs.lumhs.edu.pk/index.php/jlumhs/article/view/1173