Engineering an inhibitor-resistant human CSF1R variant for microglia replacement.

Hematopoietic stem cell transplantation (HSCT) can replace endogenous microglia with circulation-derived macrophages but has high mortality. To mitigate the risks of HSCT and expand the potential for microglia replacement, we engineered an inhibitor-resistant CSF1R that enables robust microglia replacement. A glycine to alanine substitution at position 795 of human CSF1R (G795A) confers resistance to multiple CSF1R inhibitors, including PLX3397 and PLX5622. Biochemical and cell-based assays show no discernable gain or loss of function. G795A- but not wildtype-CSF1R expressing macrophages efficiently engraft the brain of PLX3397-treated mice and persist after cessation of inhibitor treatment. To gauge translational potential, we CRISPR engineered human-induced pluripotent stem cell-derived microglia (iMG) to express G795A. Xenotransplantation studies demonstrate that G795A-iMG exhibit nearly identical gene expression to wildtype iMG, respond to inflammatory stimuli, and progressively expand in the presence of PLX3397, replacing endogenous microglia to fully occupy the brain. In sum, we engineered a human CSF1R variant that enables nontoxic, cell type, and tissue-specific replacement of microglia. © 2022 Chadarevian et al.

Citation

Jean Paul Chadarevian, Sonia I Lombroso, Graham C Peet, Jonathan Hasselmann, Christina Tu, Dave E Marzan, Joia Capocchi, Freddy S Purnell, Kelsey M Nemec, Alina Lahian, Adrian Escobar, Whitney England, Sai Chaluvadi, Carleigh A O'Brien, Fazeela Yaqoob, William H Aisenberg, Matias Porras-Paniagua, Mariko L Bennett, Hayk Davtyan, Robert C Spitale, Mathew Blurton-Jones, F Chris Bennett. Engineering an inhibitor-resistant human CSF1R variant for microglia replacement. The Journal of experimental medicine. 2023 Mar 06;220(3)

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PMID: 36584406

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