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D6_T30

ABSTRACT: To examine the signaling pathways active downstream ACKR2, both in constitutive and ligand-stimulated conditions, we carried out a large-scale mass spec-based quantitative phosphoproteomic analysis by SILAC technique. ACKR2 has been expressed in HEK293T cells using a tetracycline-inducible system and stimulated with the common ligand CCL3L1. Using computational approaches, we performed a comparative system-based analysis of the ACKR2- and CCR5-mediated phosphoproteomes.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: gabriella tedeschi

LAB HEAD: Gabriella Tedeschi

PROVIDER: PXD009851 | Pride | 2023-03-10

REPOSITORIES: Pride

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			Action	DRS
	ElautoP_1.raw	Raw
	EluatoB1_1.raw	Raw
	EluatoB1_2.raw	Raw
	EluatoB1_3.raw	Raw
	EluatoB2_1.raw	Raw

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Publications

Phosphoproteomic mapping of CCR5 and ACKR2 signaling properties.

Vacchini Alessandro A Maffioli Elisa E Di Silvestre Dario D Cancellieri Cinzia C Milanesi Samantha S Nonnis Simona S Badanai Sabrina S Mauri Pierluigi P Negri Armando A Locati Massimo M Tedeschi Gabriella G Borroni Elena Monica EM

Frontiers in molecular biosciences 20221122

ACKR2 is an atypical chemokine receptor which is structurally uncoupled from G proteins and is unable to activate signaling pathways used by conventional chemokine receptors to promote cell migration. Nonetheless, ACKR2 regulates inflammatory and immune responses by shaping chemokine gradients in tissues <i>via</i> scavenging inflammatory chemokines. To investigate the signaling pathways downstream to ACKR2, a quantitative SILAC-based phosphoproteomic analysis coupled with a systems biology appr ...[more]

PMID: 36483536

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D6_T0

Project description:To examine the signaling pathways active downstream ACKR2, both in constitutive and ligand-stimulated conditions, we carried out a large-scale mass spec-based quantitative phosphoproteomic analysis by SILAC technique. ACKR2 has been expressed in HEK293T cells using a tetracycline-inducible system and stimulated with the common ligand CCL3L1. Using computational approaches, we performed a comparative system-based analysis of the ACKR2- and CCR5-mediated phosphoproteomes.

2023-03-10 | PXD009865 | Pride

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Zebrafish embryo T18°C MAX QUANT

Project description:Effect of temperature variation on zebra fish health and behaviour Water temperature is an important environmental parameter that influences the distribution and the health of fishes. Moreover, temperature has a central role in ectothermic animals affecting their physiology and behaviour. The aim of this study was to determine the effects of ambient temperature on the molecular mechanism and the behavioural responses in ZF (Danio rerio) (ZF) a widely used animal model for environmental "omics" genomics researches. A global proteomic analysis was performed on fish brains to investigate physiological and biochemical changes that may occur from the exposure to this environmental stressor at the central nervous system level. Behavioural investigations were performed using a Y-Maze task to evaluate how temperature variations may affect swimming performance, the response to novelty and the spatial memory. Adult specimens of wild type ZF were kept at three different temperatures: 18 °C, 34 °C and 26 °C (used as a control) for 21 days. Proteomic data revealed 202XXX differentially expressed proteins across acclimation groups. These proteins are involved in energy metabolism, mitochondrial regulation and cytoskeletal organization. Temperature variation was able to alter swimming performance in adult ZF and to induce variations in the novel ambient exploration and in the acquisition and consolidation of spatial memory. This study indicates that moderate temperature variations can elicit biochemical changes that may affect fish health and behaviour. This combined approach may provide insights into mechanisms supporting thermal adaptation and plasticity in fishes.

2024-01-26 | PXD009934 | Pride

Proteome of Saccharomyces cerevisiae extracellular matrix

Project description:Proteomic analysis of the extracellular matrix of Saccharomyces cerevisiae W303-1A Wt and the isogenic mutant strain gup1Δ during the development of multicellular overlays.

