Amyloid-β precursor protein processing and oxidative stress are altered in human iPSC-derived neuron and astrocyte co-cultures carrying presenillin-1 gene mutations following spontaneous differentiation
Elsworthy, Richard J ; King, Marianne C ; Grainger, Alastair ; Fisher, Emily ; Crowe, James A LU
; Alqattan, Sarah
; Ludlam, Adele
; Hill, Dr Eric J
and Aldred, Sarah
(2021)
In Molecular and Cellular Neuroscience
114.
- Abstract
INTRODUCTION: Presenilin-1 (PSEN1) gene mutations are the most common cause of familial Alzheimer's disease (fAD) and are known to interfere with activity of the membrane imbedded γ-secretase complex. PSEN1 mutations have been shown to shift Amyloid-β precursor protein (AβPP) processing toward amyloid-β (Aβ) 1-42 production. However, less is known about whether PSEN1 mutations may alter the activity of enzymes such as ADAM10, involved with non-amyloidogenic AβPP processing, and markers of oxidative stress.
MATERIALS AND METHODS: Control and PSEN1 mutation (L286V and R278I) Human Neural Stem Cells were spontaneously differentiated into neuron and astrocyte co-cultures. Cell lysates and culture media were collected and stored at -80... (More)
INTRODUCTION: Presenilin-1 (PSEN1) gene mutations are the most common cause of familial Alzheimer's disease (fAD) and are known to interfere with activity of the membrane imbedded γ-secretase complex. PSEN1 mutations have been shown to shift Amyloid-β precursor protein (AβPP) processing toward amyloid-β (Aβ) 1-42 production. However, less is known about whether PSEN1 mutations may alter the activity of enzymes such as ADAM10, involved with non-amyloidogenic AβPP processing, and markers of oxidative stress.
MATERIALS AND METHODS: Control and PSEN1 mutation (L286V and R278I) Human Neural Stem Cells were spontaneously differentiated into neuron and astrocyte co-cultures. Cell lysates and culture media were collected and stored at -80 °C until further analysis. ADAM10 protein expression, the ratio of AβPP forms and Aβ1-42/40 were assessed. In addition, cellular redox status was quantified.
RESULTS: The ratio of AβPP isoforms (130:110kDa) was significantly reduced in neuron and astrocyte co-cultures carrying PSEN1 gene mutations compared to control, and mature ADAM10 expression was lower in these cells. sAβPP-α was also significantly reduced in L286V mutation, but not in the R278I mutation cells. Both Aβ1-40 and Aβ1-42 were increased in conditioned cell media from L286V cells, however, this was not matched in R278I cells. The Aβ1-42:40 ratio was significantly elevated in R278I cells. Markers of protein carbonylation and lipid peroxidation were altered in both l286V and R278I mutations. Antioxidant status was significantly lower in R278I cells compared to control cells.
CONCLUSIONS: This data provides evidence that the PSEN1 mutations L286V and R278I significantly alter protein expression associated with AβPP processing and cellular redox status. In addition, this study highlights the potential for iPSC-derived neuron and astrocyte co-cultures to be used as an early human model of fAD.
(Less)
- author
- Elsworthy, Richard J
; King, Marianne C
; Grainger, Alastair
; Fisher, Emily
; Crowe, James A
LU
; Alqattan, Sarah
; Ludlam, Adele
; Hill, Dr Eric J
and Aldred, Sarah
- publishing date
- 2021-07
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Molecular and Cellular Neuroscience
- volume
- 114
- article number
- 103631
- publisher
- Elsevier
- external identifiers
-
- scopus:85107784969
- pmid:34022327
- ISSN
- 1044-7431
- DOI
- 10.1016/j.mcn.2021.103631
- language
- English
- LU publication?
- no
- additional info
- Copyright © 2021 Elsevier Inc. All rights reserved.
- id
- 19b9d013-90c8-4d05-a930-3e2559b41dc1
- date added to LUP
- 2021-12-14 13:48:09
- date last changed
- 2024-10-20 12:12:29
@article{19b9d013-90c8-4d05-a930-3e2559b41dc1,
abstract = {{<p>INTRODUCTION: Presenilin-1 (PSEN1) gene mutations are the most common cause of familial Alzheimer's disease (fAD) and are known to interfere with activity of the membrane imbedded γ-secretase complex. PSEN1 mutations have been shown to shift Amyloid-β precursor protein (AβPP) processing toward amyloid-β (Aβ) 1-42 production. However, less is known about whether PSEN1 mutations may alter the activity of enzymes such as ADAM10, involved with non-amyloidogenic AβPP processing, and markers of oxidative stress.</p><p>MATERIALS AND METHODS: Control and PSEN1 mutation (L286V and R278I) Human Neural Stem Cells were spontaneously differentiated into neuron and astrocyte co-cultures. Cell lysates and culture media were collected and stored at -80 °C until further analysis. ADAM10 protein expression, the ratio of AβPP forms and Aβ1-42/40 were assessed. In addition, cellular redox status was quantified.</p><p>RESULTS: The ratio of AβPP isoforms (130:110kDa) was significantly reduced in neuron and astrocyte co-cultures carrying PSEN1 gene mutations compared to control, and mature ADAM10 expression was lower in these cells. sAβPP-α was also significantly reduced in L286V mutation, but not in the R278I mutation cells. Both Aβ1-40 and Aβ1-42 were increased in conditioned cell media from L286V cells, however, this was not matched in R278I cells. The Aβ1-42:40 ratio was significantly elevated in R278I cells. Markers of protein carbonylation and lipid peroxidation were altered in both l286V and R278I mutations. Antioxidant status was significantly lower in R278I cells compared to control cells.</p><p>CONCLUSIONS: This data provides evidence that the PSEN1 mutations L286V and R278I significantly alter protein expression associated with AβPP processing and cellular redox status. In addition, this study highlights the potential for iPSC-derived neuron and astrocyte co-cultures to be used as an early human model of fAD.</p>}},
author = {{Elsworthy, Richard J and King, Marianne C and Grainger, Alastair and Fisher, Emily and Crowe, James A and Alqattan, Sarah and Ludlam, Adele and Hill, Dr Eric J and Aldred, Sarah}},
issn = {{1044-7431}},
language = {{eng}},
publisher = {{Elsevier}},
series = {{Molecular and Cellular Neuroscience}},
title = {{Amyloid-β precursor protein processing and oxidative stress are altered in human iPSC-derived neuron and astrocyte co-cultures carrying presenillin-1 gene mutations following spontaneous differentiation}},
url = {{http://dx.doi.org/10.1016/j.mcn.2021.103631}},
doi = {{10.1016/j.mcn.2021.103631}},
volume = {{114}},
year = {{2021}},
}