Abstract
Mutations in the Natural Killer Group 2D (NKG2D) gene are associated with an elevated risk of many malignancies, including Chronic Myeloid leukemia (CML). The mutational landscape of exon 8 of the NKG2D gene is poorly defined, especially in CML. Therefore, this study aims to investigate mutations in exon 8 of the NKG2D gene in CML patients, analyze its potential interactions with other genes, and assess related biochemical, hematological, and clinicopathological parameters.
Blood samples were obtained from a total of 50 participants, including 25 patients diagnosed with chronic myeloid leukemia (CML) and 25 healthy controls, for molecular, biochemical, and hematological analyses. Molecular analyses included confirmation of BCR-ABL positivity in CML patients, DNA extraction, polymerase chain reaction (PCR) amplification of exon 8, gel electrophoresis, and Sanger sequencing. In addition, biochemical and hematological parameters were analyzed for both CML patients and healthy participants.
Sanger sequencing was used to identify variations in exon 8 of the NKG2D gene, and the GeneMANIA database was used to predict gene interactions. The result showed that exon 8 alterations (substitution or deletion) were found in 88% of CML patients, with 50% exhibiting multiple variations. Nine single nucleotide polymorphisms (SNPs) were found, including eight substitutions and one deletion. The most common substitution was G>GC and six variants were unique. The homozygous 414G>C variation had the highest mutation score.
The GeneMANIA study revealed strong interactions between NKG2D and 20 other genes, particularly the HCST gene, with notable co-expression and pathway linkages. Biochemically, serum Lactate dehydrogenase (LDH) levels were considerably greater in CML patients, while uric acid levels were not significantly different. The MID%, representing mid-sized cells such as eosinophils, and basophils, was higher in CML patients, whereas red blood cell (RBC) count, hemoglobin, and hematocrit were lower than in controls. To summarize, exon 8 mutations of the NKG2D gene are quite common among CML patients, with numerous and unique variants. These mutations, together with NKG2D’s robust interaction network, may play a significant role in leukemogenesis and immunological dysregulation in CML.
