Dataset Information

Proteomic analysis of renal cancer secretome

ABSTRACT: Introduction/aim: Renal cell cancer (RCC) is the most frequent kidney malignancy. Here, we aimed to analyze RCC secretome to find key molecules involved in tumor microenvironment regulation. Methods: Conditioned media from five RCC-derived cell lines (Caki-1, KIJ-265T, KIJ-308T, 786-O, and A498) and a non-cancerous proximal tubules (RPTEC) were used for mass spectrometry based proteomic analysis. The results were evaluated using Gene Onthology algorithms. Results: When compared with RPTEC, CM of Caki-1 and KIJ265T contained 587 and 802 aberrantly expressed proteins, respectively. Remarkably, there were 360 proteins commonly found in CM from Caki-1 and KIJ265T cells, with striking 348 proteins with changes in similar directions. 786-O CM expressed 612, A498 569, and KIJ308T 802 proteins. All cell lines shared 124 aberrantly expressed CM proteins, with 119 proteins with changes in the same direction. Top commonly downregulated proteins included SCIN, CBR4, MUC1, PELP1, LSM1, while top upregulated proteins were BZW2, COLGALT1, EEF1A2, CSDE1, INHBA, and STAT1. GO analysis of cellular components related to the identified proteins revealed that 63 proteins were associated with extracellular exosomes. Conclusion: Different RCC cell lines secrete proteins, commonly altered when compared with normal kidney proximal tubules. 53% of these proteins are associated with extracellular exosomes.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Agnieszka Piekielko-Witkowska

PROVIDER: MSV000088490 | MassIVE | Thu Dec 02 03:53:00 GMT 2021

SECONDARY ACCESSION(S): PXD030085

REPOSITORIES: MassIVE

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Publications

Coordinated reprogramming of renal cancer transcriptome, metabolome and secretome associates with immune tumor infiltration.

Poplawski Piotr P Alseekh Saleh S Jankowska Urszula U Skupien-Rabian Bozena B Iwanicka-Nowicka Roksana R Kossowska Helena H Fogtman Anna A Rybicka Beata B Bogusławska Joanna J Adamiok-Ostrowska Anna A Hanusek Karolina K Hanusek Jan J Koblowska Marta M Fernie Alisdair R AR Piekiełko-Witkowska Agnieszka A

Cancer cell international 20230105 1

<h4>Background</h4>Clear cell renal cell carcinoma (ccRCC) is the most common subtype of renal cancer. The molecules (proteins, metabolites) secreted by tumors affect their extracellular milieu to support cancer progression. If secreted in amounts detectable in plasma, these molecules can also serve as useful, minimal invasive biomarkers. The knowledge of ccRCC tumor microenvironment is fragmentary. In particular, the links between ccRCC transcriptome and the composition of extracellular milieu ...[more]

PMID: 36604669

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Project description:Background: Acute kidney injury (AKI) is a common clinical event with high morbidity and mortality. We previously demonstrated that injection of cord blood endothelial colony forming cell (ECFC)-derived exosomes, highly enriched in miRNA(miR)-486-5p, prevented ischemic kidney injury in mice. While preclinical models support involvement of several miRs in AKI, the direct effects of miR-486-5p on injury and the kidney transcriptome are unknown. Objective: We studied effects of miR-486-5p in mice with kidney ischemia-reperfusion (IR) injury, and compared the impact of miR-486-5p and ECFC-derived exosomes on the transcriptome of proximal tubules (PTs) and renal endothelial cells. Methods: Adult male mice were subjected to bilateral kidney ischemia (30 min), and injected via tail vein with or without vehicle, miR-486-5p mimic, ECFC-derived exosomes, or scramble miR at the start of reperfusion. Plasma and tissues were collected 24 hrs after reperfusion, and proximal tubular and endothelial cells were isolated for mRNA analysis by RNA-Seq. Results: In mice with kidney IR, injection of miR-486-5p mimic increased levels of miR-486-5p in proximal tubules and endothelial cells, and significantly improved plasma creatinine, histological injury, neutrophil infiltration, and apoptosis, compared to vehicle treatment. MiR-486-5p inhibited expression of phosphatase and tensin homolog (PTEN), and activated AKT. Similar results were observed with exosomes, but scramble miR had no effect. In proximal tubules, miR-486-5p or exosomes reduced expression of several AKI genes (e.g. Lcn2, Havcr1 and Krt20) and caused alterations in apoptotic genes compared to IR alone. In endothelium, miR-486-5p or exosomes caused milder effect than in PTs but still impacting on expression of injury-related genes. Conclusion: MiR-486-5p protects mice against ischemic kidney injury. Both miR-486-5p and exosomes reduce expression of apoptotic genes in PTs and endothelium. The results suggest that systemic delivery of miR-486-5p has therapeutic potential in ischemic AKI.

Project description:BACKGROUND: Ischemia/reperfusion injury (IRI) is the leading cause of acute kidney injury (AKI). The current standard of care focuses on supporting kidney function, stating the need for more efficient and targeted therapies to enhance repair. Mesenchymal Stromal Cells (MSCs) and their secretome, either as conditioned medium (CM) or extracellular vesicles (EVs), have emerged as promising options for regenerative therapy, however, their full potential in treating AKI remains unknown.METHODS: In this study, we employed an in vitro model of chemically-induced ischemia using antimycin A combined with 2-deoxy-D-glucose to induce ischemic injury in proximal tubule epithelial cells. Afterwards, we evaluated the effects of MSC secretome, CM or EVs obtained from adipose tissue, bone marrow and umbilical cord, on ameliorating the detrimental effects of ischemia. To assess the damage and treatment outcomes, we analyzed cell morphology, mitochondrial health parameters (mitochondrial activity, ATP production, mass and membrane potential) and overall cell metabolism by metabolomics.RESULTS: Our findings show that ischemic injury caused cytoskeletal changes confirmed by disruption of the F-actin network, energetic imbalance as revealed by a 50% decrease in the oxygen consumption rate, increased oxidative stress, mitochondrial dysfunction and reduced cell metabolism. Upon treatment with MSC secretome, the morphological derangements were partly restored and ATP production increased by 40-50%, with umbilical cord-derived EVs being most effective. Furthermore, MSC treatment led to phenotype restoration as indicated by an increase in cell bioenergetics, including increased levels of glycolysis intermediates, as well as an accumulation of antioxidant metabolites.CONCLUSION: Our in vitro model effectively replicated the in vivo-like morphological and molecular changes observed during ischemic injury. Additionally, treatment with MSC secretome ameliorated proximal tubule damage, highlighting its potential as a viable therapeutic option for targeting AKI.

Dataset Information

Proteomic analysis of renal cancer secretome

Publications

Coordinated reprogramming of renal cancer transcriptome, metabolome and secretome associates with immune tumor infiltration.

Similar Datasets

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