Dataset Information

Proteomic Dissection of Nanotopography-Sensitive Mechanotransductive Signaling Hubs that Foster Neuronal Differentiation in PC12 Cells.

ABSTRACT: Neuronal cells are competent in precisely sensing nanotopographical features of their microenvironment. The perceived microenvironmental information will be "interpreted" by mechanotransductive processes and impacts on neuronal functioning and differentiation. Attempts to influence neuronal differentiation by engineering substrates that mimic appropriate extracellular matrix (ECM) topographies are hampered by the fact that profound details of mechanosensing/-transduction complexity remain elusive. Introducing omics methods into these biomaterial approaches has the potential to provide a deeper insight into the molecular processes and signaling cascades underlying mechanosensing/-transduction but their exigence in cellular material is often opposed by technical limitations of major substrate top-down fabrication methods. Supersonic cluster beam deposition (SCBD) allows instead the bottom-up fabrication of nanostructured substrates over large areas characterized by a quantitatively controllable ECM-like nanoroughness that has been recently shown to foster neuron differentiation and maturation. Exploiting this capacity of SCBD, we challenged mechanosensing/-transduction and differentiative behavior of neuron-like PC12 cells with diverse nanotopographies and/or changes of their biomechanical status, and analyzed their phosphoproteomic profiles in these settings. Versatile proteins that can be associated to significant processes along the mechanotransductive signal sequence, i.e., cell/cell interaction, glycocalyx and ECM, membrane/f-actin linkage and integrin activation, cell/substrate interaction, integrin adhesion complex, actomyosin organization/cellular mechanics, nuclear organization, and transcriptional regulation, were affected. The phosphoproteomic data suggested furthermore an involvement of ILK, mTOR, Wnt, and calcium signaling in these nanotopography- and/or cell mechanics-related processes. Altogether, potential nanotopography-sensitive mechanotransductive signaling hubs participating in neuronal differentiation were dissected.

SUBMITTER: Maffioli E

PROVIDER: S-EPMC5758595 | biostudies-literature | 2017

REPOSITORIES: biostudies-literature

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Publications

Proteomic Dissection of Nanotopography-Sensitive Mechanotransductive Signaling Hubs that Foster Neuronal Differentiation in PC12 Cells.

Maffioli Elisa E Schulte Carsten C Nonnis Simona S Grassi Scalvini Francesca F Piazzoni Claudio C Lenardi Cristina C Negri Armando A Milani Paolo P Tedeschi Gabriella G

Frontiers in cellular neuroscience 20180104

Neuronal cells are competent in precisely sensing nanotopographical features of their microenvironment. The perceived microenvironmental information will be "interpreted" by mechanotransductive processes and impacts on neuronal functioning and differentiation. Attempts to influence neuronal differentiation by engineering substrates that mimic appropriate extracellular matrix (ECM) topographies are hampered by the fact that profound details of mechanosensing/-transduction complexity remain elusiv ...[more]

PMID: 29354032

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Project description:Neuronal cells are competent in precisely sensing nanotopographical features of their microenvironment and the intrinsic information impacts on neuronal functioning and differentiation “interpreted” by mechanotransductive processes. Attempts to influence and direct neuronal differentiation behaviour by engineering substrates that mimic appropriate extracellular matrix (ECM) topographies are hampered by the difficulty since many details of mechanosensing/-transduction are still elusive. Introducing omics methods into these biomaterial approaches has the potential to provide a deeper insight into the molecular processes and signalling cascades underlying mechanosensing/-transduction but their exigence in cellular material is often opposed by technical limitations of the biomaterial fabrication methods. Supersonic cluster beam deposition (SCBD) of zirconia nanoparticles allows instead the realisation of large macroscopic areas with biocompatible nanostructured films equipped with controllable and reproducible ECM-like nanoroughness that have been recently shown to foster neuron differentiation and maturation. Exploiting this capacity of SCBD, we challenged mechanosensing/-transduction and differentiative behaviour of neuron-like PC12 cells interacting with diverse nanotopographies and/or by changing their biomechanical status, to analyse their phosphoproteomic profiles in these settings. Versatile proteins were found to be affected that can be associated to significant processes along the mechanotransductive signal sequence, i.e. cell/cell interaction, glycocalyx and ECM, membrane/f-actin linkage and integrin activation, cell/substrate interaction, IAC, actomyosin organisation/cellular mechanics, nuclear organisation and transcription. The phosphoproteomic data suggested furthermore an involvement of ILK, mTor, Wnt and calcium signalling in these nanotopography- and/or cell mechanics-related processes. Altogether, potential nanotopography-sensitive, mechanotransductive signalling hubs participating to neuronal differentiation were dissected.

Project description:Stem cell differentiation can be regulated by biophysical cues such as nanotopography. It involves sensing and integration of these biophysical cues into their transcriptome with a mechanism that is yet to be discovered. In addition to the cytoskeletal and focal adhesion remodeling, nanotopography has also been shown to modulate nucleus morphology. Here, we studied the effect of nanotopography on the temporal changes in nuclei of human embryonic stem cells (hESCs) and human mesenchymal stem cells (hMSCs). Using a high throughput Multi-architecture (MARC) chip analysis, the circularity of the stem cell nuclei changed significantly on different patterns. Human ESCs and MSCs showed different temporal changes in nucleus morphology, lamin A/C expression and histone methylation during topography-induced neuronal differentiation. In hESCs, the expression of nuclear matrix protein, lamin A/C during neuronal differentiation of hESCs on PDMS samples were weakly detected in the first 7 days of differentiation. The histone 3 trimethylation on Lysine 9 (H3K9me3) decreased after differentiation initiated and showed temporal changes in their expression and organization during neuronal differentiation. In hMSCs, the expression of lamin A/C was significantly increased after the first 24 h of cell culture. The quantitative analysis of histone methylation also showed a significant increase in hMSCs histone methylation on 250 nm anisotropic nanogratings within the first 24 h of seeding. This reiterates the importance of cell-substrate sensing within the first 24 h for adult stem cells. The lamin A/C expression and histone methylation shows a correlation of epigenetic changes in early events of differentiation, giving an insight on how extracellular nanotopographical cues are transduced into nuclear biochemical signals. Collectively, these results provide more understanding into the nuclear regulation of the mechanotransduction of nanotopographical cues in stem cell differentiation.

Dataset Information

Proteomic Dissection of Nanotopography-Sensitive Mechanotransductive Signaling Hubs that Foster Neuronal Differentiation in PC12 Cells.

Publications

Proteomic Dissection of Nanotopography-Sensitive Mechanotransductive Signaling Hubs that Foster Neuronal Differentiation in PC12 Cells.

Similar Datasets

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