Journal: International Journal of Molecular Sciences

Article Title: Chloramidine/Bisindolylmaleimide-I-Mediated Inhibition of Exosome and Microvesicle Release and Enhanced Efficacy of Cancer Chemotherapy

doi: 10.3390/ijms18051007

Figure Lengend Snippet: Pharmacological inhibition of EMV release from PC3 cells is highest with Cl-amidine, bisindolylmaleimide-I, and imipramine. ( A ) Using NTA, the most significant inhibition of EMV release from PC3 cells after 24 h was observed in the presence of Cl-amidine, bisindolylmaleimide-I, and imipramine. After 24 h, none of the inhibitors caused any significant reduction in cell viability, ( B ) except for d -pantethine. The experiments were repeated three times, and the data presented are mean ± SEM of the results. (* p ≤ 0.05; **** p ≤ 0.0001).

Article Snippet: The Effect of EMV Inhibitors on 5-FU-Mediated Apoptosis of PC3 and MCF-7 Cells Combinations of EMV inhibitors bisindolylmaleimide-I and Cl-amidine were used to test for a synergistic enhancement of the anti-cancer drug, 5-fluorouracil (5-FU; Sigma-Aldrich).

Techniques: Inhibition

Journal: International Journal of Molecular Sciences

Article Title: Chloramidine/Bisindolylmaleimide-I-Mediated Inhibition of Exosome and Microvesicle Release and Enhanced Efficacy of Cancer Chemotherapy

doi: 10.3390/ijms18051007

Figure Lengend Snippet: Cl-amidine and bisindolylmaleimide-1-mediated inhibition of EMV release increases the apoptosis of PC3 and MCF-7 cells treated with 5-FU. The Guava Viacount Cell Death Assay shows that PC3 and MCF-7 cells that were given 5-FU together with Cl-amidine, bisindolylmaleimide-I, or with a combination of Cl-amidine and bisindolylmaleimide-I, had significantly reduced levels of cell viability within 24 h compared to PC3 and MCF-7 cells receiving no EMV inhibitors and given only 5-FU. Bisindolylmaleimide-I and Cl-amidine had no significant effect on cell viability on their own. Data presented are the mean ± SEM of three independent experiments performed in triplicate (** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001 were considered statistically significant compared to the drug-treated control in the absence of inhibitors).

Article Snippet: The Effect of EMV Inhibitors on 5-FU-Mediated Apoptosis of PC3 and MCF-7 Cells Combinations of EMV inhibitors bisindolylmaleimide-I and Cl-amidine were used to test for a synergistic enhancement of the anti-cancer drug, 5-fluorouracil (5-FU; Sigma-Aldrich).

Techniques: Inhibition

Journal: International Journal of Molecular Sciences

Article Title: Chloramidine/Bisindolylmaleimide-I-Mediated Inhibition of Exosome and Microvesicle Release and Enhanced Efficacy of Cancer Chemotherapy

doi: 10.3390/ijms18051007

Figure Lengend Snippet: Nanoparticle tracking analysis (NTA) of exosomes and microvesicles (EMVs) released from PC3 cells in the presence of a range of EMV inhibitors. Plots presenting NTA analysis show the concentration of vesicles (0–900 nm in diameter) released from PC3 cells in the absence of any EMV inhibitors ( A ); In ( B ) the EMVs are shown to comprise exosomes and microvesicles (MVs) by electron microscopy, by Western blotting (for CD63 expression), and by the degree of phosphatidylserine (PS) exposition. NTA analysis for released EMVs from PC3 cells are presented in the presence of Cl-amidine ( C ); bisindolylmaleimide-I ( D ); and imipramine ( E ). Vesicles outside the size range of 0–900 nm were excluded to avoid including larger vesicles such as MV aggregates or apoptotic bodies. The experiment was repeated three times in total (error bars ± SEM, indicated in red).

Article Snippet: The Effect of EMV Inhibitors on 5-FU-Mediated Apoptosis of PC3 and MCF-7 Cells Combinations of EMV inhibitors bisindolylmaleimide-I and Cl-amidine were used to test for a synergistic enhancement of the anti-cancer drug, 5-fluorouracil (5-FU; Sigma-Aldrich).

Techniques: Concentration Assay, Electron Microscopy, Western Blot, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Vesicular Transport of Encapsulated microRNA between Glial and Neuronal Cells

