Title

Investigating the role of PTP1B in neurodegenerative mechanisms induced by ALS/FTD-associated RNA-binding proteins

DGIST Authors

Seyeon Kim ; Sung Bae Lee ; Hyung-Jun Kim

Advisor

이성배

Co-Advisor(s)

Hyung-Jun Kim

Issued Date

2025

Awarded Date

2025-02-01

Citation

Seyeon Kim. (2025). Investigating the role of PTP1B in neurodegenerative mechanisms induced by ALS/FTD-associated RNA-binding proteins. doi: 10.22677/THESIS.200000837594

Type

Thesis

Description

Neurodegenerative diseases, RBP, Neuroinflammation, Protein quality contorl, PTP1B

Table Of Contents

Chapter Ⅰ. Introduction 1
1.1 Research background and purpose 1
1.2 Neurodegenerative disease (ALS and FTD) 1
1.3 RNA binding protein associated with ALS and FTD 4
1.4 Neuroinflammation 8
1.5 Cellular processes of protein quality control system 8

Chapter Ⅱ. Materials & Methods 11
2.1 Reagents and Antibodies 11
2.2 Primary cell cultures 11
2.3 NSC-34 cell culture 12
2.4 Cell line 12
2.5 Transfection 13
2.6 Cytotoxicity test 13
2.7 CMFDA staining analysis 14
2.8 Astrocyte-conditioned media (ACM) 14
2.9 Cortical neuron-astrocytes co-culture assay 15
2.10 ACM-treated neuron culture 15
2.11 Quantitative RT-PCR 15
2.12 Immunoblot analysis 16
2.13 Nuclear and cytoplasmic fraction extraction 16
2.14 Preparation of soluble and insoluble cell extracts 16
2.15 ELISA 17
2.16 Immunocytochemistry analysis 17
2.17 Mitochondrial activity assay 17
2.18 Quantification of dendritic spines 18
2.19 Human primary fibroblasts 18
2.20 Flow Cytometry Analysis 19
2.21 CYTO-ID® Autophagy Analysis 19
2.22 Drosophila Immunohistochemical analysis 19
2.23 Fly stains 20
2.24 Lifespan and climbing assays 20
2.25 Statistics 20

Chapter Ⅲ. Results 21
Part 1. Non-cell autonomous effects of astrocytes in contributing to neuronal cell death 21
Part 1-1. The role of PTP1B inhibition in alleviating neuroinflammation associated with TDP-43 22
3.1 Investigating the role of PTP1B in CNS 22
3.2 PTP1B is an essential modulator of TDP-43-induced inflammation in astrocytes 24
3.3 PTP1B regulates TDP-43-induced inflammation via the NF-κB pathway 32
3.4 PTP1B inhibition and absorption of proinflammatory cytokines mitigate the neuronal toxicity caused by astrocytic TDP-43 overexpression 37
3.5 PTP1B inhibition suppresses astrocytic TDP-43-induced mitochondrial dysfunction and spine retraction in neurons 45
3.6 PTP1B inhibition ameliorates astrocytic TDP-43-induced neuronal toxicity and mitochondrial dysfunction in motor neuron-like cells 50
3.7 Inflammation and neuronal toxicity induced by glial TDP-43 are mitigated by PTP1B downregulation in Drosophila 55
Part 1-2. The effects of DHE in mitigating neuroinflammation related to mutant FUS in astrocytes 65
3.8 Investigating the role of natural compound-derived PTP1B inhibitors in CNS 65
3.9 DHE alleviates aggregation of FUS P525L and activation of inflammatory response in mutant FUS P525L overexpressed astrocytes 66
3.10 DHE treatment reduces astrocytic FUS P525L-induced neuronal toxicity and mitochondria dysfunction 73
3.11 DHE treatment rescues glial FUS induced toxicity in Drosophila 76
3.12 DHE treatment moderates inflammatory response in FUSALS patient fibroblast 83
Part 2. Cell-autonomous effects of TDP-43 on UPS impairment 88
3.13 The mitigating effects of PTP1B inhibition on UPS impairment dysregulation in neuronal cells 89
3.14 Induced ALP by TDP-43 overexpression is restored though inhibition of PTP1B in N2a 96
3.15 Inhibition of PTP1B ameliorates TDP-43-induced neurotoxicity in vitro and in vivo 101
3.16 PTP1B overexpression exacerbates TDP-43-induced neurotoxicity 106

Chapter Ⅳ. Discussion 113
References 120
Abstracts in Koreans 133

URI

http://hdl.handle.net/20.500.11750/57956
http://dgist.dcollection.net/common/orgView/200000837594

DOI

10.22677/THESIS.200000837594

Degree

Doctor

Department

Department of Brain Sciences

Publisher

DGIST

Show Full Item Record