bims-exocan 2022-08-14 papers

bims-exocan

Biomed News

on Exosomes roles in cancer

Issue of 2022‒08‒14
38 papers selected by
Muhammad Rizwan
COMSATS University

Role of exosomes in lung cancer: A comprehensive insight from immunomodulation to theragnostic applications.
Roles of exosomes as drug delivery systems in cancer immunotherapy: a mini-review.
Current status and outlook of advances in exosome isolation.
Recent advances in nanotechnology-enabled biosensors for detection of exosomes as new cancer liquid biopsy.
Selective isolation of extracellular vesicles from minimally processed human plasma as a translational strategy for liquid biopsies.
Harnessing the Full Potential of Extracellular Vesicles as Drug Carriers.
Blockade of exosome generation by GW4869 inhibits the education of M2 macrophages in prostate cancer.
Extracellular vesicle-mediated immunoregulation in cancer.
A Novel Localization in Human Large Extracellular Vesicles for the EGF-CFC Founder Member CRIPTO and Its Biological and Therapeutic Implications.
Cyclooxygenase-2 Upregulated by Temozolomide in Glioblastoma Cells Is Shuttled In Extracellular Vesicles Modifying Recipient Cell Phenotype.
Adipocyte-Derived Extracellular Vesicles Promote Prostate Cancer Cell Aggressiveness by Enabling Multiple Phenotypic and Metabolic Changes.
Serum Extracellular Vesicle Stratifin Is a Biomarker of Perineural Invasion in Patients With Colorectal Cancer and Predicts Worse Prognosis.
Extracellular Vesicles in Myeloid Neoplasms.
Tumor-derived extracellular vesicles for the active targeting and effective treatment of colorectal tumors in vivo.
Exosomal microRNA-551b-3p from bone marrow-derived mesenchymal stromal cells inhibits breast cancer progression via regulating TRIM31/Akt signaling.
Overcoming the blood-brain barrier: Exosomes as theranostic nanocarriers for precision neuroimaging.
Asymmetric depth-filtration: A versatile and scalable method for high-yield isolation of extracellular vesicles with low contamination.
Metastasis suppressor NME1 in exosomes or liposomes conveys motility and migration inhibition in breast cancer model systems.
Exosome Metabolic Patterns on Aptamer-Coupled Polymorphic Carbon for Precise Detection of Early Gastric Cancer.
Thymoquinone-loaded mesenchymal stem cell-derived exosome as an efficient nano-system against breast cancer cells.
Exosomes from EGFR-Mutated Adenocarcinoma Induce a Hybrid EMT and MMP9-Dependant Tumor Invasion.
Human Milk Extracellular Vesicles: A Biological System with Clinical Implications.
Find new channel for overcoming chemoresistance in cancers: Role of stem cells-derived exosomal microRNAs.
A Liquid Biopsy in Bladder Cancer-The Current Landscape in Urinary Biomarkers.
Protein Panel of Serum-Derived Small Extracellular Vesicles for the Screening and Diagnosis of Epithelial Ovarian Cancer.
Detailed Characterization of Small Extracellular Vesicles from Different Cell Types Based on Tetraspanin Composition by ExoView R100 Platform.
Biosynthetic Dendritic Cell-Exocytosed Aggregation-Induced Emission Nanoparticles for Synergistic Photodynamic Immunotherapy.
Selective Internal Radiotherapy Changes the Immune Profiles of Extracellular Vesicles and Their Immune Origin in Patients with Inoperable Cholangiocarcinoma.
Diagnostic value of exosomes in patients with liver cancer: a systematic review.
Microfluidic amperometric immunosensor based on porous nanomaterial towards claudin7 determination for colorectal cancer diagnosis.
Hepatic stellate cell exosome-derived circWDR25 promotes the progression of hepatocellular carcinoma via the miRNA-4474-3P-ALOX-15 and EMT axes.
Spotlight on Exosomal Non-Coding RNAs in Breast Cancer: An In Silico Analysis to Identify Potential lncRNA/circRNA-miRNA-Target Axis.
A cryostat-based frozen section method to increase the yield of extracellular vesicles extracted from different tissues.
Identification of exosomal circRNA CD226 as a potent driver of nonsmall cell lung cancer through miR-1224-3p/high mobility group AT-hook 2 axis.
Human platelet lysate-derived extracellular vesicles enhance angiogenesis through miR-126.
Therapeutic exosomes loaded with SERPINA5 attenuated endometrial cancer cell migration via the integrin β1/FAK signaling pathway.
The Hepatic Pre-Metastatic Niche.
A systematic review of EV-piRNA in human body fluid and its role in disease progression.

Biochim Biophys Acta Rev Cancer. 2022 Aug 09. pii: S0304-419X(22)00101-9. [Epub ahead of print] 188776

Role of exosomes in lung cancer: A comprehensive insight from immunomodulation to theragnostic applications.

Faizan Haider Khan, Malik Johid Reza, Yusra Fatima Shao, Ahmad Perwez, Honey Zahra, Afshin Dowlati, Ata Abbas.

  Exosomes are 30 to 150 nm-diameter lipid bilayer-enclosed extracellular vesicles that enable cell-to-cell communication through secretion and uptake. The exosomal cargoes contain RNA, lipids, proteins, and metabolites which can be delivered to recipient cells in vivo. In a healthy lung, exosomes facilitate interaction between adaptive and innate immunity and help maintain normal lung physiology. However, tumor-derived exosomes in lung cancer (LC) can, on the other hand, restrict immune cell proliferation, cause apoptosis in activated CD8+ T effector cells, reduce natural killer cell activity, obstruct monocyte differentiation, and promote proliferation of myeloid-derived suppressor and regulatory T cells. In addition, exosomes in the tumor microenvironment may also play a critical role in cancer progression and the development of drug resistance. In this review, we aim to comprehensively examine the current updates on the role of exosomes in lung carcinogenesis and their potential application as a diagnostic, prognostic, and therapeutic tool in lung cancer.

Keywords:  Exosomes; Immune cells; Lung cancer; Tumor microenvironment; Tumor-derived exosomes

DOI:  https://doi.org/10.1016/j.bbcan.2022.188776
Discov Oncol. 2022 Aug 13. 13(1): 74

Roles of exosomes as drug delivery systems in cancer immunotherapy: a mini-review.

Zhen Fang, Yixuan Ding, Zhigang Xue, Peijuan Li, Jia Li, Fei Li.

  Exosomes can be released by a variety of cells and participate in intercellular communication in many physiological processes in the body. They can be used as carriers of cancer therapeutic drugs and have natural delivery capabilities. Some biologically active substances on exosomes, such as major histocompatibility complex (MHC), have been shown to be involved in exosome-mediated anticancer immune responses and have important regulatory effects on the immune system. Exosome-based drug delivery systems hold great promise in future cancer immunotherapy. However, there are still substantial challenges to be overcome in the clinical application of exosomes as drug carriers. This article reviews the biological characteristics of exosome drug delivery systems and their potential applications and challenges in cancer immunotherapy.

Keywords:  Cancer; Drug delivery systems; Exosomes; Immunotherapy

DOI:  https://doi.org/10.1007/s12672-022-00539-5
Anal Bioanal Chem. 2022 Aug 13.

Current status and outlook of advances in exosome isolation.

