The current study has been designed to determine the frequency of CEBPA gene mutations in cohort of 30 acute myeloid leukemia (AML) patients, and their role in the development of AML, and their effect on the structure and function of translated protein was also studied. Blood samples were collected from 30 patients from Jinnah and Mayo hospitals, and their hematological characteristics were noted. DNA was isolated from all blood samples, and after amplification, samples were sent for sequencing. Mutations were identified by comparing sequencing results with normal CEBPA gene nucleotide sequences. CEBPA gene mutations were observed in 7 (23.3%) 30 patients. These 7 patients had two missense mutations both in coding and non-coding regions. Non-coding region mutation was detected in 4 patients, while coding region mutation was observed in 3 patients. There was no significant difference in hemoglobin level, TLC, platelets count, and blast count percentage between CEBPA gene mutant and non-mutant AML patients. While high LDH was observed in CEBPA mutant patients. The deleterious effect of S348T mutation was checked using SNAP2, PROVEAN, PolyPhen-2, PhD-SNP, Align GVGD, and PANTHER. Protein's stability was studied by using I-Mutant 3.0. ConSurf 2.0 tools were used to check the conservation status of mutation, while NetsurfP predicted protein's secondary structure. The SWISS model generated a 3D model of both mutant and wild-type residues, which MolProbity, Protparam, ProSA web, and VADAR analysis further analyzed. The mutational effect on RNA's secondary structure was analyzed using the Vienna package, while the interaction of CEBPA gene with other proteins was analyzed by STRING tool.

Keywords:  myeloid leukemia, mutations,