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Proteomic analysis of reticulocytes treated with the PIEZO-1 activator YODA

ABSTRACT: We performed a label free proteomic analysis on reticulocytes produced in vitro from CD34+ of healthy donors cells either exposed to DMSO as a control or to the PIEZO1 activator YODA1 at 1µM.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Cell, Blood

SUBMITTER: Virginie Salnot

LAB HEAD: Loïc Garçon

PROVIDER: PXD035122 | Pride | 2023-03-11

REPOSITORIES: Pride

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			Action	DRS
	LoGA201006__RET_1D_nvx_SCX1_F39064.raw	Raw
	LoGA201006__RET_1D_nvx_SCX2_F39065.raw	Raw
	LoGA201006__RET_1D_nvx_SCX3_F39066.raw	Raw
	LoGA201006__RET_1D_nvx_SCX4_F39067.raw	Raw
	LoGA201006__RET_1D_nvx_SCX5_F39068.raw	Raw

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Publications

Red blood cell proteomics reveal remnant protein biosynthesis and folding pathways in PIEZO1-related hereditary xerocytosis.

Caulier Alexis A Jankovsky Nicolas N Gautier Emilie Fleur EF El Nemer Wassim W Guitton Corinne C Ouled-Haddou Hakim H Guillonneau François F Mayeux Patrick P Salnot Virginie V Bruce Johanna J Picard Véronique V Garçon Loïc L

Frontiers in physiology 20221201

Hereditary xerocytosis is a dominant red cell membrane disorder characterized by an increased leak of potassium from the inside to outside the red blood cell membrane, associated with loss of water leading to red cell dehydration and chronic hemolysis. 90% of cases are related to heterozygous gain of function mutations in PIEZO1, encoding a mechanotransductor that translates a mechanical stimulus into a biological signaling. Data are still required to understand better PIEZO1-HX pathophysiology. ...[more]

PMID: 36531183

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Project description:Proteome of purified erythrocytes from blood samples from patients diagnosed with a PIEZO1-HX based on familial history, non-spherocytic chronic hemolysis, typical osmotic gradient ektacytometry and molecular testing and from healthy donors were analyzed by a label free quantification approach.

2023-03-11 | PXD031963 | Pride

Investigating the proteome of red blood cells and reticulocytes in Polycythemia Vera and Essential Thrombocythemia

Project description:The project aims at comparing the effect of the JAK2V617F mutation on protein expression in human erythroid cells in the context of Polycythemia Vera and Essential Thrombocythemia

2022-02-17 | PXD029965 | Pride

Reticulocytes vs mature red blood cell

Project description:comparative study reticulocytes vs mature red blood cell

2017-11-09 | PXD003851 | Pride

Absolute proteome quantification of highly purified reticulocytes and erythrocytes

Project description:Erythrocytes are the most numerous cells of the human body (Sender 2016). Their lifespan in the blood stream is around 120 days. The turnover implies the production of 2 million new reticulocytes each second. Circulating reticulocytes achieve their maturation in 2 to 3 days. During this short time, important modifications occur such as membrane remodeling, removing of remaining organelles and specific elimination of useless proteins such as the transferrin receptor. This remodeling will ensure the optimization of the erythrocyte ability to fulfil its transport functions. We performed a deep label-free quantitative proteome analysis of purified erythrocytes and reticulocytes to better understand the physiology of the erythrocytes and the last steps of their maturation.

2018-09-18 | PXD009258 | Pride

Proteomic characterization of exosomes derived from malaria-infected reticulocytes, in a murine malaria model

Project description:Reticulocyte-derived exosomes (rex) are 30-100 nm membrane vesicles of endocytic origin released during the maturation of reticulocytes to erythrocytes upon fusion of multivesicular bodies with the plasma membrane. We previously found that immunization of mice with rex from BALB/c mice infected with the reticulocyte-prone non-lethal P. yoelii 17X malaria strain (rexPy) in combination with CpG-ODN promoted survival and long lasting protection of mice subsequently challenged with a lethal strain. Here, we show that rexPy-mediated protection is completely lost in splenectomized animals and such protection is achieved after passive transfer of splenocytes obtained from animals immunized with rexPy+CpG. Notably, rexPy immunization of mice induced non-exhausted memory T cell expansion with effector phenotype. Proteomics analysis of rexPy confirmed their reticulocyte origin and demonstrated the presence of parasite antigens. Our studies thus prove, for what we believe is the first time, that rex from reticulocyte-prone malarial infections are able to induce non-exhausted splenic long-lasting memory responses. In order to translate these results to humans, in vitro experiments with spleen cells of human transplantation donors were performed. Human splenocytes were able to actively capture plasma45 derived exosomes from patients infected with Plasmodium vivax (exPv) and stimulation of spleen cells with exPv for 72h lead to an increase in the number of T cells. All together, these data further support the value of reticulocyte-derived exosomes as a potential novel vaccine and platform against malaria.

