INDÜKLENMIŞ PLURIPOTENT KÖK HÜCRE KAYNAKLI MIKROGLIA HÜCRELERININ ALZHEIMER HASTALIǦINDA NLRP3 INFLAMAZOM AKTIVASYONU ARAŞTIRMALARINDA KULLANIMI

Erdoǧan Öner, Asli; Dansokho, Cira; Koistinaho, Jari; Lehtonen, Sarka; Solakoǧlu, Seyhun; HENEKA, Michael

doi:10.26650/IUITFD.1451011

Article (Scientific journals)

INDÜKLENMIŞ PLURIPOTENT KÖK HÜCRE KAYNAKLI MIKROGLIA HÜCRELERININ ALZHEIMER HASTALIǦINDA NLRP3 INFLAMAZOM AKTIVASYONU ARAŞTIRMALARINDA KULLANIMI

Erdoǧan Öner, Asli; Dansokho, Cira; Koistinaho, Jari et al.

2024 • In Istanbul Tip Fakultesi Dergisi, 87 (4), p. 299 - 310

Peer reviewed

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https://hdl.handle.net/10993/67428

DOI
10.26650/IUITFD.1451011

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Keywords :

amyloid beta; induced pluripotent stem cells; Microglia; neuroinflammation; NLRP3 inflammasome; Medicine (all)

Abstract :

[en] Objective: Alzheimer's disease (AD) is an irreversible and progressive neurodegenerative disease. Besides amyloid beta (Aβ) and tau accumulations, inflammation also contributes to AD pathogenesis. NLR family pyrin domain containing 3 (NLRP3) inflammasome activation in microglia is thought to be associated with AD. Since animal models of AD do not accurately reflect human pathology, in our study, human induced pluripotent stem cells (iPSCs) were differentiated into microglia, and their potential to be used in NLRP3 pathway-related mechanisms in AD was investigated. Material and Method: iPSC cell lines of AD, isogenic, or control genotypes were differentiated into microglia and cells from different stages of the differentiation were characterized by flow cytometry, real-time quantitative polymerase chain reaction (RTqPCR), immunocytochemistry, and Western blot. The expression of proteins associated with the NLRP3 pathway was investigated by Western blot. For functional analysis, cytokine release was assessed by Enzyme-linked immunosorbent assay (ELISA) upon NLRP3 inflammasome activators (lipopolysaccharide (LPS), Aβ) and inhibitor (cytokine release inhibitory drug 3, CRID3) treatments. The phagocytosis of pHrodo particles and Aβ were evaluated by flow cytometry, fluorescence microscopy, and live cell imaging system. Result: In our study, we could differentiate microglia from iPSCs derived from different genotypes. These microglia cells expressed various microglia and NLRP3 inflammasome-related markers and were able to phagocytose pHrodo particles and Aβ. The stimulation of the microglia cells with LPS and Aβ caused IL-1beta and IL-18 release, while CRID3 reversed this effect. Conclusion: Our results show that iPSC-derived microglia generated in this study recapitulate microglia functional characteristics and can therefore be used to study NLRP3 pathway-associated disease mechanisms and treatment options.

Disciplines :

Neurology

Author, co-author :

Erdoǧan Öner, Asli ; Istanbul University, Istanbul Faculty of Medicine, Department of Histology and Embryology, Istanbul, Turkey ; Izmir Katip Çelebi University, Faculty of Medicine, Department of Histology and Embryology, Izmir, Turkey

Dansokho, Cira ; Evotec SE, Hamburg, Germany

Koistinaho, Jari ; Neuroscience Center, HiLIFE, University of Helsinki, Helsinki, Finland

Lehtonen, Sarka ; A.I. Virtanen Institute for Molecular Sciences, University of Eastern Finland, Kuopio, Finland

Solakoǧlu, Seyhun ; Istanbul University, Istanbul Faculty of Medicine, Department of Histology and Embryology, Istanbul, Turkey

HENEKA, Michael ; University of Luxembourg > Luxembourg Centre for Systems Biomedicine (LCSB)

External co-authors :

yes

Language :

Turkish

Title :

INDÜKLENMIŞ PLURIPOTENT KÖK HÜCRE KAYNAKLI MIKROGLIA HÜCRELERININ ALZHEIMER HASTALIǦINDA NLRP3 INFLAMAZOM AKTIVASYONU ARAŞTIRMALARINDA KULLANIMI

