IGSF3 Requirement for Repair of Human Lung Endothelial Cell Injury

作者：C. Moeder, Monica J. Justice, Leonardo Jost, B. Dubois, S. Mousavi Aghdas, T. Rivera, Joanna M. Poczobutt, I. Petrache, Max S. Hiltabidle · 发表于：American Journal of Respiratory and Critical Care Medicine · 年份：2025 · DOI：10.1164/ajrccm.2025.211.abstracts.a4636 · 研究领域：Pulmonary Hypertension Research and Treatments

Abstract Rationale: By altering cell migration, the tetraspannin interacting protein Immunoglobulin superfamily 3 (IGSF3) impairs lung epithelial wound repair and barrier function during cigarette smoke exposure (Schweitzer et al., JCI insight 2020) and contributes to cancer metastases (Guo et al., Cancer sci. 2024). Although both known IGSF3 isoforms (IGSF3-1 and IGSF3-2) are expressed in lung endothelial cells, their role in lung endothelial cell function is less defined. We hypothesized that IGSF3 is required for endothelial cell migration and barrier function in both homeostatic conditions and during injury. Methods: Gain- and loss of function of IGSF3 in transformed human lung microvascular endothelial cells (HULEC) were achieved via lentivirus transduction of IGSF3 overexpressing stable cell lines (isoforms 1 and 2, each C-terminally or N-terminally V5-tagged) and via targeting CRISPR-Cas9, respectively. Electric Cell-substrate Impedance sensing (ECIS) was used for measuring cell proliferation rates (transcellular capacitance) as well as monolayer barrier function and repair (trans-endothelial cell resistance, TER) following electrical heat-induced wounding and electrical fencing. Cell proliferation was measured by CCK8 assay. Results: After confirming overexpression by western blotting, trans-endothelial capacitance measurements for 24 hours indicated that overexpression of both IGSF3 isoforms significantly decreased cellular proliferation, from 1.3-fold (N-tagged isof...