Title

Investigating effects of primed astrocytes on microglia in Alzheimer’s disease pathology

DGIST Authors

Na Yeon Kim ; Jinsoo Seo ; Seong-Woon Yu

Advisor

서진수

Co-Advisor(s)

Seong-Woon Yu

Issued Date

2024

Awarded Date

2024-02-01

Citation

Na Yeon Kim. (2024). Investigating effects of primed astrocytes on microglia in Alzheimer’s disease pathology. doi: 10.22677/THESIS.200000724600

Type

Thesis

Description

Alzheimer's disease;Priming;Astrocytes;Microglia;Apolipoprotein E;Aβ

Table Of Contents

Ⅰ. Introduction 1
1.1 Neuroinflammatory environment in AD 1
1.2 Innate immune memory : priming effect 2
1.3 Apolipoprotein E4 (ApoE4) and AD 3
1.4 Human-induced pluripotent stem cell model (hiPSC) 3

Ⅱ. Materials and Methods 5
2.1 Cell culture 5
2.1.1 Human-induced pluripotent stem cell (hiPSC) 5
2.1.2 Neural progenitor cells (NPCs) differentiation 5
2.1.3 Astrocyte differentiation 6
2.1.4 Immortalized human microglia 6
2.2 Immune challenge and induction of priming 6
2.3 RNA extraction and cDNA synthesis 6
2.4 quantitative Real-time PCR (qRT-PCR) 7
2.5 Immunocytochemistry 7
2.6 Immunohistochemistry 8
2.7 Cytokine array analysis 8
2.8 Aβ42 uptake assay 9
2.9 Microglial Aβ42 clearance assay 9
2.10 Statistical analysis 10

Ⅲ. Results 11
3.1 Generation of isogenic hiPSC-derived astrocytes and characterization of their identify 11
3.2 Probing priming effect of hiPSC-derived astrocytes by the immune challenge 14
3.3 Aβ42-treated primed astrocytes upregulate the immune response 17
3.4 Aβ42-treated primed astrocytes induce enhancement of Aβ42 clearance of human microglia 20
3.5 APOE4 astrocytes have a priming effect 22
3.6 Primed APOE4 astrocytes enhance Aβ42 uptake but reduce immune response compared with primed APOE3 astrocytes 23
3.7 Aβ42-treated primed APOE4 astrocytes reduce Aβ42 clearance of human microglia 26
3.8 A 9-week-old wild-type (C57BL/6J) male mouse has a priming effect, as in vitro 29
3.9 Confirming the priming effect and Aβ42 clearance of the 7-month-old humanized APOEKI 5XFAD male mouse 33
3.10 Confirming the priming effect and Aβ42 clearance of the 6-month-old humanized APOE KI male mouse 41

Ⅵ. Discussion 44

Ⅴ. References 47

Abstract in Korean 50

List of tables and figures

Table 1. Information of cell lines used in the experiment

Figure 1. Generation of the isogenic hiPSC-derived astrocytes and confirmation of their identify

Figure 2. hiPSC-derived astrocytes have a priming effect

Figure 3. Aβ42-treated primed astrocytes upregulate the immune response

Figure 4. The effect of primed astrocytes on microglia in AD context

Figure 5. APOE4 astrocytes have a priming effect and Aβ42-treated primed APOE4 astrocytes reduce immune response compared with Aβ42-treated primed APOE3 astrocytes

Figure 6. The effect of primed APOE4 astrocytes on microglia in AD context

Figure 7. A wild-type male mouse astrocyte has a priming effect

Figure 8. Confirming the priming effect and Aβ42 clearance of the humanized APOE KI 5XFAD male mouse

Figure 9. Confirming the priming effect and Aβ42 clearance of the humanized APOE KI male mouse

Figure 10. Graphic Summary

URI

http://hdl.handle.net/20.500.11750/48051
http://dgist.dcollection.net/common/orgView/200000724600

DOI

10.22677/THESIS.200000724600

Degree

Master

Department

Department of Brain Sciences

Publisher

DGIST

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