Wen-Zhao Liu, Zhan-Jun Ma, Xue-Wen Kang.

  Exosomes are extracellular vesicles with a diameter ranging from 30 to 150 nm, which are an important medium for intercellular communication and are closely related to the progression of certain diseases. Therefore, exosomes are considered promising biomarkers for the diagnosis of specific diseases, and thereby, treatments based on exosomes are being widely examined. For exosome-related research, a rapid, simple, high-purity, and recovery isolation method is the primary prerequisite for exosomal large-scale application in medical practice. Although there are no standardized methods for exosome separation and analysis, various techniques have been established to explore their biochemical and physicochemical properties. In this review, we analyzed the progress in exosomal isolation strategies and proposed our views on the development prospects of various exosomal isolation techniques.

Keywords:  Exosome; Extracellular vesicle; Isolation; Microfluidic; Separation

DOI:  https://doi.org/10.1007/s00216-022-04253-7
Exp Biol Med (Maywood). 2022 Aug 06. 15353702221110813

Recent advances in nanotechnology-enabled biosensors for detection of exosomes as new cancer liquid biopsy.

Chang-Chieh Hsu, Yun Wu.

  Cancer liquid biopsy detects circulating biomarkers in body fluids, provides information that complements medical imaging and tissue biopsy, allows sequential monitoring of cancer development, and, therefore, has shown great promise in cancer screening, diagnosis, and prognosis. Exosomes (also known as small extracellular vesicles) are cell-secreted, nanosized vesicles that transport biomolecules such as proteins and RNAs for intercellular communication. Exosomes are actively involved in cancer development and progression and have become promising circulating biomarkers for cancer liquid biopsy. Conventional exosome characterization methods such as quantitative reverse transcription polymerase chain reaction (qRT-PCR) and enzyme-linked immunosorbent assay (ELISA) are limited by low sensitivity, tedious process, large sample volume, and high cost. To overcome these challenges, new biosensors have been developed to offer sensitive, simple, fast, high throughput, low sample consumption, and cost-effective detection of exosomal biomarkers. In this review, we summarized recent advances in nanotechnology-enabled biosensors that detect exosomal RNAs (both microRNAs and mRNAs) and proteins for cancer screening, diagnosis, and prognosis. The biosensors were grouped based on their sensing mechanisms, including fluorescence-based biosensors, colorimetric biosensors, electrical/electrochemical biosensors, plasmonics-based biosensors, surface-enhanced Raman spectroscopy (SERS)-based biosensors, and inductively coupled plasma mass spectrometry (ICP-MS) and photothermal biosensors. The future directions for the development of exosome-based biosensors were discussed.

Keywords:  Exosomes; biosensors; cancer liquid biopsy; extracellular vesicles; mRNA; microRNA; protein

DOI:  https://doi.org/10.1177/15353702221110813
Biomark Res. 2022 Aug 07. 10(1): 57

Selective isolation of extracellular vesicles from minimally processed human plasma as a translational strategy for liquid biopsies.

Diogo Fortunato, Stavros Giannoukakos, Ana Giménez-Capitán, Michael Hackenberg, Miguel A Molina-Vila, Nataša Zarovni.

  BACKGROUND: Intercellular communication is mediated by extracellular vesicles (EVs), as they enclose selectively packaged biomolecules that can be horizontally transferred from donor to recipient cells. Because all cells constantly generate and recycle EVs, they provide accurate timed snapshots of individual pathophysiological status. Since blood plasma circulates through the whole body, it is often the biofluid of choice for biomarker detection in EVs. Blood collection is easy and minimally invasive, yet reproducible procedures to obtain pure EV samples from circulating biofluids are still lacking. Here, we addressed central aspects of EV immunoaffinity isolation from simple and complex matrices, such as plasma.METHODS: Cell-generated EV spike-in models were isolated and purified by size-exclusion chromatography, stained with cellular dyes and characterized by nano flow cytometry. Fluorescently-labelled spike-in EVs emerged as reliable, high-throughput and easily measurable readouts, which were employed to optimize our EV immunoprecipitation strategy and evaluate its performance. Plasma-derived EVs were captured and detected using this straightforward protocol, sequentially combining isolation and staining of specific surface markers, such as CD9 or CD41. Multiplexed digital transcript detection data was generated using the Nanostring nCounter platform and evaluated through a dedicated bioinformatics pipeline.
RESULTS: Beads with covalently-conjugated antibodies on their surface outperformed streptavidin-conjugated beads, coated with biotinylated antibodies, in EV immunoprecipitation. Fluorescent EV spike recovery evidenced that target EV subpopulations can be efficiently retrieved from plasma, and that their enrichment is dependent not only on complex matrix composition, but also on the EV surface phenotype. Finally, mRNA profiling experiments proved that distinct EV subpopulations can be captured by directly targeting different surface markers. Furthermore, EVs isolated with anti-CD61 beads enclosed mRNA expression patterns that might be associated to early-stage lung cancer, in contrast with EVs captured through CD9, CD63 or CD81. The differential clinical value carried within each distinct EV subset highlights the advantages of selective isolation.
CONCLUSIONS: This EV isolation protocol facilitated the extraction of clinically useful information from plasma. Compatible with common downstream analytics, it is a readily implementable research tool, tailored to provide a truly translational solution in routine clinical workflows, fostering the inclusion of EVs in novel liquid biopsy settings.

Keywords:  Early-stage cancer; Enrichment; Extracellular vesicle; Immunoprecipitation; Liquid biopsy; Plasma; Platelet

DOI:  https://doi.org/10.1186/s40364-022-00404-1
Curr Opin Colloid Interface Sci. 2021 Feb;pii: 101412. [Epub ahead of print]51

Harnessing the Full Potential of Extracellular Vesicles as Drug Carriers.

Natalie Jasiewicz, Corinne Drabenstott, Juliane Nguyen.

  Extracellular vesicles are natural delivery systems widely implicated in cellular communication. However, to fully utilize these vehicles as nanocarriers, we must explore various methods to modify their applicability as drug delivery vehicles. In this review, we outline and discuss techniques to engineer extracellular vehicles for enhanced loading, targeting, circulation, and tracking. We highlight cutting-edge methods to amplify extracellular vesicle secretion and production and optimize storage conditions to improve their clinical suitability. Moreover, we focus on reverse engineering as an important step in controlling their biological function. By taking a reductionist approach to characterize and understand the individual components of these carriers, we can not only elucidate complex mechanisms of action but also advance the field through the creation of synthetic drug delivery vehicles. Finally, we propose current challenges and future directions of the field.

Keywords:  Cargo Loading; Extracellular vesicles; Production; Reverse Engineering; Storage; Surface Modification; Vaccine

DOI:  https://doi.org/10.1016/j.cocis.2020.101412
BMC Immunol. 2022 08 08. 23(1): 37

Blockade of exosome generation by GW4869 inhibits the education of M2 macrophages in prostate cancer.

Yilin Peng, Min Zhao, Yinying Hu, Hongyan Guo, Yanyan Zhang, Yanqin Huang, Lin Zhao, Yong Chai, Zhigang Wang.

