Quantitative proteomic analysis identifies proteins and pathways related to neuronal development in differentiated SH-SY5Y neuroblastoma cells

Murillo, Jimmy Rodriguez; Goto‐Silva, Livia; Sánchez, Aniel; Nogueira, Fábio César Sousa; Domont, Gilberto B.; Junqueira, Magno

doi:10.1016/j.euprot.2017.06.001

Cited by 55 publications

(46 citation statements)

References 67 publications

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“…GO analysis of these clusters reveals that they contain proteins involved in intracellular metabolism and homeostasis, genomic translation, the cell cycle, and biosynthesis of amino acids and peptides ( Figure 2E ). Downregulation of DNA replication and cell cycle-related proteins is also reported to coincide with terminal differentiation in neuroblastoma cells and with development of cultured rat neurons ( Murillo et al, 2017 ; Frese et al, 2017 ). These results suggest that the overall proteome dynamics are indicative of cellular processes such as cell cycle exit and neuronal differentiation.…”

Section: Resultsmentioning

confidence: 97%

Quantitative mapping of transcriptome and proteome dynamics during polarization of human iPSC-derived neurons

Lindhout

Kooistra

Portegies

et al. 2020

eLife

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The differentiation of neuronal stem cells into polarized neurons is a well-coordinated process which has mostly been studied in classical non-human model systems, but to what extent these findings are recapitulated in human neurons remains unclear. To study neuronal polarization in human neurons, we cultured hiPSC-derived neurons, characterized early developmental stages, measured electrophysiological responses, and systematically profiled transcriptomic and proteomic dynamics during these steps. The neuron transcriptome and proteome shows extensive remodeling, with differential expression profiles of ~1100 transcripts and ~2200 proteins during neuronal differentiation and polarization. We also identified a distinct axon developmental stage marked by the relocation of axon initial segment proteins and increased microtubule remodeling from the distal (stage 3a) to the proximal (stage 3b) axon. This developmental transition coincides with action potential maturation. Our comprehensive characterization and quantitative map of transcriptome and proteome dynamics provides a solid framework for studying polarization in human neurons.

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Section: Resultsmentioning

confidence: 97%

Quantitative mapping of transcriptome and proteome dynamics during polarization of human iPSC-derived neurons

Lindhout

Kooistra

Portegies

et al. 2020

eLife

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“…Since there are several studies using this cell line as an in vitro model for neurodegenerative disorders (Liu et al, 2018;Murillo et al, 2017;Xicoy, Wieringa, & Martens, 2017), particularly for investigation of Aβ toxicity and drug screening (Jämsä, Belda, Edlund, & Lindström, 2011;Lattanzio, Carboni, Carretta, Candeletti, & Romualdi, 2016;Yuan et al, 2017), our first objective was to compare the effect of the peptide on undifferentiated and differentiated cells. SH-SY5Y are neuroblast-like cells that can be differentiated into a mature neuronal phenotype using several differentiating agents, including RA, phorbol esters, and neurotrophins (Kovalevich & Langford, 2013).…”

Section: Discussionmentioning

confidence: 99%

Genistein protects against amyloid‐beta‐induced toxicity in SH‐SY5Y cells by regulation of Akt and Tau phosphorylation

Petry

Coelho

Gaelzer

et al. 2019

Phytotherapy Research

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Alzheimer's disease is a neurodegenerative disorder characterized by extracellular deposition of amyloid‐β (Aβ) peptide and hyperphosphorylation of Tau protein, which ultimately leads to the formation of intracellular neurofibrillary tangles and cell death. Increasing evidence indicates that genistein, a soy isoflavone, has neuroprotective effects against Aβ‐induced toxicity. However, the molecular mechanisms involved in its neuroprotection are not well understood. In this study, we have established a neuronal damage model using retinoic‐acid differentiated SH‐SY5Y cells treated with different concentrations of Aβ25–35 to investigate the effect of genistein against Aβ‐induced cell death and the possible involvement of protein kinase B (PKB, also termed Akt), glycogen synthase kinase 3β (GSK‐3β), and Tau as an underlying mechanism to this neuroprotection. Differentiated SH‐SY5Y cells were pre‐treated for 24 hr with genistein (1 and 10 nM) and exposed to Aβ25–35 (25 μM), and we found that genistein partially inhibited Aβ induced cell death, primarily apoptosis. Furthermore, the protective effect of genistein was associated with the inhibition of Aβ‐induced Akt inactivation and Tau hyperphosphorylation. These findings reinforce the neuroprotective effects of genistein against Aβ toxicity and provide evidence that its mechanism may involve regulation of Akt and Tau proteins.

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“…GO analysis of these clusters reveals that they contain proteins involved in intracellular metabolism and homeostasis, genomic translation, the cell cycle, and biosynthesis of amino acids and peptides (FIG 2E). Downregulation of DNA replication and cell cycle-related proteins is also reported to coincide with terminal differentiation in neuroblastoma cells and with development of cultured rat neurons (Murillo et al 2017;Frese et al 2017). These results suggest that the overall proteome dynamics are indicative of cellular processes such as cell cycle exit and neuronal differentiation.…”

Section: Proteomic Profiling Of Developing Human Ipsc-derived Neuronsmentioning

confidence: 98%

Quantitative mapping of transcriptome and proteome dynamics during polarization of human iPSC-derived neurons

Lindhout

Kooistra

Portegies

et al. 2020

Preprint

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Early neuronal development is a well-coordinated process in which neuronal stem cells differentiate into polarized neurons. This process has been well studied in classical non-human model systems, but to what extent this is recapitulated in human neurons remains unclear. To study neuronal polarization in human neurons, we cultured human iPSC-derived neurons, characterized early developmental stages, measured electrophysiological responses, and systematically profiled transcriptomic and proteomic dynamics during these steps. We found extensive remodeling of the neuron transcriptome and proteome, with altered mRNA expression of ~1,100 genes and different expression profiles of ~1,500 proteins during neuronal differentiation and polarization. We also identified a distinct stage in axon development marked by an increase in microtubule remodeling and apparent relocation of the axon initial segment from the distal to proximal axon. Our comprehensive characterization and quantitative map of transcriptome and proteome dynamics provides a solid framework for studying polarization in human neurons.(FOM, #16NEPH05, CJW). AUTHOR CONTRIBUTIONSFWL, RK and SP designed the research, conducted experiments, and wrote the article together with LJH. RS conducted and analyzed the mass spectrometry experiments supervised by MA, LJH performed and interpreted the electrophysiology data supervised by CJW, RK and BLS provided and interpreted the RNA sequencing analysis. HDM edited the article. CCH designed the experimental plan, supervised the research and wrote the article.

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Quantitative proteomic analysis identifies proteins and pathways related to neuronal development in differentiated SH-SY5Y neuroblastoma cells

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Cited by 55 publications

References 67 publications

Quantitative mapping of transcriptome and proteome dynamics during polarization of human iPSC-derived neurons

Quantitative mapping of transcriptome and proteome dynamics during polarization of human iPSC-derived neurons

Genistein protects against amyloid‐beta‐induced toxicity in SH‐SY5Y cells by regulation of Akt and Tau phosphorylation

Quantitative mapping of transcriptome and proteome dynamics during polarization of human iPSC-derived neurons

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