Differential proteomics argues against a general role for CD9, CD81 or CD63 in the sorting of proteins into extracellular vesicles

Journal of Extracellular Vesicles, Год журнала: 2023, Номер 12(8)

Опубликована: Июль 31, 2023

The tetraspanins CD9, CD81 and CD63 are major components of extracellular vesicles (EVs). Yet, their impact on EV composition remains under-investigated. In the MCF7 breast cancer cell line was as expected predominantly intracellular. contrast CD9 strongly colocalized at plasma membrane, albeit with different ratios sites, which may explain a higher enrichment in EVs. Absence these had little protein analysed by quantitative mass spectrometry. We also effect concomitant knock-out because two play similar roles several cellular processes associate directly Ig domain proteins, CD9P-1/EWI-F/PTGFRN EWI-2/IGSF8. These were sole proteins significantly decreased EVs double CD9- CD81-deficient cells. case EWI-2, this is primarily consequence expression level. conclusion, study shows that CD63, commonly used markers, marginal role determining released cells highlights regulation level and/or trafficking CD9P-1 EWI-2 CD81.

Язык: Английский

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Comparison of immunotherapy mediated by apoptotic bodies, microvesicles and exosomes: apoptotic bodies’ unique anti-inflammatory potential

Journal of Translational Medicine, Год журнала: 2023, Номер 21(1)

Опубликована: Июль 17, 2023

Abstract Immunotherapy, including immunostimulation and immunosuppression, has seen significant development in the last 10 years. Immunostimulation been verified as effective anti-cancer treatment, while immunosuppression is used treatment of autoimmune disease inflammation. Currently, with update newly-invented simplified isolation methods findings potent triggered immune responses, extracellular vesicle-based immunotherapy very eye-catching. However, research on three main types vesicles, exosomes, microvesicles apoptotic bodies, needs to be more balanced. These subtypes share a certain level similarity, at same time, they have their own properties caused by different biogensis. Herein, we summarized respectively status based each kind vesicle discuss possible involved mechanisms. In conclusion, highlighted that effect body clear strong. Apoptotic bodies an excellent potential immunosuppressive anti-inflammatory therapies .

Язык: Английский

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Autophagy-Dependent Secretion: Crosstalk between Autophagy and Exosome Biogenesis

Current Issues in Molecular Biology, Год журнала: 2024, Номер 46(3), С. 2209 - 2235

Опубликована: Март 8, 2024

The cellular secretome is pivotal in mediating intercellular communication and coordinating responses to stressors. Exosomes, initially recognized for their role waste disposal, have now emerged as key messengers with significant therapeutic diagnostic potential. Similarly, autophagy has transcended its traditional a removal mechanism, emerging regulator of intracellular pathways contributor unique autophagy-dependent secretome. Secretory authophagy, initiated by various stress stimuli, prompts the selective release proteins implicated inflammation, including leaderless that bypass conventional endoplasmic reticulum–Golgi secretory pathway. This reflects impact stress-induced on secretion profiles, modulation exosome release. convergence biogenesis exemplified formation amphisomes, vesicles integrate autophagic endosomal pathways, indicating synergistic interplay. Regulatory common both particularly mTORC1, emerge potential targets alter profiles involved diseases. review explores dynamic interplay between formation, highlighting influence composition. While cytokine preconditioning well-established regenerative medicine, strategic manipulation still underexplored, presenting promising but uncharted landscape.

Язык: Английский

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Tetraspanin CD9 alters cellular trafficking and endocytosis of tetraspanin CD63, affecting CD63 packaging into small extracellular vesicles

Journal of Biological Chemistry, Год журнала: 2025, Номер unknown, С. 108255 - 108255

Опубликована: Фев. 1, 2025

Small extracellular vesicles (sEVs) are particles secreted from cells that play vital roles both in normal physiology and human disease. sEVs highly enriched tetraspanin proteins, such as CD9 CD63, contain tetraspanin-enriched membrane microdomains involved loading of with macromolecule cargoes sEV biogenesis. However, the precise individual tetraspanins biogenesis cargo remain poorly understood. Here, we report negatively regulated CD63 trafficking to its subsequent packaging into small EVs, whereas had no discernable effect on localization or packaging. Using super resolution microscopy vesicles, showed governs fraction loaded CD63. Interestingly, CD9-dependent suppression was rescued by pharmacological blockade endocytosis. Together, our data support a model where contributes regulation secretion an endocytosis-dependent manner reprogram contents microdomains.