Publication date :

2024

Journal title :

Istanbul Tip Fakultesi Dergisi

ISSN :

1305-6433

eISSN :

1305-6441

Publisher :

Istanbul University Press

Volume :

Issue :

Pages :

299 - 310

Peer reviewed :

Peer reviewed

Funding text :

Financial Disclosure: This study was funded by T\u00DCB\u0130TAK (2214/A International Research Fellowship Programme for PhD Students, Project No: 1059B141900285), Y\u00D6K-YUDAB (Research Scholarship for Doctoral Studies Abroad for Research Assistants).

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Bibliography

Li T, Lu L, Pember E, Li X, Zhang B, Zhu Z. New Insights into neuroinflammation involved in pathogenic mechanism of Alzheimer's disease and its potential for therapeutic intervention. Cells 2022;11:1925. [CrossRef]
2022 Alzheimer's disease facts and figures. Alzheimer's and Dementia 2022;18(4):700-89. [CrossRef]
McManus RM. The Role of Immunity in Alzheimer's Disease. Adv Biol 2022;6(5):2101166. [CrossRef]
Crews L, Masliah E. Molecular mechanisms of neurodegeneration in Alzheimer's disease. Hum Mol Genet 2010;19(R1):R12-20. [CrossRef]
Heneka MT, Carson MJ, Khoury J El, Landreth GE, Brosseron F, Feinstein DL, et al. Neuroinflammation in Alzheimer's disease. Lancet Neurol 2015;14(4):388-405. [CrossRef]
Heppner FL, Ransohoff RM, Becher B. Immune attack: The role of inflammation in Alzheimer disease. Nat Rev Neurosci 2015;16(6):358-72. [CrossRef]
Li JW, Zong Y, Cao XP, Tan L, Tan L. Microglial priming in Alzheimer's disease. Ann Transl Med 2018;6(10):176. [CrossRef]
Kettenmann H, Hanisch UK, Noda M, Verkhratsky A. Physiology of microglia. Physiol Rev 2011;91(2):461-553. [CrossRef]
Fan Y, Xie L, Chung CY. Signaling pathways controlling microglia chemotaxis. Mol Cells 2017;40(3):163-8. [CrossRef]
Heneka MT, Kummer MP, Stutz A, Delekate A, Schwartz S, Vieira-Saecker A, et al. NLRP3 is activated in Alzheimer's disease and contributes to pathology in APP/PS1 mice. Nature 2013;493(7434):674-8. [CrossRef]
Rui W, Xiao H, Fan Y, Ma Z, Xiao M, Li S, et al. Systemic inflammasome activation and pyroptosis associate with the progression of amnestic mild cognitive impairment and Alzheimer's disease. J Neuroinflammation 2021;18(1):280. [CrossRef]
Dansokho C, Heneka MT. Neuroinflammatory responses in Alzheimer's disease. J Neural Transm 2018;125(5):771-9. [CrossRef]
Swanson K, Deng M, Ting JPY. The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol 2019;19:477-89. [CrossRef]
Liang T, Zhang Y, Wu S, Chen Q, Wang L. The Role of NLRP3 Inflammasome in Alzheimer's Disease and Potential Therapeutic Targets. Front Pharmacol 2013;13:845185. [CrossRef]
Daniels MJD, Rivers-Auty J, Schilling T, Spencer NG, Watremez W, Fasolino V, et al. Fenamate NSAIDs inhibit the NLRP3 inflammasome and protect against Alzheimer's disease in rodent models. Nat Commun 2016;7:12504. [CrossRef]
Yin J, Zhao F, Chojnacki JE, Fulp J, Klein WL, Zhang S, et al. NLRP3 inflammasome inhibitor ameliorates amyloid pathology in a mouse model of Alzheimer's disease. Mol Neurobiol 2018;55(3):1977-87. [CrossRef]
Zahid A, Li B, Kombe AJK, Jin T, Tao J. Pharmacological inhibitors of the NLRP3 inflammasome. Front Immunol 2019;10:2538. [CrossRef]
Coll RC, Hill JR, Day CJ, Zamoshnikova A, Boucher D, Massey NL, et al. MCC950 directly targets the NLRP3 ATP-hydrolysis motif for inflammasome inhibition. Nat Chem Biol 2019;15(6):556-9. [CrossRef]