  BACKGROUND: Tumor-associated macrophages are considered to be a major contributor affecting the development of tumors. Recently, numerous studies have shown that tumor cells were able to educate their microenvironment by delivering a significant amount of exosomes, however, the mechanism that exosomes from PCa cells work in macrophage polarization remains obscure. Therefore, we sought to determine whether blockade of exosome generation by GW4869, an inhibitor of exosome biogenesis, would impede macrophages from differentiating into M2 cells.RESULTS: In this study, we first obtained exosomes from the supernatant media of PCa cells cultured with exosome-free serum using the Magcapture™ Exosome Isolation Kit PS, and then investigated their effects on macrophages. Our data confirmed that exosomes released by prostate cancer cells can induce macrophages to differentiate into M2 cells. Mechanistically speaking, exosomes exert their effects on macrophages through activating the AKT and STAT3 signaling pathways. Importantly, treatment with GW4869 significantly inhibited the release of exosomes from PCa cells, and further impaired M2 differentiation of macrophages and their pro-tumor activity. We also demonstrated that GW4869 was able to inhibit the education of M2 macrophages, and then inhibit the progression of prostate cancer in vivo.
CONCLUSIONS: In brief, our findings indicated that GW4869 impeded the PCa exosome-induced M2 differentiation of macrophages and the progression of prostate cancer, suggesting that GW4869 could play an important role in the treatment of prostate cancer metastasis as an inhibitor of tumor exosome secretion.

Keywords:  Exosome; GW4869; Macrophage polarization; Prostate cancer

DOI:  https://doi.org/10.1186/s12865-022-00514-3
Int J Hematol. 2022 Aug 11.

Extracellular vesicle-mediated immunoregulation in cancer.

Tomofumi Yamamoto, Yusuke Yamamoto, Takahiro Ochiya.

  Extracellular vesicles (EVs) have emerged as immunomodulatory regulators during tumor progression. These small vesicles encapsulate a variety of molecules, including DNA, RNA, and proteins. When EVs come in contact with recipient cells, the EVs transmit various physiological characteristics; for example, proteins on the surface of EVs act as ligands. Immune checkpoint blockade targeting cytotoxic T-lymphocyte-associated antigen 4 (CTLA4), programmed cell death 1 (PD-1) and programmed cell death ligand 1 (PD-L1) has shown promise in a subset of cancer patients. PD-L1 on EVs acts as a key immunomodulator. Suppression of EV secretion enhances the efficacy of immunotherapy using immune checkpoint blockade antibodies. In addition to immune checkpoint blockade therapy, chimeric antigen receptor T (CAR-T) cell therapy has also been used to successfully eliminate cancer cells. Interestingly, CAR-T-cell-derived EVs express CAR on their surface. Compared with CAR-T cells, CAR-expressing EVs do not express PD1, so their antitumor effect cannot be weakened. In this review, we describe the current understanding of EVs in cancer immunity and summarize their crucial roles in immunomodulation.

Keywords:  EV-targeted therapy; Exosome; Extracellular vesicles; Immunotherapy

DOI:  https://doi.org/10.1007/s12185-022-03436-3
Cancers (Basel). 2022 Jul 29. pii: 3700. [Epub ahead of print]14(15):

A Novel Localization in Human Large Extracellular Vesicles for the EGF-CFC Founder Member CRIPTO and Its Biological and Therapeutic Implications.

Francesca Mantile, Matic Kisovec, Giorgia Adamo, Daniele P Romancino, Matej Hočevar, Darja Božič, Apolonija Bedina Zavec, Marjetka Podobnik, Maria Patrizia Stoppelli, Annamaria Kisslinger, Antonella Bongiovanni, Veronika Kralj-Iglič, Giovanna L Liguori.

  Tumor growth and metastasis strongly rely on cell-cell communication. One of the mechanisms by which tumor cells communicate involves the release and uptake of lipid membrane encapsulated particles full of bioactive molecules, called extracellular vesicles (EVs). EV exchange between cancer cells may induce phenotype changes in the recipient cells. Our work investigated the effect of EVs released by teratocarcinoma cells on glioblastoma (GBM) cells. EVs were isolated by differential centrifugation and analyzed through Western blot, nanoparticle tracking analysis, and electron microscopy. The effect of large EVs on GBM cells was tested through cell migration, proliferation, and drug-sensitivity assays, and resulted in a specific impairment in cell migration with no effects on proliferation and drug-sensitivity. Noticeably, we found the presence of the EGF-CFC founder member CRIPTO on both small and large EVs, in the latter case implicated in the EV-mediated negative regulation of GBM cell migration. Our data let us propose a novel route and function for CRIPTO during tumorigenesis, highlighting a complex scenario regulating its effect, and paving the way to novel strategies to control cell migration, to ultimately improve the prognosis and quality of life of GBM patients.

Keywords:  CRIPTO; cancer therapy; extracellular vesicles; glioblastoma; inhibition of cell migration; microvesicles; teratocarcinoma

DOI:  https://doi.org/10.3390/cancers14153700
Front Oncol. 2022 ;12 933746

Cyclooxygenase-2 Upregulated by Temozolomide in Glioblastoma Cells Is Shuttled In Extracellular Vesicles Modifying Recipient Cell Phenotype.

Francesca Lombardi, Francesca Rosaria Augello, Serena Artone, Emira Ayroldi, Ilaria Giusti, Vincenza Dolo, Maria Grazia Cifone, Benedetta Cinque, Paola Palumbo.

  Temozolomide (TMZ) resistance is frequent in patients with glioblastoma (GBM), a tumor characterized by a marked inflammatory microenvironment. Recently, we reported that cyclooxygenase-2 (COX-2) is upregulated in TMZ-resistant GBM cells treated with high TMZ concentrations. Moreover, COX-2 activity inhibition significantly counteracted TMZ-resistance of GBM cells. Extracellular vesicles (EV) are considered crucial mediators in orchestrating GBM drug resistance by modulating the tumor microenvironment (TME) and affecting the surrounding recipient cell phenotype and behavior. This work aimed to verify whether TMZ, at low and clinically relevant doses (5-20 µM), could induce COX-2 overexpression in GBM cells (T98G and U87MG) and explore if secreted EV shuttled COX-2 to recipient cells. The effect of COX-2 inhibitors (COXIB), Celecoxib (CXB), or NS398, alone or TMZ-combined, was also investigated. Our results indicated that TMZ at clinically relevant doses upregulated COX-2 in GBM cells. COXIB treatment significantly counteracted TMZ-induced COX-2 expression, confirming the crucial role of the COX-2/PGE2 system in TMZ-resistance. The COXIB specificity was verified on U251MG, COX-2 null GBM cells. Western blotting of GBM-EV cells showed the COX-2 presence, with the same intracellular trend, increasing in EV derived from TMZ-treated cells and decreasing in those derived from COXIB+TMZ-treated cells. We then evaluated the effect of EV secreted by TMZ-treated cells on U937 and U251MG, used as recipient cells. In human macrophage cell line U937, the internalization of EV derived by TMZ-T98G cells led to a shift versus a pro-tumor M2-like phenotype. On the other hand, EV from TMZ-T98G induced a significant decrease in TMZ sensitivity in U251MG cells. Overall, our results, in confirming the crucial role played by COX-2 in TMZ-resistance, provide the first evidence of the presence and effective functional transfer of this enzyme through EV derived from GBM cells, with multiple potential consequences at the level of TME.

