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
Autor: | Adele Ludlam, Eric J. Hill, Marianne C. King, Sarah Alqattan, Emily Fisher, James A Crowe, Richard J Elsworthy, Alastair Grainger, Sarah Aldred |
---|---|
Rok vydání: | 2021 |
Předmět: |
0301 basic medicine
Gene isoform ADAM10 Induced Pluripotent Stem Cells Gene mutation Biology medicine.disease_cause Presenilin 03 medical and health sciences Cellular and Molecular Neuroscience Amyloid beta-Protein Precursor 0302 clinical medicine medicine PSEN1 Presenilin-1 Humans Molecular Biology Neurons Mutation Amyloid beta-Peptides Cell Differentiation Cell Biology Neural stem cell Coculture Techniques Cell biology Oxidative Stress 030104 developmental biology medicine.anatomical_structure Astrocytes 030217 neurology & neurosurgery Astrocyte |
Zdroj: | Molecular and cellular neurosciences. 114 |
ISSN: | 1095-9327 |
Popis: | 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. |
Databáze: | OpenAIRE |
Externí odkaz: |