Barczuk J, Siwecka N, Lusa W, Rozpȩdek-Kamińska W, Kucharska E, Majsterek I. Targeting NLRP3-Mediated Neuroinflammation in Alzheimer's Disease Treatment. Int Journal of Mol Sci 2022;23(16):8979. [CrossRef]
Galatro TF, Holtman IR, Lerario AM, Vainchtein ID, Brouwer N, Sola PR, et al. Transcriptomic analysis of purified human cortical microglia reveals age-associated changes. Nat Neurosci 2017;20(8):1162-71. [CrossRef]
Washer SJ, Perez-Alcantara M, Chen Y, Steer J, James WS, Trynka G, et al. Single-cell transcriptomics defines an improved, validated monoculture protocol for differentiation of human iPSC to microglia. Sci Rep 2022;12(1):19454. [CrossRef]
Hasselmann J, Blurton-Jones M. Human iPSC-derived microglia: A growing toolset to study the brain's innate immune cells. Glia 2020;68(4):721-39. [CrossRef]
McQuade A, Coburn M, Tu CH, Hasselmann J, Davtyan H, Blurton-Jones M. Development and validation of a simplified method to generate human microglia from pluripotent stem cells. Mol Neurodegener 2018;13:67. [CrossRef]
Leng F, Edison P. Neuroinflammation and microglial activation in Alzheimer disease: where do we go from here? Nat Rev Neurol 2021;17:157-72. [CrossRef]
Pocock JM, Piers TM. Modelling microglial function with induced pluripotent stem cells: An update. Nat Rev Neurosci 2018;19(8):445-52. [CrossRef]
Friedman BA, Srinivasan K, Ayalon G, Meilandt WJ, Lin H, Huntley MA, et al. Diverse brain myeloid expression profiles reveal distinct microglial activation states and aspects of Alzheimer's disease not evident in mouse models. Cell Rep 2018;22(3):832-47. [CrossRef]
Kierdorf K, Erny D, Goldmann T, Sander V, Schulz C, Perdiguero EG, et al. Microglia emerge from erythromyeloid precursors via Pu.1-and Irf8-dependent pathways. Nat Neurosci 2013;16(3):273-80. [CrossRef]
Ginhoux F, Greter M, Leboeuf M, Nandi S, See P, Gokhan S, et al. Fate mapping analysis reveals that adult microglia derive from primitive macrophages. Science 2010;330(6005):841-5. [CrossRef]
Haenseler W, Sansom SN, Buchrieser J, Newey SE, Moore CS, Nicholls FJ, et al. A Highly Efficient Human Pluripotent Stem Cell Microglia Model Displays a Neuronal-Co-culture- Specific Expression Profile and Inflammatory Response. Stem Cell Rep 2017;8(6):1727-42. [CrossRef]
Pandya H, Shen MJ, Ichikawa DM, Sedlock AB, Choi Y, Johnson KR, et al. Differentiation of human and murine induced pluripotent stem cells to microglia-like cells. Nat Neurosci 2017;20(5):753-9. [CrossRef]
Takata K, Kozaki T, Lee CZW, Thion MS, Otsuka M, Lim S, et al. Induced-Pluripotent-Stem-Cell-Derived Primitive Macrophages Provide a Platform for Modeling Tissue- Resident Macrophage Differentiation and Function. Immunity 2017;47(1):183-98. [CrossRef]
Abud EM, Ramirez RN, Martinez ES, Healy LM, Nguyen CHH, Newman SA, et al. iPSC-derived human microglia-like cells to study neurological diseases. Neuron 2017;94(2):278- 93. [CrossRef]
Konttinen H, Cabral-da-Silva M e. C, Ohtonen S, Wojciechowski S, Shakirzyanova A, Caligola S, et al. PSEN1ΔE9, APPswe, and APOE4 Confer Disparate Phenotypes in Human iPSC-Derived Microglia. Stem Cell Rep 2019;13(4):669-83. [CrossRef]
Douvaras P, Sun B, Wang M, Kruglikov I, Lallos G, Zimmer M, et al. Directed Differentiation of Human Pluripotent Stem Cells to Microglia. Stem Cell Rep 2017;8(6):1516-24. [CrossRef]