Keywords:  COX-2; COXIB; NS398; celecoxib; extracellular vesicles; glioblastoma; temozolomide

DOI:  https://doi.org/10.3389/fonc.2022.933746
Cells. 2022 Aug 03. pii: 2388. [Epub ahead of print]11(15):

Adipocyte-Derived Extracellular Vesicles Promote Prostate Cancer Cell Aggressiveness by Enabling Multiple Phenotypic and Metabolic Changes.

Fabrizio Fontana, Martina Anselmi, Emanuela Carollo, Patrizia Sartori, Patrizia Procacci, David Carter, Patrizia Limonta.

  BACKGROUND: In recent decades, obesity has widely emerged as an important risk factor for prostate cancer (PCa). Adipose tissue and PCa cells have been shown to orchestrate a complex interaction network to support tumor growth and evolution; nonetheless, the study of this communication has only been focused on soluble factors, although increasing evidence highlights the key role of extracellular vesicles (EVs) in the modulation of tumor progression.METHODS AND RESULTS: In the present study, we found that EVs derived from 3T3-L1 adipocytes could affect PC3 and DU145 PCa cell traits, inducing increased proliferation, migration and invasion. Furthermore, conditioning of both PCa cell lines with adipocyte-released EVs resulted in lower sensitivity to docetaxel, with reduced phosphatidylserine externalization and decreased caspase 3 and PARP cleavage. In particular, these alterations were paralleled by an Akt/HIF-1α axis-related Warburg effect, characterized by enhanced glucose consumption, lactate release and ATP production.
CONCLUSIONS: Collectively, these findings demonstrate that EV-mediated crosstalk exists between adipocytes and PCa, driving tumor aggressiveness.

Keywords:  Warburg effect; adipocytes; chemoresistance; extracellular vesicles; metastasis; obesity; prostate cancer

DOI:  https://doi.org/10.3390/cells11152388
Front Oncol. 2022 ;12 912584

Serum Extracellular Vesicle Stratifin Is a Biomarker of Perineural Invasion in Patients With Colorectal Cancer and Predicts Worse Prognosis.

Wenyun Hou, Meng Pan, Yi Xiao, Wei Ge.

  Previous studies have shown that the presence of perineural invasion (PNI) is associated with a significantly worse prognosis in colorectal cancer (CRC) patients. In this study, we performed a detailed analysis of the diversity of extracellular vesicles (EV) between NPNI (non-PNI) and PNI using quantitative proteomics and aim to investigate the mechanisms underlying PNI in colorectal cancer. Quantitative proteomics technology was used to identify the proteome of serum-purified EVs from CRC patients with and without PNI (PNI and non-PNI (NPNI) groups, respectively) and healthy volunteers. Mass spectrometry data were verified by ELISA and Western blot analyses. The proteomic profile of serum EVs from the PNI group differed from that of those in the NPNI group. Serum-derived EVs from the PNI promoted more significant cellular mobility than EVs derived from the NPNI group. EV stratifin (SFN) expression levels demonstrated an area under the receiver operating characteristic curve values of 0.84 for discriminating patients with PNI from NPNI patients. Moreover, EV SFN expression levels were an independent predictor of CRC prognosis. In this study, we identified SFN as a potential biomarker for the diagnosis of PNI in stage II CRC patients.

Keywords:  colorectal cancer; epithelial-mesenchymal transition; extracellular vesicles; perineural invasion; proteomics

DOI:  https://doi.org/10.3389/fonc.2022.912584
Int J Mol Sci. 2022 Aug 08. pii: 8827. [Epub ahead of print]23(15):

Extracellular Vesicles in Myeloid Neoplasms.

Christina Karantanou, Valentina René Minciacchi, Theodoros Karantanos.

  Myeloid neoplasms arise from malignant primitive cells, which exhibit growth advantage within the bone marrow microenvironment (BMM). The interaction between these malignant cells and BMM cells is critical for the progression of these diseases. Extracellular vesicles (EVs) are lipid bound vesicles secreted into the extracellular space and involved in intercellular communication. Recent studies have described RNA and protein alterations in EVs isolated from myeloid neoplasm patients compared to healthy controls. The altered expression of various micro-RNAs is the best-described feature of EVs of these patients. Some of these micro-RNAs induce growth-related pathways such as AKT/mTOR and promote the acquisition of stem cell-like features by malignant cells. Another well-described characteristic of EVs in myeloid neoplasms is their ability to suppress healthy hematopoiesis either via direct effect on healthy CD34+ cells or via alteration of the differentiation of BMM cells. These results support a role of EVs in the pathogenesis of myeloid neoplasms. mainly through mediating the interaction between malignant and BMM cells, and warrant further study to better understand their biology. In this review, we describe the reported alterations of EV composition in myeloid neoplasms and the recent discoveries supporting their involvement in the development and progression of these diseases.

Keywords:  extracellular vesicles; myeloid malignancies

DOI:  https://doi.org/10.3390/ijms23158827
Drug Deliv. 2022 Dec;29(1): 2621-2631

Tumor-derived extracellular vesicles for the active targeting and effective treatment of colorectal tumors in vivo.

Van Du Nguyen, Ho Yong Kim, You Hee Choi, Jong-Oh Park, Eunpyo Choi.

  Colorectal cancer remains one of the main causes of cancer-related deaths worldwide. Although numerous nanomedicine formulations have been developed to tackle the disease, their low selectivity still limits effective therapeutic outcomes. In this study, we isolated extracellular vesicles (EVs) from CT26 colorectal cancer cells and 4T1 murine mammary carcinoma cells, loaded them with the chemotherapeutic agent (doxorubicin, DOX). Then we evaluated the cellular uptake of the extracellular vesicles both in 2D monolayer and 3D tumor spheroid setups using confocal laser scanning microscope and flow cytometry. In vivo tumor homing of the extracellular vesicles was verified on CT26 tumor bearing BALB/c mice using in vivo imaging system. Finally, in vivo therapeutic effects were evaluated and compared using the same animal models treated with five doses of EV formulations. CT26-EV-DOX exhibited excellent biocompatibility, a high drug-loading capacity, controlled drug release behavior, and a high capability for targeting colorectal cancer cells. In particular, we verified that CT26-EV-DOX could preferentially be up taken by their parent cells and could effectively target and penetrate 3D tumor spheroids resembling colorectal tumors in vivo in comparison with their 4T1 derived EV partner. Additionally, treatment of colorectal tumor-bearing BALB/c mice with of CT26-EV-DOX significantly inhibited the growth of the tumors during the treatment course. The developed CT26-EV-DOX nanoparticles may present a novel and effective strategy for the treatment of colorectal cancer.

Keywords:  Doxorubicin; chemotherapy; colorectal cancer; extracellular vesicle; nanoparticle drug delivery

DOI:  https://doi.org/10.1080/10717544.2022.2105444
Hum Cell. 2022 Aug 09.

Exosomal microRNA-551b-3p from bone marrow-derived mesenchymal stromal cells inhibits breast cancer progression via regulating TRIM31/Akt signaling.

Ziang Yang, Bei Xu, Sheng Wu, Weige Yang, Rongkui Luo, Shengkai Geng, Zhaochen Xin, Wen Jin, Xiong Shen, Xixi Gu, Hongwei Zhang, Hong Wang.