Язык: Английский

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Tetraspanins interweave EV secretion, endosomal network dynamics and cellular metabolism

European Journal of Cell Biology, Год журнала: 2022, Номер 101(3), С. 151229 - 151229

Опубликована: Апрель 27, 2022

Tetraspanin proteins organize membrane nanodomains related to cell adhesion and migration. An essential feature conserved along the superfamily is their cone-shaped tertiary structure, which allows tetraspanins be enriched in highly curved structures. Their conical shape, together with ability associate transmembrane receptors bind cystoskeletal signaling scaffolds, are key regulate endosomal network dynamics Extracellular Vesicle biogenesis cargo selection. Recent evidence suggests that have a relevant impact mitochondria turnover regulation of cellular metabolism. In this review we highlight those reports point as regulators communication between network, EVs

Язык: Английский

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Prognostic value and multifaceted roles of tetraspanin CD9 in cancer

Frontiers in Oncology, Год журнала: 2023, Номер 13

Опубликована: Март 17, 2023

CD9 is a crucial regulator of cell adhesion in the immune system and plays important physiological roles hematopoiesis, blood coagulation or viral bacterial infections. It involved transendothelial migration leukocytes which might also be hijacked by cancer cells during their invasion metastasis. found at surface membrane exosomes affecting progression therapy resistance. High expression mostly associated with good patients outcome, few exceptions. Discordant findings have been reported for breast, ovarian, melanoma, pancreatic esophageal cancer, related to using different antibodies inherent heterogeneity. According vitro vivo studies, tetraspanin not clearly either tumor suppression promotion. Further mechanistic experiments will elucidate role particular types specific conditions.

Язык: Английский

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CD9 inhibition reveals a functional connection of extracellular vesicle secretion with mitophagy in melanoma cells

Journal of Extracellular Vesicles, Год журнала: 2021, Номер 10(7)

Опубликована: Май 1, 2021

Abstract Tetraspanins are often used as Extracellular Vesicle (EV) detection markers because of their abundance on these secreted vesicles. However, data function EV biogenesis controversial and compensatory mechanisms occur upon gene deletion. To overcome this handicap, we have compared the effects tetraspanin CD9 deletion with those elicited by cytopermeable peptides blocking properties against CD9. Both peptide or reduced number early endosomes. induced an increase in lysosome numbers, while augmented MVB secretion, probably CD63 expression upregulation. In vivo , delayed primary tumour cell growth metastasis size. These proliferation were shown to be concomitant impairment mitochondrial quality control. KO cells able compensate malfunction increasing total mass reducing mitophagy. Our thus provide first evidence for a functional connection mitophagy melanoma cells.

Язык: Английский

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Asymmetric depth‐filtration: A versatile and scalable method for high‐yield isolation of extracellular vesicles with low contamination

Journal of Extracellular Vesicles, Год журнала: 2022, Номер 11(8)

Опубликована: Авг. 1, 2022

Abstract We developed a novel asymmetric depth filtration (DF) approach to isolate extracellular vesicles (EVs) from biological fluids that outperforms ultracentrifugation and size‐exclusion chromatography in purity yield of isolated EVs. By these metrics, single‐step DF matches or exceeds the performance multistep protocols with dedicated purification procedures isolation plasma demonstrate selective transit capture nanoparticles pores by size elasticity, low surface binding medium, ability cleanse EVs held filter before their recovery reversed flow all contribute achieved preparations. further method's versatility applying it different biofluids (plasma, urine, cell culture growth medium). The workflow is simple, fast, inexpensive. Only standard laboratory equipment required for its implementation, making suitable low‐resource point‐of‐use locations. method may be used EV small samples diagnostic treatment guidance applications. It can also scaled up harvest therapeutic large volumes medium.

Язык: Английский

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Extracellular vesicles in the glioblastoma microenvironment: A diagnostic and therapeutic perspective

Molecular Aspects of Medicine, Год журнала: 2022, Номер 91, С. 101167 - 101167

Опубликована: Дек. 27, 2022

Язык: Английский

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ApoE expression in macrophages communicates immunometabolic signaling that controls hyperlipidemia‐driven hematopoiesis & inflammation via extracellular vesicles

Journal of Extracellular Vesicles, Год журнала: 2023, Номер 12(8)

Опубликована: Авг. 1, 2023

Abstract While apolipoprotein E (apoE) expression by myeloid cells is recognized to control inflammation, whether such benefits can be communicated via extracellular vesicles not known. Through the study of produced macrophages derived from bone marrow Wildtype (WT‐BMDM‐EV) and ApoE deficient (EKO‐BMDM‐EV) mice, we uncovered a critical role for apoE in regulating their cell signaling properties. WT‐BMDM‐EV anti‐inflammatory properties recipient increasing cellular levels miR‐146a‐5p, that reduced NF‐κB signalling. They also downregulated miR‐142a‐3p, resulting increased its target carnitine palmitoyl transferase 1A (CPT1A) which improved fatty acid oxidation (FAO) oxidative phosphorylation (OxPHOS) cells. Such favorable metabolic polarization enhanced cell‐surface MerTK phagocytic uptake apoptotic In contrast, EKO‐BMDM‐EV exerted opposite effects reducing NF‐κB−driven GLUT1‐mediated glucose uptake, aerobic glycolysis, stress. Furthermore, miR‐142a‐3p levels, CPT1A impaired FAO OxPHOS When cultured with naïve CD4 + T lymphocytes, drove activation proliferation, fostered transition Th1 phenotype. infusions into hyperlipidemic mice resolved hematopoiesis inflammatory responses lymphocytes. ApoE‐dependent immunometabolic macrophage was dependent on transcriptional axes controlled miR‐146a‐5p could reproduced infusing miR‐146a mimics & miR‐142a antagonists apoE‐deficient mice. Together, our findings unveil novel property modulates regulatory secreted vesicles.

Язык: Английский

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