Hereditary neutropenia is usually caused by heterozygous germline mutations in the ELANE gene encoding neutrophil elastase (NE). How mutations cause disease remains uncertain, but two hypotheses have been proposed. In one, ELANE mutations lead to mislocalization of NE. In the other, ELANE mutations disturb protein folding, inducing an unfolded protein response in the endoplasmic reticulum (ER). In this study, we describe new types of mutations that disrupt the translational start site. At first glance, they should block translation and are incompatible with either the mislocalization or misfolding hypotheses, which require mutant protein for pathogenicity. We find that start-site mutations, instead, force translation from downstream in-frame initiation codons, yielding amino-terminally truncated isoforms lacking ER-localizing (pre) and zymogen-maintaining (pro) sequences, yet retain essential catalytic residues. Patient-derived inducedpluripotent stemcells recapitulate hematopoietic andmolecular phenotypes. Expression of theaminoterminally deleted isoforms in vitro reducesmyeloid cell clonogenic capacity.We define an internal ribosome entry site (IRES)within ELANE and demonstrate that adjacentmutations modulate IRES activity, independently of protein-coding sequence alterations. Some ELANE mutations, therefore, appear to cause neutropenia via the production of amino-terminally deleted NE isoforms rather than by altering the coding sequence of the full-length protein. © 2014 by The American Society of Hematology.
Tidwell, T., Wechsler, J., Nayak, R. C., Trump, L., Salipante, S. J., Cheng, J. C., … Horwitz, M. S. (2014). Neutropenia-associated ELANE mutations disrupting translation initiation produce novel neutrophil elastase isoforms. Blood, 123(4), 562–569. https://doi.org/10.1182/blood-2013-07-513242