  Mesenchymal stromal cells (MSCs) play an important role in the development of human cancer. Meanwhile, exosomes released by MSCs can mediate cell-cell communication by delivering microRNAs (miRNAs/miRs). Hence, this study aimed to explore the role of bone marrow mesenchymal stromal cell (BMSC)-derived exosomal miR-551b-3p in breast cancer. In this study, we found that upregulation of miR-551b-5p suppressed the proliferation and migration and induced the apoptosis of breast cancer cells via downregulating tripartite motif-containing protein 31 (TRIM31). In addition, miR-551b-5p could be transferred from BMSCs to breast cancer cells via exosomes; BMSC-derived exosomal miR-551b-3p suppressed the proliferation and migration and promoted the apoptosis and oxidative stress of MDA-MB-231 cells via inhibiting TRIM31. Furthermore, a xenograft mouse model was used to explore the role of BMSC-derived exosomal miR-551b-3p in vivo. We found that BMSC-derived exosomal miR-551b-3p inhibited tumor growth in a mouse xenograft model of breast cancer in vivo. Collectively, these findings indicated that BMSC-derived exosomal miR-551b-3p could suppress the development of breast cancer via downregulating TRIM31. Thus, miR-551b-3p could serve as a potential target for the treatment of breast cancer.

Keywords:  Bone marrow mesenchymal stromal cells; Breast cancer; Exosome; Proliferation; microRNAs

DOI:  https://doi.org/10.1007/s13577-022-00753-x
J Control Release. 2022 Aug 05. pii: S0168-3659(22)00482-5. [Epub ahead of print]349 902-916

Overcoming the blood-brain barrier: Exosomes as theranostic nanocarriers for precision neuroimaging.

Chang Li, Shenghui Qin, Yu Wen, Wei Zhao, Yijie Huang, Jun Liu.

  Exosomes are cell-derived vesicles with a lipid bilayer membrane that play important roles in intercellular communication. They provide an unprecedented opportunity for the development of drug delivery nanoplatforms due to their low immunogenicity, low toxicity, biocompatibility, stability, and ability to change the functions of recipient cells. In addition, exosomes can penetrate the blood-brain barrier and then target and accumulate in relevant pathological brain regions. However, few studies have focused on the applications of exosomes as nanocarriers for use in precision neuroimaging studies. Thus, this report presents the feasibility of fabricating specific exosome-based diagnostic reagents for the application of personalized/precision radiology in the central nervous system based on important recent fundamental discoveries and technological advances.

Keywords:  Blood-brain barrier; Brain targeting; Drug delivery; Exosomes; Neurological disorders; Precision radiology

DOI:  https://doi.org/10.1016/j.jconrel.2022.08.002
J Extracell Vesicles. 2022 Aug;11(8): e12256

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

Vasiliy S Chernyshev, Roman N Chuprov-Netochin, Ekaterina Tsydenzhapova, Elena V Svirshchevskaya, Rimma A Poltavtseva, Anastasiia Merdalimova, Alexey Yashchenok, Amiran Keshelava, Konstantin Sorokin, Varlam Keshelava, Gennadiy T Sukhikh, Dmitry Gorin, Sergey Leonov, Mikhail Skliar.

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

Keywords:  asymmetric pores; depth filtration; extracellular vesicles; isolation

DOI:  https://doi.org/10.1002/jev2.12256
Clin Exp Metastasis. 2022 Aug 08.

Metastasis suppressor NME1 in exosomes or liposomes conveys motility and migration inhibition in breast cancer model systems.

Imran Khan, Brunilde Gril, Ayuko Hoshino, Howard H Yang, Maxwell P Lee, Simone Difilippantonio, David C Lyden, Patricia S Steeg.

  Tumor-derived exosomes have documented roles in accelerating the initiation and outgrowth of metastases, as well as in therapy resistance. Little information supports the converse, that exosomes or similar vesicles can suppress metastasis. We investigated the NME1 (Nm23-H1) metastasis suppressor as a candidate for metastasis suppression by extracellular vesicles. Exosomes derived from two cancer cell lines (MDA-MB-231T and MDA-MB-435), when transfected with the NME1 (Nm23-H1) metastasis suppressor, secreted exosomes with NME1 as the predominant constituent. These exosomes entered recipient tumor cells, altered their endocytic patterns in agreement with NME1 function, and suppressed in vitro tumor cell motility and migration compared to exosomes from control transfectants. Proteomic analysis of exosomes revealed multiple differentially expressed proteins that could exert biological functions. Therefore, we also prepared and investigated liposomes, empty or containing partially purified rNME1. rNME1 containing liposomes recapitulated the effects of exosomes from NME1 transfectants in vitro. In an experimental lung metastasis assay the median lung metastases per histologic section was 158 using control liposomes and 15 in the rNME1 liposome group, 90.5% lower than the control liposome group (P = 0.016). The data expand the exosome/liposome field to include metastasis suppressive functions and describe a new translational approach to prevent metastasis.

Keywords:  Breast cancer metastasis; Exosomes; Liposomes; Metastasis suppressor; NME

DOI:  https://doi.org/10.1007/s10585-022-10182-7
ACS Nano. 2022 Aug 10.

Exosome Metabolic Patterns on Aptamer-Coupled Polymorphic Carbon for Precise Detection of Early Gastric Cancer.

Haolin Chen, Chuwen Huang, Yonglei Wu, Nianrong Sun, Chunhui Deng.

  Gastric cancer (GC) presents high mortality worldwide because of delayed diagnosis. Currently, exosome-based liquid biopsy has been applied in diagnosis and monitoring of diseases including cancers, whereas disease detection based on exosomes at the metabolic level is rarely reported. Herein, the specific aptamer-coupled Au-decorated polymorphic carbon (CoMPC@Au-Apt) is constructed for the capture of urinary exosomes from early GC patients and healthy controls (HCs) and the subsequent exosome metabolic pattern profiling without extra elution process. Combining with machine learning algorithm on all exosome metabolic patterns, the early GC patients are excellently discriminated from HCs, with an accuracy of 100% for both the discovery set and blind test. Ulteriorly, three key metabolic features with clear identities are determined as a biomarker panel, obtaining a more than 90% diagnostic accuracy for early GC in the discovery set and validation set. Moreover, the change law of the key metabolic features along with GC development is revealed through making a comparison among HCs and GC at early stage and advanced stage, manifesting their monitoring ability toward GC. This work illustrates the high specificity of exosomes and the great prospective of exosome metabolic analysis in disease diagnosis and monitoring, which will promote exosome-driven precision medicine toward practical clinical application.

Keywords:  carbon; diagnosis; exosomes; gastric cancer; metabolites

DOI:  https://doi.org/10.1021/acsnano.2c05355
Iran J Basic Med Sci. 2022 Jun;25(6): 723-731

Thymoquinone-loaded mesenchymal stem cell-derived exosome as an efficient nano-system against breast cancer cells.

Mahboubeh Ebrahimian, Maryam Hashemi, Leila Etemad, Zahra Salmasi.

