Methods related to studying tau fragmentation

Ferreira, Adriana; Afreen, Sana

doi:10.1016/bs.mcb.2017.06.004

Cited by 6 publications

(3 citation statements)

References 31 publications

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“…Tau proteins, mutant and wild-type, in human brain are heterogeneous in molecular forms with respect to different repeat domains and proteolytic fragments (1,(127)(128)(129)(130)(131)(132). The finding in this study that the P301L and V337M Tau mutations result in dysregulation of exosome protein cargo leads the question of what are the effects of other Tau isoforms on exosome cargoes and pathogenicity?…”

Section: Molecular and Cellular Proteomics 196 1027mentioning

confidence: 78%

Dysregulation of Exosome Cargo by Mutant Tau Expressed in Human-induced Pluripotent Stem Cell (iPSC) Neurons Revealed by Proteomics Analyses

Podvin

Jones

Liu

et al. 2020

Molecular & Cellular Proteomics

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Accumulation and propagation of hyperphosphorylated Tau (p-Tau) is a common neuropathological hallmark associated with neurodegeneration of Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), and related tauopathies. Extracellular vesicles, specifically exosomes, have recently been demonstrated to participate in mediating Tau propagation in brain. Exosomes produced by human induced pluripotent stem cell (iPSC)-derived neurons expressing mutant Tau (mTau), containing the P301L and V337M Tau mutations of FTDP-17, possess the ability to propagate p-Tau pathology after injection into mouse brain. To gain an understanding of the mTau exosome cargo involved in Tau pathogenesis, these pathogenic exosomes were analyzed by proteomics and bioinformatics. The data showed that mTau expression dysregulates the exosome proteome to result in 1) proteins uniquely present only in mTau, and not control exosomes, 2) the absence of proteins in mTau exosomes, uniquely present in control exosomes, and 3) shared proteins which were significantly upregulated or downregulated in mTau compared with control exosomes. Notably, mTau exosomes (not control exosomes) contain ANP32A (also known as I1PP2A), an endogenous inhibitor of the PP2A phosphatase which regulates the phosphorylation state of p-Tau. Several of the mTau exosome-specific proteins have been shown to participate in AD mechanisms involving lysosomes, inflammation, secretases, and related processes. Furthermore, the mTau exosomes lacked a substantial portion of proteins present in control exosomes involved in pathways of localization, vesicle transport, and protein binding functions. The shared proteins present in both mTau and control exosomes represented exosome functions of vesicle-mediated transport, exocytosis, and secretion processes. These data illustrate mTau as a dynamic regulator of the biogenesis of exosomes to result in acquisition, deletion, and up- or downregulation of protein cargo to result in pathogenic mTau exosomes capable of in vivo propagation of p-Tau neuropathology in mouse brain.

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Section: Molecular and Cellular Proteomics 196 1027mentioning

confidence: 78%

Dysregulation of Exosome Cargo by Mutant Tau Expressed in Human-induced Pluripotent Stem Cell (iPSC) Neurons Revealed by Proteomics Analyses

Podvin

Jones

Liu

et al. 2020

Molecular & Cellular Proteomics

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“…It is well known that in tumor cells undergoing apoptosis, the DNA is cleaved at the site of attachment resulting in DNA fragments with free 3 0 -OH end [111]. This facilitates identification of apoptotic cells by transfer of fluorescent-labeled dUTP to the free 3 0 -OH ends using the enzyme terminal deoxynucleotidyl transferase (TdT) [112]. Interestingly, our TUNEL results had depicted that the quadruple-combinatorial treatment (TCNP) had significantly induced apoptosis as evident from the highest percentage of TUNEL-positive cells with brown and shrunken nuclei (Figure 4b, red arrowheads).…”

Section: Discussionmentioning

confidence: 99%

Repurposing synthetic and natural derivatives induces apoptosis in an orthotopic glioma‐induced xenograft model by modulating WNT/β‐catenin signaling

