Lymphocyte cytosolic protein 1 (L-plastin) I232F mutation impairs granulocytic proliferation and causes neutropenia

• Published in: Blood advances 2022-04, Vol.6 (8), p.2581-2594
• Main Authors: Mahat, Upendra, Garg, Bhavuk, Yang, Chao-Yie, Mehta, Hrishikesh, Hanna, Rabi, Rogers, Heesun J., Flagg, Aron, Ivanov, Andrei I., Corey, Seth J.
• Format: Article
• Language: English
• Subjects: Actins - genetics; Cell Proliferation; Child; HeLa Cells; Humans; Lymphocytes; Membrane Glycoproteins; Microfilament Proteins; Mutation; Neutropenia - genetics; Phagocytes, Granulocytes, and Myelopoiesis
• ISSN: 2473-9529; 2473-9537
• DOI: 10.1182/bloodadvances.2021006398

Neutrophils migrate into inflamed tissue, engage in phagocytosis, and clear pathogens or apoptotic cells. These processes require well-coordinated events involving the actin cytoskeleton. We describe a child with severe neutropenia and episodes of soft tissue infections and pneumonia. Bone marrow examination showed granulocytic hypoplasia with dysplasia. Whole-exome sequencing revealed a de novo heterozygous missense mutation in LCP1, which encodes the F-actin–binding protein Lymphocyte Cytosolic Protein 1. To determine its pathophysiological significance, we stably transduced cells with doxycycline-inducible wild-type LCP1 and LCP1 I232F lentiviral constructs. We observed dysplastic granulocytic 32D cells expressing LCP1 I232F cells. These cells showed decreased proliferation without a block in differentiation. In addition, expression of LCP1 I232F resulted in a cell cycle arrest at the G2/M phase, but it did not lead to increased levels of genes involved in apoptosis or the unfolded protein response. Both 32D and HeLa cells expressing mutant LCP1 displayed impaired cell motility and invasiveness. Flow cytometry showed increased F-actin. However, mutant LCP1-expressing 32D cells exhibited normal oxidative burst upon stimulation. Confocal imaging and subcellular fractionation revealed diffuse intracellular localization of LCP1, but only the mutant form was found in the nucleus. We conclude that LCP1 is a new gene involved in granulopoiesis, and the missense variant LCP1 I232F leads to neutropenia and granulocytic dysplasia with aberrant actin dynamics. Our work supports a model of neutropenia due to aberrant actin regulation. •A missense mutation in LCP1, an actin-bundling protein, results in severe neutropenia, migration defects, and susceptibility to infection.•LCP1 mutation causes actin dysregulation with increased F-actin, G2/M cell cycle arrest, dysplastic cells, and nuclear mislocalization. [Display omitted]