  Objectives: Exosomes became the subject of extensive research in drug delivery approach due to their potential applicability as therapeutic tools for cancer therapy. Thymoquinone (Tq) is an anti-cancer agent due to its great anti-proliferative effect. However, poor solubility and weak bioavailability restrict its therapeutic applications. In this study, exosomes secreted from human adipocyte-derived mesenchymal stem cells (AdMSCs) were isolated and the efficacy of a novel encapsulation method for loading of Tq was investigated. Finally, the cytotoxic effect of Tq incorporated exosomes against cancer cells was evaluated.Materials and Methods: Exosomes secreted from AdMSCs were isolated via ultracentrifugation and characterized by electron microscopy and western blotting. Then, through a novel encapsulation approach, Tq was loaded into exosomes by the combination of three methods including incubation, freeze-thawing, and surfactant treatment. Then, the encapsulation efficiency, in vitro cellular uptake, and cytotoxicity of Tq incorporated exosomes (Tq@EXOs) in MCF7 and L929 cells were estimated.
Results: Tq loading into exosomes through our novel method caused a significant improvement in encapsulation efficiency of about 60%. The fluorescent microscopy and flow cytometry outcomes indicated the efficient uptake of Tq@EXOs-FITC by cells throughout 4 hr. Furthermore, MTT results displayed the ability of Tq@EXOs in effectively decreasing the cell viability of MCF7 without causing any obvious cytotoxicity on L929 as normal cells.
Conclusion: The results suggest that our approach provides effective loading of Tq into exosomes which offer a valuable and safe platform for drug delivery to cancer cells thus having a great potential for clinical studies.

Keywords:  Adipose-derived; Breast cancer; Drug delivery system; Exosome; Thymoquinone; mesenchymal; stem cells

DOI:  https://doi.org/10.22038/IJBMS.2022.64092.14116
Cancers (Basel). 2022 Aug 03. pii: 3776. [Epub ahead of print]14(15):

Exosomes from EGFR-Mutated Adenocarcinoma Induce a Hybrid EMT and MMP9-Dependant Tumor Invasion.

Amina Jouida, Marissa O'Callaghan, Cormac Mc Carthy, Aurelie Fabre, Parthiban Nadarajan, Michael P Keane.

  Exosomes, a class of extra cellular nano-sized vesicles (EVs), and their contents have gained attention as potential sources of information on tumor detection and regulatory drivers of tumor progression and metastasis. The effect of exosomes isolated from patients with an Epidermal Growth Factor Receptor (EGFR)-mutated adenocarcinoma on the promotion of epithelial-mesenchymal transition (EMT) and invasion were examined. Exosomes derived from serum of patients with EGFR-mutated non-small cell lung cancer (NSCLC) mediate the activation of the Phosphoinositide 3-kinase (PI3K)/AKT/ mammalian target of rapamycin (mTOR) pathway and induce an invasion through the up-regulation of matrix metalloproteinase-9 (MMP-9) in A549 cells. We observed a significant increase in the expression of vimentin, a mesenchymal marker, while retaining the epithelial characteristics, as evidenced by the unaltered levels of E-cadherin and Epithelial cell adhesion molecule (EPCAM). We also observed an increase of nuclear factor erythroid 2-related factor 2 (NFR2) and P-cadherin expression, markers of hybrid EMT. Exosomes derived from EGFR-mutated adenocarcinoma serum could be a potential mediator of hybrid EMT and tumor invasion. Understanding how cancerous cells communicate and interact with their environment via exosomes will improve our understanding of lung cancer progression and metastasis formation.

Keywords:  EGFR; exosome; lung cancer; partial EMT

DOI:  https://doi.org/10.3390/cancers14153776
Cells. 2022 Jul 30. pii: 2345. [Epub ahead of print]11(15):

Human Milk Extracellular Vesicles: A Biological System with Clinical Implications.

Somchai Chutipongtanate, Ardythe L Morrow, David S Newburg.

  The consumption of human milk by a breastfeeding infant is associated with positive health outcomes, including lower risk of diarrheal disease, respiratory disease, otitis media, and in later life, less risk of chronic disease. These benefits may be mediated by antibodies, glycoproteins, glycolipids, oligosaccharides, and leukocytes. More recently, human milk extracellular vesicles (hMEVs) have been identified. HMEVs contain functional cargos, i.e., miRNAs and proteins, that may transmit information from the mother to promote infant growth and development. Maternal health conditions can influence hMEV composition. This review summarizes hMEV biogenesis and functional contents, reviews the functional evidence of hMEVs in the maternal-infant health relationship, and discusses challenges and opportunities in hMEV research.

Keywords:  breastmilk; exosomes; extracellular vesicles; human milk; maternal–child health outcomes; non-coding RNAs

DOI:  https://doi.org/10.3390/cells11152345
Int J Biol Macromol. 2022 Aug 07. pii: S0141-8130(22)01696-8. [Epub ahead of print]

Find new channel for overcoming chemoresistance in cancers: Role of stem cells-derived exosomal microRNAs.

Ghufran Mohammed Hussein, Shaimaa M Mohammed, Mohammed Faris, Asseal Mohammed, Mohammed Jawad Kadhim, Sura A Awadh, Wafaa H Ajam, Abduladheem Turki Jalil.

  Chemoresistance has remained a significant concern in tumor recurrence and elevated cancer-related mortalities. A deep insight into mechanisms by which cancerous cells resist administered drugs can pave the way to overcome chemotherapy-induced cell death and develop novel procedures to rescue patients. Regarding accumulated data, stem cell-derived exosomal microRNAs (miRNAs) can be deemed a novel and promising method to overcome chemoresistance. It seems exosomal miRNAs play a dual role in the cancer microenvironment. On the one hand, as a messenger, they are transferred between donor and recipient cells contributing to cancer chemoresistance. On the other hand, stem cell-derived exosomal miRNA significantly restrains tumorigenesis and inhibits or alleviates drug resistance in the tumor niche. Hence, our purpose in this review evaluating the roles of stem cells-derived exosomal microRNAs in overcoming chemoresistance in tumors.

Keywords:  Cancer; Chemoresistance; Exosomal microRNAs; Exosomes; microRNAs

DOI:  https://doi.org/10.1016/j.ijbiomac.2022.07.253
Int J Mol Sci. 2022 Aug 02. pii: 8597. [Epub ahead of print]23(15):

A Liquid Biopsy in Bladder Cancer-The Current Landscape in Urinary Biomarkers.

Milena Matuszczak, Adam Kiljańczyk, Maciej Salagierski.

  The non-muscle invasive bladder cancer tends to recur and progress. Therefore, it requires frequent follow-ups, generating costs and making it one of the most expensive neoplasms. Considering the expensive and invasive character of the current gold-standard diagnostic procedure, white-light cystoscopy, efforts to find an alternative method are ongoing. Although the last decade has seen significant advancements in urinary biomarker tests (UBTs) for bladder cancer, international guidelines have not recommended them. Currently, the paramount urgency is to find and validate the test with the best specificity and sensitivity, which would allow for the optimizing of diagnosis, prognosis, and a treatment plan. This review aims to summarise the up-to-date state of knowledge relating to UBTs and new developments in the detection, prognosis, and surveillance of bladder cancer and their potential applications in clinical practice.

