Failure to detect functional transfer of active K-Ras protein from extracellular vesicles into recipient cells in culture

Luhtala, Natalie; Hunter, Tony

doi:10.1371/journal.pone.0203290

Cited by 7 publications

(6 citation statements)

References 79 publications

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“… [ 155 ] CLIC1 Up GFP, FLAG-tagged CLIC1 is a circulating protein, secreted via extracellular vehicles (Evs) released by either cell lines or GBM-derived CSCs. [ 156 ] K-Ras Up Raf-RBD – [ 157 ] immunoglobulin (Ig) G2 and IgG4 Up CD163 – [ 158 ] TrkB Up YKL-40 Inhibits tumor growth in vivo. Plays a key role in the control of GBM progression and aggressiveness.…”

Section: Exosomal Micrornas and Gliomasmentioning

confidence: 99%

“…Tumor-derived proteins that are freely circulating in blood may be highly diluted, and could be mixed with other similar circulating biomolecules, which could confuse the tests [ 177 , 178 ]. It was found that many exosomal proteins such as HMGB 1[ 140 ], IL-8, PDGFs, caveolin 1, and lysyl oxidase [ 141 ], L1CAM [ 142 ], STC1, STC2 [ 143 ], EGFRvIII [ 144 ], VEGF-A [ 145 ], CRYAB [ 146 ], PTRF [ 147 ], PD- 1[ 148 ], IL13Rα2, IL13QD [ 149 ], CAV 1 [ 150 ], NK-Exo [ 151 ], SRSF1, SRSF3 [ 152 ], NANOGP8 [ 153 ], PTENP1 [ 155 ], CLIC1 [ 156 ], K-Ras [ 157 ], immunoglobulin (Ig) G2 and IgG4 [ 158 ], TrkB [ 159 ], MGMT mRNA [ 160 ], EGFRvIII [ 161 ], N-glycoproteins [ 162 ], LOX, ADAMTS1, TSP1, VEGF [ 163 ], NF-κB [ 165 ], Glut-1, HK-2, and PKM-2 [ 166 ], TDP-43 [ 167 ] were up-regulated. On the other hand some other proteins, such as IL-8, ZAP70, TGF-β [ 148 ], IFN-gamma, granzyme B [ 154 ], and CRCL [ 164 ] were down-regulated in glioma tumors (Table 2 ).…”

Section: Exosomal Proteins In Gliomamentioning

confidence: 99%

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Role of exosomes in malignant glioma: microRNAs and proteins in pathogenesis and diagnosis

Mahjoubin‐Tehran

Movahedpour

et al. 2020

Cell Commun Signal

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Malignant gliomas are the most common and deadly type of central nervous system tumors. Despite some advances in treatment, the mean survival time remains only about 1.25 years. Even after surgery, radiotherapy and chemotherapy, gliomas still have a poor prognosis. Exosomes are the most common type of extracellular vesicles with a size range of 30 to 100 nm, and can act as carriers of proteins, RNAs, and other bioactive molecules. Exosomes play a key role in tumorigenesis and resistance to chemotherapy or radiation. Recent evidence has shown that exosomal microRNAs (miRNAs) can be detected in the extracellular microenvironment, and can also be transferred from cell to cell via exosome secretion and uptake. Therefore, many recent studies have focused on exosomal miRNAs as important cellular regulators in various physiological and pathological conditions. A variety of exosomal miRNAs have been implicated in the initiation and progression of gliomas, by activating and/or inhibiting different signaling pathways. Exosomal miRNAs could be used as therapeutic agents to modulate different biological processes in gliomas. Exosomal miRNAs derived from mesenchymal stem cells could also be used for glioma treatment. The present review summarizes the exosomal miRNAs that have been implicated in the pathogenesis, diagnosis and treatment of gliomas. Moreover, exosomal proteins could also be involved in glioma pathogenesis. Exosomal miRNAs and proteins could also serve as non-invasive biomarkers for prognosis and disease monitoring.

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Section: Exosomal Micrornas and Gliomasmentioning

confidence: 99%

Section: Exosomal Proteins In Gliomamentioning

confidence: 99%

Role of exosomes in malignant glioma: microRNAs and proteins in pathogenesis and diagnosis

Mahjoubin‐Tehran

Movahedpour

et al. 2020

Cell Commun Signal

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“…[ 15,18 ] Although CRE recombinase has an efficiency of 55%, [ 19 ] horizontal transfer was shown to happen mainly between tumor cells in vivo, but not between tumor cells and host cells. [ 15,18 ] These results have been further confirmed using other genetic strategies, [ 20,21 ] suggesting that surface binding may represent the main mechanism of EV‐mediated tumor–host communications in vivo.…”

