Studies On The Role Of Mitochondrial Dysfunction In Amyloid- β Mediated Neurodegeneration
EPrints@IICB
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Title |
Studies On The Role Of Mitochondrial Dysfunction In Amyloid- β Mediated Neurodegeneration |
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Creator |
DAS, HRISHITA
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Subject |
Cell Biology & Physiology
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Description |
Alzheimer’s disease (AD) is one of the major causes of dementia worldwide. AD is characterized clinically by progressive cognitive decline, and pathologically by the presence of extracellular senile plaques composed primarily of amyloid-β peptide (Aβ) and intracellular neurofibrillary tangles made up mainly of hyper phosphorylated tau.AD is a progressive disease and may takes more than twenty years to develop prominent clinical symptoms such as memory loss. It is important to unfold early molecular mechanisms that contribute to development of the disease.Aberrant accumulation of amyloid-β (Aβ) in brain is the major trigger for pathogenesis in AD. It is hence essential to understand how Aβ attains such toxic levels in the brain parenchyma. Aging is one of the major risk factors of AD. DNA damage plays an important role both in aging and AD. We studied the effect of both toxic and sub lethal dose of DNA damage on neurons. Treating differentiated SH-SY5Y cells with a toxic dose of Camptothecin(CPT) induced ROS, mitochondrial dysfunction and apoptosis. Increased expression of BID which is a BH3-only pro-apoptotic protein was observed under similar treatment. Down regulation of BID provided transient protection from toxic dose of CPT in neurons. Next we shifted our focus to study the stress response in neuron induced by sub lethal dose of DNA damaging agent CPT and Doxorubicin (Dox) without inducing cell death.We detected that a very subtle and tolerable amount of DNA damage, related to aging, increased intraneuronal Aβ1-42 production both in cultured neuron and in the cortex of rodent brain.Strikingly, we also observed elevated levels of mitochondrial fusion and of its major driver protein, MFN2. Hyperfusion of mitochondria may be seen as an adaptive stress response resulting from the induction of ER stress since we detected upregulated phosphorylation of both PERK, an ii | P a g e important arm of unfolded protein response and an ER-stress marker, and its substrate eukaryotic initiation factor 2 α (eIF2α), together constituting a signaling responsible for protective mitochondrial remodeling. This adaptive remodeling of mitochondria resulted in an increase in mitochondrial oxygen consumption rate and ATP production. At later time points this elongated structure of mitochondria shifts towards fission. Mitochondrial fission is observed in AD brain but the phenomenon that is leading to this shift is not well studied in the disease model. Reports suggest that eIF2α phosphorylation can increase BACE1 activity, the rate limiting enzyme in Aβ production. In our model, we show that inhibiting PERK, decreased Aβ1-42 level while direct BACE1 inhibition, reduced the mitochondrial fusion.Moreover, we found increased MFN2 expression in younger 5xFAD transgenic mice when Aβ plaques and neurodegeneration were absent. Down regulation of MFN2 decreased CPT induced increase in Aβ1-42in neuron. Thus, this study indicates that mild DNA damage leads to increased Aβ1-42 production via ER stress and may also direct mitochondrial elongation as an initial adaptive/protective response. We propose that age-related subtle genomic DNA damage may trigger enhanced intraneuronal Aβ1- 42 production in an apparently healthy neuron way before the appearance of clinical symptoms in AD. Mitochondrial genome can bear signatures for AD. Mutation in mitochondrial genome can compromise its function and also can be considered as biomarker for early diagnosis of the disease. In the next part of our work we have analysed point mutations in mtDNA from AD patient’s whole blood. We identified SNPs which are common and unique to all the samples. The nature and distribution of mutations were also studied. To our surprise the mutations were not due to oxidation, but it was due to replication error. The SNPs were dispersed in coding region not in D loop of mtDNA. Defective replication of mtDNA could be a result of ineffective replication carried by POLG. Further work in this direction will help to understand the reason of developing these mutations in AD patients’ whole blood. Altogether this work explores an unconventional role of mitochondria in AD. The connection of mitochondria in subtle DNA damage associated stress response and increase in Aβ1-42 production in neuronsis a novel finding and may lead to better understanding of the pathogenesis of AD. |
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Date |
2021
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Type |
Thesis
NonPeerReviewed |
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Format |
application/pdf
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Identifier |
http://www.eprints.iicb.res.in/2835/1/Thesis_for_Evaluation_Hrishita_Das.pdf
DAS, HRISHITA (2021) Studies On The Role Of Mitochondrial Dysfunction In Amyloid- β Mediated Neurodegeneration. PhD thesis, UNIVERSITY OF CALCUTTA. |
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Relation |
http://www.eprints.iicb.res.in/2835/
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