Keywords:  biological fluids; bladder cancer; cancer biomarkers; cell-free DNA; cfDNA; diagnosis; exosomes; extracellular vesicles; liquid biopsy; personalized therapy; precision oncology; prognosis; tumour markers

DOI:  https://doi.org/10.3390/ijms23158597
Cancers (Basel). 2022 Jul 30. pii: 3719. [Epub ahead of print]14(15):

Protein Panel of Serum-Derived Small Extracellular Vesicles for the Screening and Diagnosis of Epithelial Ovarian Cancer.

Huiling Lai, Yunyun Guo, Liming Tian, Linxiang Wu, Xiaohui Li, Zunxian Yang, Shuqin Chen, Yufeng Ren, Shasha He, Weipeng He, Guofen Yang.

  Although ovarian cancer, a gynecological malignancy, has the highest fatality rate, it still lacks highly specific biomarkers, and the differential diagnosis of ovarian masses remains difficult to determine for gynecologists. Our study aimed to obtain ovarian cancer-specific protein candidates from the circulating small extracellular vesicles (sEVs) and develop a protein panel for ovarian cancer screening and differential diagnosis of ovarian masses. In our study, sEVs derived from the serum of healthy controls and patients with cystadenoma and ovarian cancer were investigated to obtain a cancer-specific proteomic profile. In a discovery cohort, 1119 proteins were identified, and significant differences in the protein profiles of EVs were observed among groups. Then, 23 differentially expressed proteins were assessed using the parallel reaction monitoring in a validation cohort. Through univariate and multivariate logistic regression analyses, a novel model comprising three proteins (fibrinogen gamma gene (FGG), mucin 16 (MUC16), and apolipoprotein (APOA4)) was established to screen patients with ovarian cancer. This model exhibited an area under the receiver operating characteristic curve (AUC) of 0.936 (95% CI, 0.888-0.984) with 92.0% sensitivity and 82.9% specificity. Another panel comprising serum CA125, sEV-APOA4, and sEV-CD5L showed excellent performance (AUC 0.945 (95% CI, 0.890-1.000), sensitivity of 88.0%, specificity of 93.3%, and accuracy of 89.2%) to distinguish malignancy from benign ovarian masses. Altogether, our study provided a proteomic signature of circulating sEVs in ovarian cancer. The diagnostic proteomic panel may complement current clinical diagnostic measures for screening ovarian cancer in the general population and the differential diagnosis of ovarian masses.

Keywords:  differential diagnosis; extracellular vesicle; ovarian cancer; ovarian cancer screening; proteomics

DOI:  https://doi.org/10.3390/cancers14153719
Int J Mol Sci. 2022 Aug 01. pii: 8544. [Epub ahead of print]23(15):

Detailed Characterization of Small Extracellular Vesicles from Different Cell Types Based on Tetraspanin Composition by ExoView R100 Platform.

Kai Breitwieser, Leon F Koch, Tobias Tertel, Eva Proestler, Luisa D Burgers, Christoph Lipps, James Adjaye, Robert Fürst, Bernd Giebel, Meike J Saul.

  Small extracellular vesicles (sEV) hold enormous potential as biomarkers, drug carriers, and therapeutic agents. However, due to previous limitations in the phenotypic characterization of sEV at the single vesicle level, knowledge of cell type-specific sEV signatures remains sparse. With the introduction of next-generation sEV analysis devices, such as the single-particle interferometric reflectance imaging sensor (SP-IRIS)-based ExoView R100 platform, single sEV analyses are now possible. While the tetraspanins CD9, CD63, and CD81 were generally considered pan-sEV markers, it became clear that sEV of different cell types contain several combinations and amounts of these proteins on their surfaces. To gain better insight into the complexity and heterogeneity of sEV, we used the ExoView R100 platform to analyze the CD9/CD63/CD81 phenotype of sEV released by different cell types at a single sEV level. We demonstrated that these surface markers are sufficient to distinguish cell-type-specific sEV phenotypes. Furthermore, we recognized that tetraspanin composition in some sEV populations does not follow a random pattern. Notably, the tetraspanin distribution of sEV derived from mesenchymal stem cells (MSCs) alters depending on cell culture conditions. Overall, our data provide an overview of the cell-specific characteristics of sEV populations, which will increase the understanding of sEV physiology and improve the development of new sEV-based therapeutic approaches.

Keywords:  characterization; phenotype; small extracellular vesicles; tetraspanins

DOI:  https://doi.org/10.3390/ijms23158544
ACS Nano. 2022 Aug 12.

Biosynthetic Dendritic Cell-Exocytosed Aggregation-Induced Emission Nanoparticles for Synergistic Photodynamic Immunotherapy.

Hongmei Cao, Heqi Gao, Lanxing Wang, Yuanqiu Cheng, Xiaoli Wu, Xiaohong Shen, Hang Wang, Zhen Wang, Panpan Zhan, Jianfeng Liu, Zongjin Li, Deling Kong, Yang Shi, Dan Ding, Yuebing Wang.

  Dendritic cell (DC)-derived small extracellular vesicles (DEVs) are recognized as a highly promising alternative to DC vaccines; however, the clinical testing of DEV-based immunotherapy has shown limited therapeutic efficacy. Herein, we develop a straightforward strategy in which DCs serve as a cell reactor to exocytose high-efficient DEV-mimicking aggregation-induced emission (AIE) nanoparticles (DEV-AIE NPs) at a scaled-up yield for synergistic photodynamic immunotherapy. Exocytosed DEV-AIE NPs inherit not only the immune-modulation proteins from parental DCs, enabling T cell activation, but also the loaded AIE-photosensitizer MBPN-TCyP, inducing superior immunogenic cell death (ICD) by selectively accumulating in the mitochondria of tumor cells. Eventually, DEV-AIE synergistic photodynamic immunotherapy elicits dramatic immune responses and efficient eradication of primary tumors, distant tumors, and tumor metastases. In addition, cancer stem cells (CSCs) in 4T1 and CT26 solid tumors were significantly inhibited by the immune functional DEV-AIE NPs. Our work presents a facile method for the cellular generation of EV-biomimetic NPs and demonstrates that the integration of DEVs and AIE photosensitizers is a powerful direction for the production of clinical anticancer nanovaccines.

Keywords:  aggregation-induced emission; dendritic cells; exocytosis; extracellular vesicles; immunotherapy

DOI:  https://doi.org/10.1021/acsnano.2c03597
Cells. 2022 Jul 27. pii: 2309. [Epub ahead of print]11(15):

Selective Internal Radiotherapy Changes the Immune Profiles of Extracellular Vesicles and Their Immune Origin in Patients with Inoperable Cholangiocarcinoma.

Florian Haag, Anjana Manikkam, Daniel Kraft, Caroline Bär, Vanessa Wilke, Aleksander J Nowak, Jessica Bertrand, Jazan Omari, Maciej Pech, Severin Gylstorff, Borna Relja.