2015-11-30 | PXD001133 | Pride

Microarray analysis of ApcMin/+ and D6(ACKR2)-/-ApcMin/+ tumors

Project description:Mast cells have been suggested to either promote or suppress tumor progression but the mechanisms underlying these outcomes are unclear. Here, we show that mice with a heterozygous mutation in the adenomatous polyposis coli gene (ApcMin/+) displayed reduced intestinal tumor burden and increased survival in the background of the chemokine decoy receptor ACKR2 deficiency (ACKR2-/-). The ACKR2-/-ApcMin/+ tumors showed increased infiltration of mast cells. In mast cell-deficient Sash (c-kitW-sh/W-sh) ACKR2-/-ApcMin/+ mice the survival advantage was lost as the tumors grew rapidly and adoptive transfer of mast cells reduced the tumor burden in these mice. The tumor burden is also increased in Rag2-/-ACKR2-/-ApcMin/+ mice and the protection is reconstituted by adoptively transferred CD8+ T-lymphocytes. Mast cells from ACKR2-/- mice expressed elevated levels of chemokine receptors CCR2 and CCR5 and are also efficient in antigen presentation and activation of CD8+ T-cells. Mast cell-derived leukotriene B4 is a critical mediator of CD8+ T-lymphocyte recruitment as the BLT1-/-ACKR2-/-ApcMin/+ mice are highly susceptible to intestinal tumor induced mortality. Taken together, these data demonstrate that chemokine mediated mast cell recruitment is essential for initiating LTB4/BLT1 regulated CD8+ T-cell homing and generation of effective anti-tumor immunity against intestinal tumor development.

2017-12-31 | GSE104829 | GEO

Microarray analysis of Bone marrow derived mast cells (BMMCs) from C57BL/6 and ACKR2-/- mice

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Chemoradiotherapy-induced ACKR2+ tumor cells drive CD8+ T cell senescence and tumor recurrence

Project description:T cell senescence is a dysfunctional state distinct from exhaustion, yet the mechanism driving CD8+ T cell senescence directly in tumor microenvironment and its impact on cancer prognosis remain elusive. Here, by using single cell RNA sequencing of human cervical cancer tissues, we found that concurrent chemoradiotherapy (CCRT) specifically promoted CD8+ T cell senescence, which is driven by a subpopulation of ACKR2+ CCRT-resistance tumor cells. Mechanistically, in response to CCRT treatment, ACKR2 was directly induced and could also be upregulated through its ligands CC chemokines that produced by activated myeloid and T cells. Subsequently, these ACKR2+ tumor cell subpopulation also produced TGFβ to drive CD8+ T cell senescence, thereby compromising antitumor immunity. Moreover, retrospective analysis showed that ACKR2 level and CD8+ T cell senescence was greatly enhanced in cervical cancer patients who recurred after CCRT, thereby predicting poor prognosis in these patients. Overall, we identified a subpopulation of CCRT-resistant ACKR2+ tumor cells in driving CD8+ T cell senescence and thus tumor recurrence, which may represent a critical mechanism in mediating CCRT resistance, and highlighted the therapeutic potential of targeting ACKR2 or CD8+ T cell senescence in the treatment of CCRT-resistant cancer patients.

2024-06-07 | GSE236738 | GEO

A protein phosphatase network controls the temporal and spatial dynamics of differentiation commitment in human epidermis

Project description:Epidermal homeostasis depends on a balance between stem cell renewal and terminal differentiation. The transition between the two cell states,termed commitment, is poorly understood. Here we characterise commitment by integrating transcriptomic and proteomic data from disaggregated primary human keratinocytes held in suspension to induce differentiation. Cell detachment induces several protein phosphatases, five of which - DUSP6, PPTC7, PTPN1, PTPN13 and PPP3CA – promote differentiation by negatively regulating ERK MAPK and positively regulating AP1 transcription factors. Conversely, DUSP10 expression antagonises commitment. The phosphatases form a dynamic network of transient positive and negative interactions that change over time, with DUSP6 predominating at commitment. Boolean network modelling identifies a mandatory switch between two stable states (stem and differentiated) via an unstable (committed) state. Phosphatase expression is also spatially regulated in vivo and in vitro. We conclude that an auto-regulatory phosphatase network maintains epidermal homeostasis by controlling the onset and duration of commitment.

2017-10-18 | PXD003281 | Pride

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