Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia
Microglia are key in the homeostatic well-being of the brain and microglial dysfunction has been implicated in neurodegenerative disorders such as Alzheimer’s disease (AD). Due to the many limitations to study microglia in situ or isolated for large scale drug discovery applications, there is a high...
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Format: | Article |
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Frontiers Media S.A.
2021-02-01
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Series: | Frontiers in Immunology |
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Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2020.617860/full |
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doaj-3bb1d84e6b0e42ef9025bb346883a564 |
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record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Marvin Reich Marvin Reich Iñaki Paris Iñaki Paris Martin Ebeling Nadine Dahm Christophe Schweitzer Dieter Reinhardt Roland Schmucki Megana Prasad Fabian Köchl Marcel Leist Sally A. Cowley Jitao David Zhang Christoph Patsch Simon Gutbier Markus Britschgi |
spellingShingle |
Marvin Reich Marvin Reich Iñaki Paris Iñaki Paris Martin Ebeling Nadine Dahm Christophe Schweitzer Dieter Reinhardt Roland Schmucki Megana Prasad Fabian Köchl Marcel Leist Sally A. Cowley Jitao David Zhang Christoph Patsch Simon Gutbier Markus Britschgi Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia Frontiers in Immunology iPSC (induced pluripotent stem cell) microglia cell culture protocols drug development TREM2 (triggering receptor expressed on myeloid cells) Alzheimer’s disease (AD) |
author_facet |
Marvin Reich Marvin Reich Iñaki Paris Iñaki Paris Martin Ebeling Nadine Dahm Christophe Schweitzer Dieter Reinhardt Roland Schmucki Megana Prasad Fabian Köchl Marcel Leist Sally A. Cowley Jitao David Zhang Christoph Patsch Simon Gutbier Markus Britschgi |
author_sort |
Marvin Reich |
title |
Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia |
title_short |
Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia |
title_full |
Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia |
title_fullStr |
Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia |
title_full_unstemmed |
Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived Microglia |
title_sort |
alzheimer’s risk gene trem2 determines functional properties of new type of human ipsc-derived microglia |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Immunology |
issn |
1664-3224 |
publishDate |
2021-02-01 |
description |
Microglia are key in the homeostatic well-being of the brain and microglial dysfunction has been implicated in neurodegenerative disorders such as Alzheimer’s disease (AD). Due to the many limitations to study microglia in situ or isolated for large scale drug discovery applications, there is a high need to develop robust and scalable human cellular models of microglia with reliable translatability to the disease. Here, we describe the generation of microglia-like cells from human induced pluripotent stem cells (iPSC) with distinct phenotypes for mechanistic studies in AD. We started out from an established differentiation protocol to generate primitive macrophage precursors mimicking the yolk sac ontogeny of microglia. Subsequently, we tested 36 differentiation conditions for the cells in monoculture where we exposed them to various combinations of media, morphogens, and extracellular matrices. The optimized protocol generated robustly ramified cells expressing key microglial markers. Bulk mRNA sequencing expression profiles revealed that compared to cells obtained in co-culture with neurons, microglia-like cells derived from a monoculture condition upregulate mRNA levels for Triggering Receptor Expressed On Myeloid Cells 2 (TREM2), which is reminiscent to the previously described disease-associated microglia. TREM2 is a risk gene for AD and an important regulator of microglia. The regulatory function of TREM2 in these cells was confirmed by comparing wild type with isogenic TREM2 knock-out iPSC microglia. The TREM2-deficient cells presented with stronger increase in free cytosolic calcium upon stimulation with ATP and ADP, as well as stronger migration towards complement C5a, compared to TREM2 expressing cells. The functional differences were associated with gene expression modulation of key regulators of microglia. In conclusion, we have established and validated a work stream to generate functional human iPSC-derived microglia-like cells by applying a directed and neuronal co-culture independent differentiation towards functional phenotypes in the context of AD. These cells can now be applied to study AD-related disease settings and to perform compound screening and testing for drug discovery. |
