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Proteomic analysis of the cornified layer of the epidermis on the soles of mice carrying a keratinocyte-specific deletion of Autophagy-related gene 7 (Atg7)

ABSTRACT: Keratinocytes of the epidermis undergo massive remodeling of intracellular structures to enter the stratum corneum. This work investigates the degree to which autophagy participates in the remodeling by comparing corneocytes from the sole (foot pads) of wild type (WT, n=5, samples names NK70-74) to those from mice (EKO, n=6, samples names NK75-80) with epidermal keratinocyte-specific deletion of Autophagy-related gene 7 (Atg7). Sample NK81 is a blank. The cornified layer of the epidermis (stratum corneum) was sampled by tape-stripping from the soles of hind limbs and subjected to label-free proteomics as previously described (Karim et al. PLoS One 2023;18:e0283619). Among more than 170 proteins detected, 10 proteins were significantly increased in abundance when autophagy was suppressed. The autophagy-dependent alterations in the protein composition of corneocytes are less pronounced than those in hard cornified skin appendages. Since the proteome of the stratum corneum depends partly on autophagy in keratinocytes, it may be used for the detection of autophagy-dependent aberration of keratinocyte differentiation.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Leopold Eckhar

PROVIDER: MSV000095353 | MassIVE | Tue Jul 16 15:01:00 BST 2024

SECONDARY ACCESSION(S): PXD053990

REPOSITORIES: MassIVE

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Project description:Cursons2015 - Regulation of ERK-MAPK signaling in human epidermis Model comparing the abundance of phosphorylated MAPK signalling proteins and calcium signalling in the epidermis. This model is described in the article: Regulation of ERK-MAPK signaling in human epidermis. Cursons J, Gao J, Hurley DG, Print CG, Dunbar PR, Jacobs MD, Crampin EJ. BMC Syst Biol 2015; 9: 41 Abstract: The skin is largely comprised of keratinocytes within the interfollicular epidermis. Over approximately two weeks these cells differentiate and traverse the thickness of the skin. The stage of differentiation is therefore reflected in the positions of cells within the tissue, providing a convenient axis along which to study the signaling events that occur in situ during keratinocyte terminal differentiation, over this extended two-week timescale. The canonical ERK-MAPK signaling cascade (Raf-1, MEK-1/2 and ERK-1/2) has been implicated in controlling diverse cellular behaviors, including proliferation and differentiation. While the molecular interactions involved in signal transduction through this cascade have been well characterized in cell culture experiments, our understanding of how this sequence of events unfolds to determine cell fate within a homeostatic tissue environment has not been fully characterized.We measured the abundance of total and phosphorylated ERK-MAPK signaling proteins within interfollicular keratinocytes in transverse cross-sections of human epidermis using immunofluorescence microscopy. To investigate these data we developed a mathematical model of the signaling cascade using a normalized-Hill differential equation formalism.These data show coordinated variation in the abundance of phosphorylated ERK-MAPK components across the epidermis. Statistical analysis of these data shows that associations between phosphorylated ERK-MAPK components which correspond to canonical molecular interactions are dependent upon spatial position within the epidermis. The model demonstrates that the spatial profile of activation for ERK-MAPK signaling components across the epidermis may be maintained in a cell-autonomous fashion by an underlying spatial gradient in calcium signaling.Our data demonstrate an extended phospho-protein profile of ERK-MAPK signaling cascade components across the epidermis in situ, and statistical associations in these data indicate canonical ERK-MAPK interactions underlie this spatial profile of ERK-MAPK activation. Using mathematical modelling we have demonstrated that spatially varying calcium signaling components across the epidermis may be sufficient to maintain the spatial profile of ERK-MAPK signaling cascade components in a cell-autonomous manner. These findings may have significant implications for the wide range of cancer drugs which therapeutically target ERK-MAPK signaling components. This model is hosted on BioModels Database and identified by: BIOMD0000000659. To cite BioModels Database, please use: Chelliah V et al. BioModels: ten-year anniversary. Nucl. Acids Res. 2015, 43(Database issue):D542-8. To the extent possible under law, all copyright and related or neighbouring rights to this encoded model have been dedicated to the public domain worldwide. Please refer to CC0 Public Domain Dedication for more information.

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Proteomic analysis of the cornified layer of the epidermis on the soles of mice carrying a keratinocyte-specific deletion of Autophagy-related gene 7 (Atg7)

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