Expression properties, structural features and functional analysis of CALR E381A in MPN patients.

OBJECTIVE

To investigate the expression properties, structural features and function of CALR E381A in myeloproliferative neoplasms (MPN) patients.

METHODS

In this retrospective study, 435 MPN patients admitted to the Department of Hematology, Ningbo First Hospital from July 2015 to July 2021 were selected as the study subjects. Mutations in exon 9 from genomic DNA samples were identified by PCR, followed by Sanger sequencing. The physicochemical properties of the wild-type calreticulin and the p.E381A variant, and the structural information of the p.E381A variant were analyzed by using the bioinformatics databases. Growth assay of UT-7/mpl cells with E381A was used for the functional analysis of CALR E381A.

RESULTS

The predominant types of variants were identified as follows: p.L367fs46 (38.1%), p.K385fs47 (25.8%) and p.E381A (19.6%). Notably, the frequency of the p.E381A variant (c.1142A >C) in polycythemia vera or essential thrombocythemia was significantly higher than the frequency of that as a single nucleotide polymorphism (SNP) in the East Asian population. Furthermore, E381A coexisted with other genetic variants, of which V617F was more common. Bioinformatics analysis confirmed that E381A did not change the physicochemical properties of the calreticulin protein, but did change the electrical charge, energy state and steric hindrance of amino acid residues at site 381. UT-7/mpl cells harboring E381A overexpression did not exhibit altered cell growth, which is distinctly different from the stereotypical frameshift mutation.

CONCLUSION

CALR E381A is not a driver mutation for the development of MPN but may be a risk SNP implying an inherited predisposition for MPN disease in East Asian populations.