2018-10-26 | PXD005048 | Pride

Glyoxal induces senescence of human keratinocytes through oxidative stress and activation of the AKT/FOXO3a/p27KIP1 pathway

Project description:Senescence is a well-known cellular event characterized by specific markers like a permanent cell proliferation arrest and the secretion of messenger molecules forming the Senescence-Associated Secretory Phenotype (SASP). The SASP composition depends on many factors such as the cell type or the nature of the stress that induced senescence. Since the skin constitutes a barrier with the external environment, it is particularly subjected to different types of stresses and thus to premature cellular aging. The dicarbonyl compounds glyoxal and methylglyoxal are precursors of Advanced Glycation End-products (AGEs), known to be a feature of normal and pathological aging. In this study, we have demonstrated that glyoxal provokes oxidative stress by increasing ROS and AGEs levels and can induce senescence in human keratinocytes. An “early-stage” senescence phenotype characterized by a cell cycle arrest mediated by the AKT-FOXO3-p27KIP1 pathway and a “late-stage” senescence maintained by the p16INK4/pRb pathway were evidenced. Moreover, we characterized the resulting secretory phenotype during early senesecence by mass spectrometry. Our study provides evidence that glyoxal can affect keratinocyte function and act as a driver of human skin aging. Hence, senotherapeutics aimed at modulating glyoxal-associated senescence phenotype could be relevant to prevent the aging process in skin.

2022-01-03 | PXD025184 | Pride

Multiparametric characterization of red blood cells physiology after hypotonic dialysis based drug encapsulation process.

Project description:Red blood cells (RBC) can act as carriers for therapeutic agents and given their biocompatibility and long lifespan in the circulation, they can substantially improve the safety, pharmacokinetics, and pharmacodynamics of many drugs. Maintaining RBC integrity and lifespan is important for the efficacy of RBC as drug carrier and can be a complex challenge of drug encapsulation. We investigated the impact of drug encapsulation by hypotonic dialysis on RBC physiology and integrity. Several parameters were compared between processed RBC loaded with L-asparaginase (Ã¢â‚¬Å“eryaspaseÃ¢â‚¬ï¿½), processed RBC without drug and non-processed RBC. Processed RBC were less hydrated and displayed a reduction of intracellular content, relative to non-processed RBC. This reduction of intracellular content changed the RBC metabolomic profile, as indicated by the activation of the pentose phosphate pathway, but no impact on the global RBC proteomic profile was observed. We found that the encapsulation process caused moderate morphological changes and was accompanied by an increase of microparticles. Despite a decrease in their deformability, processed RBC were not mechanically retained in a spleen-mimicking device. Moreover, processed RBC had increased surface-to-volume ratio and osmotic resistance. Finally, the half-life of processed RBC was not significantly affected in a mouse model and our previous phase 1 clinical study showed that encapsulation of asparaginase in RBC prolonged its in vivo half-life compared to free form. Overall, our study demonstrated that encapsulation by hypotonic dialysis may affect certainÃ‚Â several characteristics of RBC, but does not significantly impact on the in vivo longevity of RBC or their drug carrier function.

2021-11-29 | PXD022549 | Pride

RNA-seq of wildtype and PIEZO1 knockout mice livers to investigate lipid and glucose regulation by the endothelial force sensor PIEZO1

Project description:PIEZO1 is a mechanically-activated ion channel that contributes to flow sensing in vascular endothelium. Moreover, deletion of endothelial PIEZO1 was recently found to suppress activation of Notch1 target genes in hepatic microvascular endothelium. Here, because of the liver’s dominant role in lipid regulation, we set out to test the novel hypothesis that endothelial PIEZO1 regulates hepatic lipid homeostasis. We performed bulk RNA sequencing on PIEZO1-deleted mice exposed to chow and high fat diets. Our transcriptomics analysis reveal unexpected relevance to lipid and glucose homeostasis.

2024-06-29 | E-MTAB-9996 | biostudies-arrayexpress

Dynamic profiling of the protein life cycle in response to pathogens (protein)

Project description:Protein expression is regulated by production and degradation of mRNAs and proteins, but their specific relationships remain unknown. We combine measurements of protein production and degradation and mRNA dynamics to build a quantitative genomic model of the differential regulation of gene expression in LPS stimulated mouse dendritic cells. Changes in mRNA abundance play a dominant role in determining most dynamic fold changes in protein levels. Conversely, the preexisting proteome of proteins performing basic cellular functions is remodeled primarily through changes in protein production or degradation, accounting for over half of the absolute change in protein molecules in the cell. Thus, the proteome is regulated by transcriptional induction of novel cellular functions and remodeling of preexisting functions through the protein life cycle. Mouse primary dendritic cells were treated with LPS or mock stimulus and profiled over a 12-hour time course. Cells were grown in M-labeled SILAC media, which was replaced with H-labeled SILAC media at time 0. Aliquots were taken at 0, 0.5, 1, 2, 3, 4, 5, 6, 9, and 12 hours post-stimulation and added to equal volumes of a master mix of unlabeled (L) cells for the purpose of normalization. RNA-Seq was performed at 0, 1, 2, 4, 6, 9, and 12 hours post-stimulation.

2015-03-06 | E-GEOD-59792 | biostudies-arrayexpress

Mechanical regulation of the electric maturation of hippocampal neurons

Project description:In this project, we cultured rat hippocampal neurons in mechanically different environments and studied electrical maturation. We compared WT neurons with two different Piezo1 knockdown conditions. These two knockdown conditions were generated in two independent CRISPR-Cas9 KD assays. Each assay is based on four different CRISPR-Cas9 guides targeting the Piezo1 gene. RNA sequencing was used to analyse the pathway leading to the stiffness dependent maturation behaviour.

2024-07-10 | E-MTAB-13503 | biostudies-arrayexpress