doi: 10.3390/ijms21145078

Figure Lengend Snippet: Highly schematized representation of ( A ) the release of exosomes (EXs) and ( B ) the release and uptake of extracellular microvesicles (EMVs) in neural cells of the human central nervous system (CNS); both types of vesicular transport systems have been observed to operate in the brain between astroglial cells and neurons; ( A ) exosomes—when mature intracellular endosomes (also known as multi-vesicular bodies) containing intraluminal vesicles (ILVs; black outlined green spheres) fuse with the plasma membrane and empty their plasma membrane-encapsulated cargo, ILVs are released and, from being extracellular, they become exosomes (EX); these 30–100 nm diameter spheres contain various mixtures of proteins, lipids, proteolipids, cytokines, chemokines, microRNAs (miRNA), messenger RNAs (mRNA) and end-stage neurotoxic metabolic products, including 42 amino acid amyloid-beta (Aβ42) peptides, tau proteins and/or the lipid raft associated flotillin (Angelopoulou et al. 2020 [ 29 ]; Hornung et al., 2020 [ 30 ]); EXs appear to play a central role in the spread of Aβ42 pathology and amyloidogenesis (Mathews and Levy 2019 [ 15 ]; Arbo et al., 2020 [ 6 ]; Peng et al., 2020 [ 31 ]); ( B ) extracellular microvesicles (EMVs)—the exterior plasma membrane of activated microglia (AM) can release (R) 100–1000 nm diameter EMVs directly from the outward blebbing of the plasma membrane of microglias and astrocytes and carry intracellular contents from their cells of origin; this includes various complex mixtures of proteins, lipids, proteolipids, miRNAs, mRNAs, end-stage metabolic products and cytokines and chemokines; together these EMV contents may be pathogenic and cause the spread of pro-inflammatory signaling and inflammatory neurodegeneration (Prada et al., 2018 [ 21 ]; Serpente et al., 2020 [ 22 ]; Vanherle et al., 2020 [ 4 ]). Microvesicular trafficking and the extracellular microvesicle uptake (U) by neurons (N) via directed translocation mechanisms (dashed black lines with black arrowheads) may occur via the direct fusion of the EMV membrane with the neuronal cell plasma membrane or by endocytosis (Stahl et al., 2019 [ 5 ]; Arbo et al., 2020 [ 6 ]; Hornung et al., 2020 [ 30 ]; Peng et al., 2020 [ 32 ]; Song et al., 2020 [ 9 ]; Upadhya et al., 2020 [ 20 ]).

Article Snippet: There are practical challenges associated with the methodology of the extraction and characterization of CNS-derived blood, CSF and tissue EXs and EMVs, and subsequent analysis of their miRNAs and other intra-vesicular cargoes, however the methodologies for EX and EMV isolation and categorization are constantly improving (Hornung et al., 2020 [ ]; Serpente et al., 2020 [ ]).

Techniques: Translocation Assay

Journal: Frontiers in Pharmacology

Article Title: Cannabidiol (CBD) Is a Novel Inhibitor for Exosome and Microvesicle (EMV) Release in Cancer

doi: 10.3389/fphar.2018.00889

Figure Lengend Snippet: CBD significantly inhibits total EMV, exosome and MV release from PC3 cells. Inhibitory effects of CBD alone and in combination with Cl-amidine on extracellular vesicle release from PC3 cancer cells are presented as histograms which are based on size exclusion analysis by Nanosight Tracking Analysis (NTA). EMVs represent all vesicles 0–900 nm (A) ; exosomes are vesicles

Article Snippet: Isolation of EMVs Exosome and microvesicles were isolated from the CBD, Cl-amidine, and CBD plus Cl-amidine treated cell culture supernatants, as well as from the control treated cells (DMSO or PBS), by differential centrifugation as follows: First, whole cells were removed by spinning at 200 g /5 min at 4°C.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Cannabidiol (CBD) Is a Novel Inhibitor for Exosome and Microvesicle (EMV) Release in Cancer

doi: 10.3389/fphar.2018.00889

Figure Lengend Snippet: CBD significantly inhibits total EMV, exosome and MV release from HEPG2 cells. Inhibitory effects of CBD alone and in combination with Cl-amidine on extracellular vesicle release from HEPG2 cancer cells are presented as histograms which are based on size exclusion analysis by Nanosight Tracking Analysis (NTA). EMVs represent all vesicles 0–900 nm (A) ; exosomes are vesicles

Article Snippet: Isolation of EMVs Exosome and microvesicles were isolated from the CBD, Cl-amidine, and CBD plus Cl-amidine treated cell culture supernatants, as well as from the control treated cells (DMSO or PBS), by differential centrifugation as follows: First, whole cells were removed by spinning at 200 g /5 min at 4°C.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Cannabidiol (CBD) Is a Novel Inhibitor for Exosome and Microvesicle (EMV) Release in Cancer

doi: 10.3389/fphar.2018.00889

Figure Lengend Snippet: CBD significantly inhibits total EMV, exosome and MV release from MDA-MB-231 cells. Inhibitory effects of CBD alone and in combination with Cl-amidine on extracellular vesicle release from MDA-MB-231 cancer cells are presented as histograms which are based on size exclusion analysis by Nanosight Tracking Analysis (NTA). EMVs represent all vesicles 0–900 nm (A) ; exosomes are vesicles

Article Snippet: Isolation of EMVs Exosome and microvesicles were isolated from the CBD, Cl-amidine, and CBD plus Cl-amidine treated cell culture supernatants, as well as from the control treated cells (DMSO or PBS), by differential centrifugation as follows: First, whole cells were removed by spinning at 200 g /5 min at 4°C.

Techniques: Multiple Displacement Amplification