Direct microglia replacement reveals pathologic and therapeutic contributions of brain macrophages to a monogenic neurological disease

William H Aisenberg¹, Carleigh A O'Brien¹, Madison Sangster², Fazeela Yaqoob¹, Yuanchao Zhang³, Brian Temsamrit⁴, Searlait Thom⁵, Luca Gosse⁵, Sai Chaluvadi¹, Bilal Elfayomi¹, Gavin Lee¹, Vidhur Polam¹, Eli M Levitt¹, Gary Liu¹, Sonia I Lombroso¹, Kelsey M Nemec¹, Gavin Clowry⁶, Cassaundra Nieves², Priyanka Rawat², Emily Church¹, Daniel Martinez⁷, Clarissa Shoffler⁸, Daliya Kancheva⁹, Christopher Petucci⁸, Deanne Taylor¹⁰, Julia Kofler¹¹, Daniel Erskine⁵, Kiavash Movahedi⁹, Mariko L Bennett¹², F Chris Bennett¹³

Affiliations

¹ Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
² Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
³ Department of Biomedical Informatics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
⁴ Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
⁵ Translational and Clinical Research Institute, Newcastle University, Newcastle upon Tyne, UK.
⁶ Biosciences Institute, Newcastle University, Newcastle upon Tyne, UK.
⁷ Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
⁸ Penn Metabolomics Core, Cardiovascular Institute, Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
⁹ Brain and Systems Immunology Laboratory, Brussels Center for Immunology, Vrije Universiteit Brussel, Brussels, Belgium.
¹⁰ Department of Biomedical Informatics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA; Department of Pediatrics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
¹¹ Division of Neuropathology, Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA.
¹² Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA; Department of Pediatrics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Department of Neurology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
¹³ Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA. Electronic address: frederick.bennett@pennmedicine.upenn.edu.

PMID: 40311614
PMCID: PMC12078009 (available on 2026-05-13)
DOI: 10.1016/j.immuni.2025.03.019

Direct microglia replacement reveals pathologic and therapeutic contributions of brain macrophages to a monogenic neurological disease

William H Aisenberg et al. Immunity. 2025.

. 2025 May 13;58(5):1254-1268.e9.

doi: 10.1016/j.immuni.2025.03.019. Epub 2025 Apr 30.

Authors

Affiliations

¹ Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
² Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
³ Department of Biomedical Informatics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
⁴ Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA.
⁵ Translational and Clinical Research Institute, Newcastle University, Newcastle upon Tyne, UK.
⁶ Biosciences Institute, Newcastle University, Newcastle upon Tyne, UK.
⁷ Department of Pathology and Laboratory Medicine, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA.
⁸ Penn Metabolomics Core, Cardiovascular Institute, Department of Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA 19104, USA.
⁹ Brain and Systems Immunology Laboratory, Brussels Center for Immunology, Vrije Universiteit Brussel, Brussels, Belgium.
¹⁰ Department of Biomedical Informatics, Children's Hospital of Philadelphia, Philadelphia, PA 19104, USA; Department of Pediatrics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
¹¹ Division of Neuropathology, Department of Pathology, University of Pittsburgh, Pittsburgh, PA, USA.
¹² Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA; Department of Pediatrics, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA; Department of Neurology, University of Pennsylvania Perelman School of Medicine, Philadelphia, PA 19104, USA.
¹³ Department of Psychiatry, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA; Division of Neurology, Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, PA, USA. Electronic address: frederick.bennett@pennmedicine.upenn.edu.

PMID: 40311614
PMCID: PMC12078009 (available on 2026-05-13)
DOI: 10.1016/j.immuni.2025.03.019

Abstract

Krabbe disease, also named globoid cell (GC) leukodystrophy (GLD) for its distinct lipid-laden macrophages, is a severe leukodystrophy caused by galactosylceramidase (GALC) mutations. Hematopoietic stem cell transplant (HSCT) ameliorates disease and is associated with central nervous system (CNS) engraftment of GALC⁺ donor macrophages. Yet, the role of macrophages in GLD pathophysiology and HSCT remains unclear. Using single-cell sequencing, we revealed early interferon response signatures that preceded progressively severe macrophage dyshomeostasis and identified a molecular signature of GCs, which we validated in human brain specimens. Genetic depletion and direct microglia replacement by CNS monocyte injection rapidly replaced >80% of endogenous microglia with healthy macrophages in the twitcher (Galc^W355^∗) mouse model of GLD. Perinatal microglia replacement completely normalized transcriptional signatures, rescued histopathology, and doubled average survival. Overall, we uncovered distinct forms of microglial dysfunction and evidence that direct, CNS-limited microglia replacement improves a monogenic neurodegenerative disease, identifying a promising therapeutic target.

Keywords: Krabbe disease; disease-associated macrophage; globoid cell; globoid cell leukodystrophy; hematopoietic stem cell transplant; microglia; microglia replacement.

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Conflict of interest statement

Declaration of interests F.C.B. and M.L.B. are co-inventors on a patent filed by the board of trustees of the Leland Stanford Junior University (application 16/566,675), related to methods of microglia replacement. F.C.B. holds shares in NovoGlia Inc. M.L.B. holds shares in Alector Inc.

References

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Direct microglia replacement reveals pathologic and therapeutic contributions of brain macrophages to a monogenic neurological disease

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Direct microglia replacement reveals pathologic and therapeutic contributions of brain macrophages to a monogenic neurological disease

Authors

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Abstract

Conflict of interest statement

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