Summary
B-cell non-Hodgkin’s lymphomas are a haematologically, genetically, and clinically heterogeneous group of neoplasms, with most emerging from B lymphocytes in the germinal centre. It accounts for approximately 90% of all non-Hodgkin’s lymphomas. Diffuse large B-cell lymphomas, follicular lymphoma, Burkitt lymphoma, and B-cell chronic lymphocytic leukaemia/small lymphocytic lymphoma are typical B-NHL subtypes.
The current study investigated the haematological and biochemical observations and identified the molecular and genomic signature of B-cell non Hodgkin Lymphoma in Kurdistanian patients using the most recent techniques including Whole transcriptome next generation sequencing NGS and quantitative real time polymerase chain reaction (q-RT PCR).
The study aimed to investigate the haematological and biochemical evaluations supported by more specific potential molecular biomarkers to quantify the diagnostic and prognostic value of colony stimulating factor-1 receptor (CSF1R), myeloid differentiation factor 88 (MYD88) and tumor necrosis factor-α (TNF-α) oncogenes, and to provide essential insights for detection of polymorphisms and novel mutations in B-NHL.
The current study involving 73 subjects of controls and NHL in the Kurdistan/Iraq, were recruited from Nanakali Hospital for Blood Diseases and Cancer-Erbil- Iraq. The samples of blood and paraffin-embedded tissue were selected from individuals which are not treated and newly diagnosed non-Hodgkin lymphoma (NHL) and normal tissue specimens, including 49 non-Hodgkin lymphoma patients (22 females and 27 males) and 24 controls (12 females and 12 males). The samples stratified by gender and age.
This study evaluated some aspects of NHL divided into three study parts. The first one is the assessment of haematological and biochemical variables in for 73 NHL and control groups. The second is the identification of genomic variations and mutations done for eight B-NHL patients uses next generation sequencing for target genes, CSF1R, MYD88 and the TNF α in NHL. The third part is the quantification of gene expression levels via quantitative real time polymerase chain reaction (q-RT PCR) technique for 73 samples, focusing on three key genes MYD88, TNF, and CSF1R in which expression profiles analysed in B-NHL patients and controls.
The Hematological laboratory parameters measured are red blood cell (RBC) count, hemoglobin (Hb), hematocrit (Hct), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), red blood cell distribution width (RDW), white blood cells (WBCs) count, differential leukocyte count, neutrophils, monocytes (MON), eosinophils (EOS), lymphocytes (LYM), platelet (PLT) count, mean platelet volume (MPV) were measured by Beckman Coulter. The serum biochemical parameters were determined such as aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), lactate dehydrogenase (LDH), C-reactive protein (CRP), creatinine, urea, uric acid, total bilirubin and total protein by Cobas c311.
The study results had a more incidence of NHL in males than in females and more frequent in > 50 years than ≤ 50 years NHL patients. A greater number of cases were categorized as DLBCL and the most common was stage IV of NHL cases. The initial results of the hematological parameters regarding gender and age in NHL patients exhibited highly significant reductions in RBC counts, hemoglobin, haematocrit and mean corpuscular volume, while significant increase in red distribution width compared to control groups in both sex and all age groups. Also, white blood cell counts show significant increase in males NHL patients >50 years compared to control group. The biochemical findings showed significantly higher c-reactive protein, lactate dehydrogenase and alkaline phosphatase in NHL patients in both gender and age groups, reduction in total protein and albumin for females and males of ≤50 years NHL patients, increase in urea levels for females ≤50 years and males >50 years and uric acid for NHL patients of both gender and age groups compared to control groups, reduction in creatinine for both gender and calcium in males ≤50 years NHL patients compared with control groups. The molecular outcomes present the detection of novel variations and different mutated genes for the three target genes, especially, for the CSF1R gene. Furthermore, the findings show the overall lower CSF1R expression in B-NHL as compared to the controls and significant reduction in CSF1R expression in females (≤50 years and >50 years). The result considers lower CSF1R expression in B-NHL males (≤50 years), and higher but not significant in males (>50 years). Hence, the outcomes show specific differences in gene expression regarding to the gender for the three target genes, especially for CSF1R.
These B-NHL genetic mutations and levels of expressed genes may be considered as potential diagnostic markers with their meaningful comparisons to control groups; they could be proposed to guide the management of patients and facilitate their stratification into clinical trials. Also, this study found the significance of gene mutations and gene expression levels for CSF1R, MYD88 and TNF-α as prognostic and diagnostic biomarkers in NHL.