Gene-edited MLE-15 Cells as a Model for the Hermansky-Pudlak Syndromes.

Kook S, Qi A, Wang P, Meng S, Gulleman P, Young LR, Guttentag SH
Am J Respir Cell Mol Biol. 2018 58 (5): 566-574

PMID: 29190429 · PMCID: PMC5946333 · DOI:10.1165/rcmb.2017-0324MA

Defining the mechanisms of cellular pathogenesis in rare lung diseases such as Hermansky-Pudlak syndrome (HPS) is often complicated by loss of the differentiated phenotype of cultured primary alveolar type 2 (AT2) cells, as well as by a lack of durable cell lines that are faithful to both AT2-cell and rare disease phenotypes. We used CRISPR/Cas9 gene editing to generate a series of HPS-specific mutations in the MLE-15 cell line. The resulting MLE-15/HPS cell lines exhibit preservation of AT2 cellular functions, including formation of lamellar body-like organelles, complete processing of surfactant protein B, and known features of HPS specific to each trafficking complex, including loss of protein targeting to lamellar bodies. MLE-15/HPS1 and MLE-15/HPS2 (with a mutation in Ap3β1) express increased macrophage chemotactic protein-1, a well-described mediator of alveolitis in patients with HPS and in mouse models. We show that MLE-15/HPS9 and pallid AT2 cells (with a mutation in Bloc1s6) also express increased macrophage chemotactic protein-1, suggesting that mice and humans with BLOC-1 mutations may also be susceptible to alveolitis. In addition to providing a flexible platform to examine the role of HPS-specific mutations in trafficking AT2 cells, MLE-15/HPS cell lines provide a durable resource for high-throughput screening and studies of cellular pathophysiology that are likely to accelerate progress toward developing novel therapies for this rare lung disease.

MeSH Terms (16)

Alveolar Epithelial Cells Animals Cell Line Clustered Regularly Interspaced Short Palindromic Repeats CRISPR-Associated Protein 9 CRISPR-Cas Systems Disease Models, Animal Gene Editing Genetic Markers Genetic Predisposition to Disease Hermanski-Pudlak Syndrome Humans Mice, Inbred C57BL Mice, Transgenic Mutation Phenotype

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