topic |
iPSC (induced pluripotent stem cell) microglia cell culture protocols drug development TREM2 (triggering receptor expressed on myeloid cells) Alzheimer’s disease (AD) |
url |
https://www.frontiersin.org/articles/10.3389/fimmu.2020.617860/full |
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doaj-3bb1d84e6b0e42ef9025bb346883a5642021-02-03T06:07:40ZengFrontiers Media S.A.Frontiers in Immunology1664-32242021-02-011110.3389/fimmu.2020.617860617860Alzheimer’s Risk Gene TREM2 Determines Functional Properties of New Type of Human iPSC-Derived MicrogliaMarvin Reich0Marvin Reich1Iñaki Paris2Iñaki Paris3Martin Ebeling4Nadine Dahm5Christophe Schweitzer6Dieter Reinhardt7Roland Schmucki8Megana Prasad9Fabian Köchl10Marcel Leist11Sally A. Cowley12Jitao David Zhang13Christoph Patsch14Simon Gutbier15Markus Britschgi16Roche Pharma Research and Early Development, Neuroscience and Rare Diseases Discovery and Translational Area, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandIn Vitro Toxicology and Biomedicine, Department inaugurated by the Doerenkamp-Zbinden Foundation, University of Konstanz, Konstanz, GermanyRoche Pharma Research and Early Development, Neuroscience and Rare Diseases Discovery and Translational Area, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandAchucarro Basque Center for Neuroscience, Science Park of the UPV/EHU, Leioa, SpainRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Therapeutic Modalities, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Neuroscience and Rare Diseases Discovery and Translational Area, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Neuroscience and Rare Diseases Discovery and Translational Area, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandIn Vitro Toxicology and Biomedicine, Department inaugurated by the Doerenkamp-Zbinden Foundation, University of Konstanz, Konstanz, GermanyJames Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, Oxford, United KingdomRoche Pharma Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Therapeutic Modalities, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Therapeutic Modalities, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandRoche Pharma Research and Early Development, Neuroscience and Rare Diseases Discovery and Translational Area, Roche Innovation Center Basel, F. Hoffmann-La Roche Ltd, Basel, SwitzerlandMicroglia are key in the homeostatic well-being of the brain and microglial dysfunction has been implicated in neurodegenerative disorders such as Alzheimer’s disease (AD). Due to the many limitations to study microglia in situ or isolated for large scale drug discovery applications, there is a high need to develop robust and scalable human cellular models of microglia with reliable translatability to the disease. Here, we describe the generation of microglia-like cells from human induced pluripotent stem cells (iPSC) with distinct phenotypes for mechanistic studies in AD. We started out from an established differentiation protocol to generate primitive macrophage precursors mimicking the yolk sac ontogeny of microglia. Subsequently, we tested 36 differentiation conditions for the cells in monoculture where we exposed them to various combinations of media, morphogens, and extracellular matrices. The optimized protocol generated robustly ramified cells expressing key microglial markers. Bulk mRNA sequencing expression profiles revealed that compared to cells obtained in co-culture with neurons, microglia-like cells derived from a monoculture condition upregulate mRNA levels for Triggering Receptor Expressed On Myeloid Cells 2 (TREM2), which is reminiscent to the previously described disease-associated microglia. TREM2 is a risk gene for AD and an important regulator of microglia. The regulatory function of TREM2 in these cells was confirmed by comparing wild type with isogenic TREM2 knock-out iPSC microglia. The TREM2-deficient cells presented with stronger increase in free cytosolic calcium upon stimulation with ATP and ADP, as well as stronger migration towards complement C5a, compared to TREM2 expressing cells. The functional differences were associated with gene expression modulation of key regulators of microglia. In conclusion, we have established and validated a work stream to generate functional human iPSC-derived microglia-like cells by applying a directed and neuronal co-culture independent differentiation towards functional phenotypes in the context of AD. These cells can now be applied to study AD-related disease settings and to perform compound screening and testing for drug discovery.https://www.frontiersin.org/articles/10.3389/fimmu.2020.617860/fulliPSC (induced pluripotent stem cell)microgliacell culture protocolsdrug developmentTREM2 (triggering receptor expressed on myeloid cells)Alzheimer’s disease (AD) |