Precilla

Kuduvalli

Biswas

et al. 2023

Fundamemntal Clinical Pharma

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BackgroundGlioblastomas arise from multistep tumorigenesis of the glial cells. Despite the current state‐of‐art treatment, tumor recurrence is inevitable. Among the innovations blooming up against glioblastoma, drug repurposing could provide profound premises for treatment enhancement. While considering this strategy, the efficacy of the repurposed drugs as monotherapies were not up to par; hence, the focus has now shifted to investigate the multidrug combinations.AimTo investigate the efficacy of a quadruple‐combinatorial treatment comprising temozolomide along with chloroquine, naringenin, and phloroglucinol in an orthotopic glioma‐induced xenograft model.MethodsAntiproliferative effect of the drugs was assessed by immunostaining. The expression profiles of WNT/β‐catenin and apoptotic markers were evaluated by qRT‐PCR, immunoblotting, and ELISA. Patterns of mitochondrial depolarization was determined by flow cytometry. TUNEL assay was performed to affirm apoptosis induction. In vivo drug detection study was carried out by ESI‐Q‐TOF MS analysis.ResultsThe quadruple‐drug treatment had significantly hampered glioma proliferation and had induced apoptosis by modulating the WNT/β‐catenin signaling. Interestingly, the induction of apoptosis was associated with mitochondrial depolarization. The quadruple‐drug cocktail had breached the blood–brain barrier and was detected in the brain tissue and plasma samples.ConclusionThe quadruple‐drug combination served as a promising adjuvant therapy to combat glioblastoma lethality in vivo and can be probed for translation from bench to bedside.

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“…It is well-known that in tumor cells undergoing apoptosis, the DNA is cleaved at the site of attachment resulting in DNA fragments with free 3’-OH end (Elmore 2007). This facilitates identification of apoptotic cells by transfer of fluorescent-labelled dUTP to the free 3’-OH ends using the enzyme terminal deoxynucleotidyl transferase (TdT) (Ferreira and Afreen 2017). Interestingly, our TUNEL results had depicted that the quadruple-combinatorial treatment (TCNP) had significantly induced apoptosis as evident from the highest percentage of TUNEL-positive cells with brown and shrunken nuclei ( Fig.…”

Section: Discussionmentioning

confidence: 99%

Dynamics of repurposing synthetic and natural compounds against WNT/β-Catenin signaling in glioma- Anin vivoapproach

Precilla

Kuduvalli

Biswas

et al. 2022

Preprint

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Background: Glioblastomas arise from multistep tumorigenesis of the glial cells and are associated with poor prognosis. Despite the current state-of-art treatment, tumor recurrence is inevitable. Thus, there exists a desperate need for effective therapeutic alternatives to improve glioblastoma outcome. Among the innovations blooming up, drug repurposing could provide a profound premises for glioblastoma treatment enhancement. While considering this strategy, the efficacy of the repurposed drugs as monotherapies were not up to par; hence, the focus has now shifted to investigate the multi-drug combinations to target glioblastomas. In line with this concept, we investigated the efficacy of a quadruple-combinatorial treatment comprising temozolomide (the benchmark drug) along with chloroquine (a synthetic drug), naringenin (a flavonoid) and phloroglucinol (a marine derivative) in an orthotopic glioma-induced xenograft model. Methods: Anti-proliferative effect of the drugs was assessed by immunostaining. The expression profiles of WNT/β-catenin and apoptotic markers were evaluated by qRT-PCR, immunoblotting and ELISA. Patterns of mitochondrial depolarization was determined by flow cytometry. TUNEL assay was performed to affirm apoptosis induction. In vivo drug detection study was carried out by ESI-Q-TOF MS analysis. Results: The quadruple-drug treatment had significantly hampered GB proliferation and had induced apoptosis by modulating the WNT/β-catenin signalling. Flow cytometric analysis revealed that the induction of apoptosis was associated with mitochondrial depolarization. Further the quadruple-drug cocktail, had breached the blood brain barrier and was detected in the brain tissue and plasma samples from various experimental groups. Conclusion: The quadruple-drug combination served as a promising adjuvant therapy to combat glioma lethality in vivo and can be probed for translation from bench to bedside.

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Methods related to studying tau fragmentation

Cited by 6 publications

References 31 publications

Dysregulation of Exosome Cargo by Mutant Tau Expressed in Human-induced Pluripotent Stem Cell (iPSC) Neurons Revealed by Proteomics Analyses

Dysregulation of Exosome Cargo by Mutant Tau Expressed in Human-induced Pluripotent Stem Cell (iPSC) Neurons Revealed by Proteomics Analyses

Repurposing synthetic and natural derivatives induces apoptosis in an orthotopic glioma‐induced xenograft model by modulating WNT/β‐catenin signaling

Dynamics of repurposing synthetic and natural compounds against WNT/β-Catenin signaling in glioma- Anin vivoapproach

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