Section: Introductionmentioning

confidence: 71%

Cell Surface Labeling by Engineered Extracellular Vesicles

et al. 2020

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Deoxycytidine [3]). Extracellular vesicles (EVs) are increasingly recognized as a new class of multivalent messengers involved in cell-cell communication, both locally and at distance. [4-7] EVs are membranebound particles released by virtually every cell type, with stressed and transformed cells secreting the highest amounts. [8-11] Investigations of EVs' roles in different pathophysiological conditions are thwarted by biases introduced during isolation and reinfusion procedures. In particular, these biases manifest at different levels: (i) certain EV subsets may be excluded, [6] (ii) operator judgment is involved in deciding how many EVs to reinfuse, and (iii) an assumption that EV concentration is highest in blood, whereas several studies have shown that EV biodistribution occurs primarily via lymph fluid. [12-15] To circumvent these issues, the EV-releasing cells of interest can be genetically engineered to express membrane-bound reporters (or other functional biomolecules) in vitro or in vivo. [16] Although this approach is still in its infancy, it avoids the above limitations and to unbiasedly investigate untouched, endogenously released EVs. Although a detailed description of how EVs signal between cells is still missing, two nonexclusive mechanisms have been proposed: (i) membrane fusion between EVs and target cells, thereby releasing EV cargo (e.g., microRNAs and proteins, including oncogenes) directly in the cytoplasm of targeted cells; and (ii) surface binding with subsequent ligand-receptor type signaling. Membrane fusion (also known as horizontal transfer) has been described using several in vitro models (see Pucci and Pittet [17] and references therein), but proved more challenging to demonstrate in vivo. Recently, horizontal transfer of tumor-derived EVs was tested in mice using a sensitive and stringent approach based on the CRE-Lox reporter system. [15,18] Although CRE recombinase has an efficiency of 55%, [19] horizontal transfer was shown to happen mainly between tumor cells in vivo, but not between tumor cells and host cells. [15,18] These results have been further confirmed using other genetic strategies, [20,21] suggesting that surface binding may represent the main mechanism of EV-mediated tumor-host communications in vivo. A major limitation in the study of native EV signaling to host cells in vivo derives from their small size, which restricts the amount of reporter molecules carried by a single EV. As a consequence, only target cells binding the highest levels of EVs [12,14,15] will accumulate enough reporter molecules to be detected and investigated. Nonetheless, many host cell types Extracellular vesicles (EVs) can mediate local and long-range intercellular communication via cell surface signaling. In order to perform in vivo studies of unmanipulated, endogenously released EVs, sensitive but stringent approaches able to detect EV-cell surface interactions are needed. However, isolation and reinfusion of EVs can introduce biases. A rigorous way to study EVs in vivo is by genetically eng...

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“…We reported an increase in the TP53 copy number in LPSderived exosomes, suggesting that the mechanism behind DNA sorting is not stochastic. Of note, it has recently been shown that functional KRAS protein transfer from cell to cell is an extremely rare event [29]. To date, the mechanisms by which nucleic acids are sorted and packaged within EVs have yet to be elucidated [30].…”

Section: Discussionmentioning

confidence: 99%

Pro-Inflammatory Microenvironment Modulates the Transfer of Mutated TP53 Mediated by Tumor Exosomes

Domenis

Cifù

Mio

et al. 2021

IJMS

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Exosomes released from tumor cells are instrumental in shaping the local tumor microenvironment to allow cancer progression. Recently, it has been shown that tumor exosomes carry large fragments of dsDNA, which may reflect the mutational status of parental cells. Although it has been described that a stressful microenvironment can influence exosomal cargo, the effects on DNA packing and its transfer into recipient cells have yet to be investigated. Here, we report that exosomes derived from SW480 (human colorectal adenocarcinoma cell line) cells can carry dsDNA fragments containing the entire coding sequence of both TP53 and KRAS genes, harboring the SW480-related TP53 c.818G > A and KRAS c.35G > T typical mutations. We also report the following: that cell stimulation with lipopolysaccharides (LPS) promotes the selective packaging of the TP53 gene, but not the KRAS gene; that exosomes secreted by SW480 cells efficiently transfer the mutated sequences into normal CCD841-CoN colon epithelial and THLE-2 hepatic cells; that this mechanism is more efficient when the cells had been previously incubated with pro-inflammatory cytokines; that the TP53 gene appears actively transcribed in both recipient cells; and that mutated mRNA levels are not influenced by cytokine treatment. Our data strongly suggest that pro-inflammatory stimulation promotes the horizontal transfer of an oncogene by exosomes, although this remains a rare event. Further studies are needed to assess the impact of the oncogenic transfer by exosomes in malignant transformation and its role in tumor progression.

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Failure to detect functional transfer of active K-Ras protein from extracellular vesicles into recipient cells in culture

Cited by 7 publications

References 79 publications

Role of exosomes in malignant glioma: microRNAs and proteins in pathogenesis and diagnosis

Role of exosomes in malignant glioma: microRNAs and proteins in pathogenesis and diagnosis

Cell Surface Labeling by Engineered Extracellular Vesicles

Pro-Inflammatory Microenvironment Modulates the Transfer of Mutated TP53 Mediated by Tumor Exosomes

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