Harnessing human iPSC-microglia for CNS-wide delivery of disease-modifying proteins

Jean Paul Chadarevian¹, Hayk Davtyan², Alina L Chadarevian¹, Jasmine Nguyen³, Joia K Capocchi³, Lauren Le³, Adrian Escobar⁴, Talar Chadarevian⁴, Kimiya Mansour⁴, Ekaterina Deynega³, Michael Mgerian⁴, Christina Tu², Sepideh Kiani Shabestari¹, William Carlen-Jones³, Ghazaleh Eskandari-Sedighi³, Jonathan Hasselmann², Robert C Spitale⁵, Mathew Blurton-Jones⁶

Affiliations

¹ Department of Neurobiology & Behavior, University of California, Irvine, Irvine, CA 92697, USA; Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
² Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
³ Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA.
⁴ Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
⁵ Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, CA 92697, USA. Electronic address: rspitale@uci.edu.
⁶ Department of Neurobiology & Behavior, University of California, Irvine, Irvine, CA 92697, USA; Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA. Electronic address: mblurton@uci.edu.

PMID: 40233761
PMCID: PMC12360426 (available on 2026-04-14)
DOI: 10.1016/j.stem.2025.03.009

Harnessing human iPSC-microglia for CNS-wide delivery of disease-modifying proteins

Jean Paul Chadarevian et al. Cell Stem Cell. 2025.

. 2025 Jun 5;32(6):914-934.e8.

doi: 10.1016/j.stem.2025.03.009. Epub 2025 Apr 14.

Authors

Affiliations

¹ Department of Neurobiology & Behavior, University of California, Irvine, Irvine, CA 92697, USA; Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
² Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
³ Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA.
⁴ Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
⁵ Department of Pharmaceutical Sciences, University of California, Irvine, Irvine, CA 92697, USA. Electronic address: rspitale@uci.edu.
⁶ Department of Neurobiology & Behavior, University of California, Irvine, Irvine, CA 92697, USA; Institute for Memory Impairments and Neurological Disorders, University of California, Irvine, Irvine, CA 92697, USA; Sue and Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA. Electronic address: mblurton@uci.edu.

PMID: 40233761
PMCID: PMC12360426 (available on 2026-04-14)
DOI: 10.1016/j.stem.2025.03.009

Abstract

Widespread delivery of therapeutic proteins to the brain remains challenging. To determine whether human induced pluripotent stem cell (iPSC)-microglia (iMG) could enable brain-wide and pathology-responsive delivery of therapeutic cargo, we utilized CRISPR gene editing to engineer iMG to express the Aβ-degrading enzyme neprilysin under control of the plaque-responsive promoter, CD9. To further determine whether increased engraftment enhances efficacy, we utilized a CSF1R-inhibitor resistance approach. Interestingly, both localized and brain-wide engraftment in Alzheimer's disease (AD) mice reduced multiple biochemical measures of pathology. However, within the plaque-dense subiculum, reductions in plaque load, dystrophic neurites, and astrogliosis and preservation of neuronal density were only achieved following widespread microglial engraftment. Lastly, we examined chimeric models of breast cancer brain metastases and demyelination, demonstrating that iMG adopt diverse transcriptional responses to differing neuropathologies, which could be harnessed to enable widespread and pathology-responsive delivery of therapeutics to the CNS.

Keywords: Alzheimer’s disease; CRISPR; iMG; iPSC; immune cell therapy; microglia; microglia replacement; neprilysin; neurodegeneration; payload delivery.

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Conflict of interest statement

Declaration of interests J.P.C., H.D., J.H., R.C.S., and M.B.-J. are co-inventors on patent applications filed by the University of California Regents related to genetic modification of cells to confer resistance to CSF1R antagonists (WO2022/212897), transplantation of stem-cell-derived microglia to treat leukodystrophies (WO2024/015933), and pathology-responsive recombinant cells (WO2023/212663). M.B.-J. is a co-inventor of patent application related to the differentiation of human pluripotent stem cells into microglia (WO2018/160496). M.B.-J. is a co-founder and consultant for Savanna Biotherapeutics (formally NovoGlia Inc.). R.C.S. is a co-founder and board member of Savanna Biotherapeutics.

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Harnessing human iPSC-microglia for CNS-wide delivery of disease-modifying proteins

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Harnessing human iPSC-microglia for CNS-wide delivery of disease-modifying proteins

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Conflict of interest statement

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