Dataset Information

Goat’s milk exosomes as natural nanoparticles in detection of inflammatory processes by optical imaging

ABSTRACT: Exosomes are cell-derived nanovesicles with a proven key role in intercellular signaling, inflammation processes and immune response. Due to their natural origin and liposome-like structure, these nanometric vesicles have emerged as novel platforms for therapy and diagnosis. This work presents goat’s milk exosomes as natural nanoparticles applied in the detection of inflammatory processes by optical imaging. Goat’s milk exosomes were isolated and fully characterized by their physicochemical properties, proteomics profile and biochemical impact in healthy mice. For the in vitro and in vivo assessment, these nanoparticles were covalently labeled with sulfo-Cyanine 5 and BODIPY-FL. In vitro studies confirmed the internalization of the nanoprobe as well as no cytotoxicity in macrophages populations (RAW 264.7). The capacity of our probe to localize inflammatory processes was in vivo assessed by optical imaging in a peritonitis mouse model confirmed. These results were also confirmed by the flow cytometry analysis of the peritonitis exudates, supporting the use of these natural nanoparticles in the detection of inflammatory processes.

INSTRUMENT(S): Q Exactive Plus

ORGANISM(S): Capra Hircus (goat)

TISSUE(S): Milk

DISEASE(S): Inflammation

SUBMITTER: Pilar Ximenez-Embun

LAB HEAD: Javier Muñoz Peralta

PROVIDER: PXD025026 | Pride | 2022-02-21

REPOSITORIES: Pride

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Publications

Goat Milk Exosomes As Natural Nanoparticles for Detecting Inflammatory Processes By Optical Imaging.

Santos-Coquillat Ana A González María Isabel MI Clemente-Moragón Agustín A González-Arjona Mario M Albaladejo-García Virginia V Peinado Héctor H Muñoz Javier J Ximénez Embún Pilar P Ibañez Borja B Oliver Eduardo E Desco Manuel M Salinas Beatriz B

Small (Weinheim an der Bergstrasse, Germany) 20211202 6

Exosomes are cell-derived nanovesicles with a proven intercellular signaling role in inflammation processes and immune response. Due to their natural origin and liposome-like structure, these nanometer-scale vesicles have emerged as novel platforms for therapy and diagnosis. In this work, goat milk exosomes are isolated and fully characterized in terms of their physicochemical properties, proteomics, and biochemical profile in healthy mice, and used to detect inflammatory processes by optical im ...[more]

PMID: 34854563

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Project description:Background: Exosomes are nanovesicles of endocytic origin believed to be involved in communication between cells. Recently, it has been shown that mast cell exosomes contain RNA named "exosomal shuttle RNA". The aim of this study was to evaluate whether exosomal shuttle RNA could play a role in the communication between human mast cells and between human mast cells and human CD34 positive progenitor cells. Results: Exosomes from the human mast cell line HMC-1 contain RNA. The exosomes contain no or very little ribosomal RNA compared to their donor cells. The mRNA and microRNA content in exosomes and their donor cells was examined using microarray analyses. We found 116 microRNA in the exosomes and 134 microRNA in the cells, from which some were expressed at different level. DNA microarray experiments revealed the presence of approximately 1800 mRNAs in the exosomes, which represent 15% of the donor cell mRNA content. Transfer experiments revealed that exosomes and their RNA can transfer to other HMC-1 cells and to CD34 positive progenitors. Conclusions: To conclude, HMC-1 exosomes contain mRNA and microRNA that can be transferred to other mast cells and to CD34 progenitors. This shuttle of exosomal RNA may represent a powerful mode of communication between cells where cells send genetic information to other cells over a distance via exosomes. [miRNA profiling] Identification of microRNA was performed by Exiqon (www.exiqon.com). Briefly, the quality of the total RNA was verified by an Agilent 2100 Bioanalyzer. Total RNA from the exosome and the HMC-1 cell samples were labelled with Hy3 and Hy5 fluorescent stain, respectively, using the miRCURY Hy3/Hy5 power labelling kit. The Hy3-labelled exosome samples and a Hy5-labeled mast cells were mixed pair-wise and hybridized to the miRCURY? LNA array (v9.2). The hybridization was performed according to the miRCURY? LNA array manual using a Tecan HS4800 hybridization station (Tecan Systems, Inc. San Jose, CA). The miRCURY? LNA array microarray slides were scanned by a ScanArray 4000 XL scanner (Packard Biochip Technologies, Billerica, MA ,USA) and the image analysis was carried out using the ImaGene 6.1.0 software (BioDiscovery, Inc, El Segundo, CA USA). The quantified signals were normalized using the global Lowess (LOcally WEighted Scatterplot Smoothing) regression algorithm. MicroRNA with signals equal to or below the background signal in 2 or more of the 4 replicate measurements were identified as absent in that slide. The limit for a miRNA to be listed as detectable was set to signal intensities higher than 3 x background (3 x median Hy3 or Hy5 for the total slide). In addition, where signals were detected for <3 of the slides, they were considered unreliable and excluded from sets of detected miRNAs. The experiment was performed in triplicate samples. The signal was calculated as the mean value of the log2MeanRatio Hy3/Hy5 for the triplicates M-BM-1 SD. [mRNA profiling] Exosomes were prepared from the supernatant of HMC-1 cells by differential centrifugations and filtration. RNA was isolated from the exosomes and their parental cells using Trizol . The microarray experiments were performed by SweGene (www.swegene.org/) according to Affymetrix microarray DNA chip analysis (n=4). p0739_E1, p0739_ E2, p0739_E3 and p0739_E4 for the exosomes samples and p0739_C1, p0739_C2, p0739_C3, and p0739_C4 for the HMC-1 cells. [miRNA profiling] Exosomes were prepared from the supernatant of HMC-1 cells by differential centrifugations and filtration. RNA was isolated from the exosomes and their parental cells using Trizol followed by RNeasy clean-up. The microarray experiments were performed by Exiqon.

Dataset Information

Goat’s milk exosomes as natural nanoparticles in detection of inflammatory processes by optical imaging

Publications

Goat Milk Exosomes As Natural Nanoparticles for Detecting Inflammatory Processes By Optical Imaging.

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