Foudah D, Monfrini M, Donzelli E, Niada S, Brini AT, Orciani M, et al. Expression of neural markers by undifferentiated mesenchymal-like stem cells from different sources. J Immunol Res 2014;2014:987678. [CrossRef]
Bianchi F, Malboubi M, Li Y, George JH, Jerusalem A, Szele F, et al. Rapid and efficient differentiation of functional motor neurons from human iPSC for neural injury modelling. Stem Cell Res 2018;32:126-34. [CrossRef]
Böttcher C, Schlickeiser S, Sneeboer MAM, Kunkel D, Knop A, Paza E, et al. Human microglia regional heterogeneity and phenotypes determined by multiplexed single-cell mass cytometry. Nat Neurosci 2019;22(1):78-90. [CrossRef]
Melief J, Sneeboer MAM, Litjens M, Ormel PR, Palmen SJMC, Huitinga I, et al. Characterizing primary human microglia: A comparative study with myeloid subsets and culture models. Glia 2016;64(11):1857-68. [CrossRef]
Garcia-Reitboeck P, Phillips A, Piers TM, Villegas-Llerena C, Butler M, Mallach A, et al. Human Induced Pluripotent Stem Cell-Derived Microglia-Like Cells Harboring TREM2 Missense Mutations Show Specific Deficits in Phagocytosis. Cell Rep 2018;24(9):2300-11. [CrossRef]
Satoh J, Kino Y, Asahina N, Takitani M, Miyoshi J, Ishida T, et al. TMEM119 marks a subset of microglia in the human brain. Neuropathology 2016;36(1):39-49. [CrossRef]
Bohnert S, Seiffert A, Trella S, Bohnert M, Distel L, Ondruschka B, et al. TMEM119 as a specific marker of microglia reaction in traumatic brain injury in postmortem examination. Int J Legal Med 2020;134(6):2167-76. [CrossRef]
Murai N, Mitalipova M, Jaenisch, R. Functional analysis of CX3CR1 in human induced pluripotent stem (iPS) cell-derived microglia-like cells. Eur J Neurosci 2020;52(7):3667- 78. [CrossRef]
Morillas AG, Besson VC, Lerouet D. Microglia and neuroinflammation: What place for p2ry12? Int J Mol Sci 2021;22(4):1-16. [CrossRef]
Lučiunaite A, McManus RM, Jankunec M, Rácz I, Dansokho C, Dalgediene I, et al. Soluble Aβ oligomers and protofibrils induce NLRP3 inflammasome activation in microglia. J Neurochem 2020;155(6):650-61. [CrossRef]
Dempsey C, Rubio Araiz A, Bryson KJ, Finucane O, Larkin C, Mills EL, et al. Inhibiting the NLRP3 inflammasome with MCC950 promotes non-phlogistic clearance of amyloid-β and cognitive function in APP/PS1 mice. Brain Behav Immun 2017;61:306-16. [CrossRef]
Mouton-Liger F, Rosazza T, Sepulveda-Diaz J, Ieang A, Hassoun SM, Claire E, et al. Parkin deficiency modulates NLRP3 inflammasome activation by attenuating an A20- dependent negative feedback loop. Glia 2018;66(8):1736- 51. [CrossRef]
Clénet ML, Keaney J, Gillet G, Valadas JS, Langlois J, Cardenas A, et al. Divergent functional outcomes of NLRP3 blockade downstream of multi-inflammasome activation: therapeutic implications for ALS. Front Immunol 2023;14:1190219. [CrossRef]
Baik SH, Kang S, Son SM, Mook-Jung I. Microglia contributes to plaque growth by cell death due to uptake of amyloid β in the brain of Alzheimer's disease mouse model. Glia 2016;64(12):2274-90. [CrossRef]
Huang Y, Happonen KE, Burrola PG, O'Connor C, Hah N, Huang L, et al. Microglia use TAM receptors to detect and engulf amyloid β plaques. Nat Immunol 2021;22(5):586-94. [CrossRef]
Xu M, Zhang L, Liu G, Jiang N, Zhou W, Zhang Y. Pathological Changes in Alzheimer's Disease Analyzed Using Induced Pluripotent Stem Cell-Derived Human Microglia-Like Cells. J Alzheimer's Dis 2019;67(1):357-68. [CrossRef]