  The incidence of cholangiocellular carcinoma (CCA) is rising worldwide. As there are no specific early symptoms or specific markers of CCA, it is often diagnosed in later inoperable stages. Accumulating evidence underlines the importance of radiation therapy in the induction of antitumor immunity. The surface protein composition on extracellular vesicles (EVs) relates to originating cells and thus may play a role in vesicle function. We assessed immune profiles of EVs and their immune origin in patients with inoperable CCA prior and after selective internal radiotherapy (SIRT). A total of 47 CCA patients receiving SIRT and 12 healthy volunteers (HV) were included. Blood was withdrawn before therapy (pre T) and after T. EVs were purified from plasma by cluster of differentiation (CD)9-, CD63-, and CD81-immunobead isolation. To detect differently abundant surface markers, dynamic range and EVs input quality were assessed. A total of 37 EVs surface markers were measured by flow cytometry and correlated either with the administered activity dose (MBq) or with the interval until death (month). EVs phenotyping identified lymphocytes, B cells, NK cells, platelets, endothelial cells, leukocyte activation, B cell activation, T and B cell adhesion markers, stem/progenitor cells, and antigen-presenting cells (APC) as EVs-parenteral cells. CD4 and CD8 significantly declined, while other markers significantly increased in CCA patients pre T vs. HV. Platelets-deriving EVs significantly decreased, normalizing to levels of HV but still significantly increasing vs. HV post SIRT. B cells-deriving EVs significantly increased pre T vs. HV, positively correlating with administered activity dose. MHCII and CD40 EVs significantly increased pre SIRT and negatively correlated with administered activity dose, while EVs from antigen presenting cells and CD49e pre SIRT positively correlated with survival time after therapy. Increased levels of CD24 and CD44 in cancer pre T were significantly decreased post T. Among the heterogeneity of EVs that was demonstrated, in particular, B cells-deriving, MHCII, and CD40 positive or APC-deriving EVs need to be further studied for their diagnostic or prognostic relevance in clinical scenarios.

Keywords:  CCA; EVs; MHC; biomarker; diagnosis; prognosis

DOI:  https://doi.org/10.3390/cells11152309
Clin Transl Oncol. 2022 Aug 10.

Diagnostic value of exosomes in patients with liver cancer: a systematic review.

Jusong Liu, Pan Xiao, Wenxue Jiang, Yuhan Wang, Yuanshuai Huang.

  BACKGROUND: Liver cancer is a disease with high morbidity and mortality. More and more studies have shown that exosomes can be used as biomarkers for the diagnosis of liver cancer, but their diagnostic accuracy is still unclear. Therefore, this meta-analysis summarizes various studies on the diagnostic value of exosomes for liver cancer.METHODS: A comprehensive search was carried out based on the set search terms in PubMed, Web of Science and Wiley until April 1, 2022. All statistical analyses were performed by STATA 17 statistical software and Review Manager 5.4. Quality Assessment for Studies of Diagnostic Accuracy 2 tool was applied to evaluate the quality of included articles. Random effects model was used to calculate various diagnostic indicators.
RESULTS: A total of 47 studies were included in this meta-analysis. The number of participants was 3196. The combined sensitivity, specificity and the area under the curve with 95% confidence intervals (95% CI) were, respectively 0.80 (0.75-0.84), 0.83 (0.79-0.87), 0.89 (0.85-0.91).
CONCLUSIONS: This meta-analysis shows that exosomes have good diagnostic accuracy for liver cancer and can be used as an effective biomarker for the diagnosis of liver cancer.

Keywords:  Biomarker; Diagnosis; Exosomes; Liver cancer; Meta-analysis

DOI:  https://doi.org/10.1007/s12094-022-02906-8
Talanta. 2022 Jul 31. pii: S0039-9140(22)00562-8. [Epub ahead of print]251 123766

Microfluidic amperometric immunosensor based on porous nanomaterial towards claudin7 determination for colorectal cancer diagnosis.

Francisco G Ortega, Germán E Gomez, Chiara Boni, Inés Cañas García, Carmen Garrido Navas, Richard F D'vries, María Pilar Molina Vallejos, María José Serrano, Germán A Messina, José Expósito Hernández, Martín A Fernández-Baldo.

  In this work, we present a microfluidic amperometric immunosensor for cancer biomarker claudin7 (CLD7) determination in circulating extracellular vesicles (EVs) as well as its validation in colorectal cancer (CC) patients. The device is based on synthetized nanosized MIL-125-NH2 particles, covalently anchored to the central channel of the microfluidic immunosensor. This nanomaterial was employed as efficient platform for anti-CLD7 monoclonal antibodies immobilization for specifically recognize and capture CLD7 in EVs samples. Afterwards, the amount of this trapped CLD7 was quantified by HRP-conjugated anti-CLD7-antibody. HRP reacted with its enzymatic substrate in a redox process which resulted in the appearance of a current whose magnitude was directly proportional to the level of CLD7 in the sample. This immunosensor, under optimum conditions, gave the limit of detection for CLD7 of 0.1 pg mL-1, with a wide linear range from 2 to 1000 pg mL-1. The results reported herein open up the use of porous open framework platforms for sensing applications for biomedicine and diagnosis.

Keywords:  Biosensor; Cancer biomarker; Electrochemical; Extracellular vesicles; MOF platform; Nanomaterial

DOI:  https://doi.org/10.1016/j.talanta.2022.123766
Biosci Trends. 2022 Aug 07.

Hepatic stellate cell exosome-derived circWDR25 promotes the progression of hepatocellular carcinoma via the miRNA-4474-3P-ALOX-15 and EMT axes.

Lei Liu, Rui Liao, Zhongjun Wu, Chengyou Du, Yu You, Keting Que, Yuxin Duan, Kunli Yin, Wentao Ye.

  Recently, the emerging role of circular RNAs (circRNAs) in tumor development and progression has been a topic of great interest. Nevertheless, the effects of hepatic stellate cell (HSC)-derived exosomes in hepatocellular carcinoma (HCC) remain unclear. Here, we aim to explore the potential effect of HSC exosome-derived circWDR25 on the aggressiveness of HCC. Firstly, a microarray analysis of circRNAs was performed to profile and identify the differentially expressed circRNAs derived from HSC exosomes activated by HCC cells. Subsequently, the roles of circWDR25 in HCC tumor growth and aggressiveness were confirmed through in vitro and in vivo functional experiments. Moreover, RNA pull-down, dual-luciferase reporter assays, and fluorescent in situ hybridization (FISH) were performed to determine interactions in the circWDR25-miR-4474-3p-ALOX15 loop. Immunohistochemical analysis was also performed on a microarray of HCC tissues and peritumoral tissues. We found that overexpressed peritumoral circWDR25 was associated with survival and recurrence in patients with HCC and promoted the progression of HCC cells both in vitro and in vivo. Mechanistically, both exogenous and HSC exosomal-derived circWDR25 regulated the expression of ALOX15 by sponging miR-4474-3p and ultimately inducing an epithelial-to-mesenchymal transition (EMT) in HCC cells. Moreover, exogenous and HSC exosomal-derived circWDR25 promoted the expression of CTLA-4 in HSCs and PD-L1 in HCC cells. In conclusion, circWDR25 facilitated HCC cell proliferation and invasion via the circWDR25/miR-4474-3p/ALOX15 and EMT axes and it promoted the expression of CTLA-4 in HSCs and PD-L1 in HCC cells, thus providing insights into the mechanism of tumor aggressiveness mediated by HSC-derived exosomal circWDR25.

Keywords:  circular RNA; extracellular vesicles; liver cancer; mesenchymal cells; prognosis

DOI:  https://doi.org/10.5582/bst.2022.01281
Int J Mol Sci. 2022 Jul 28. pii: 8351. [Epub ahead of print]23(15):

Spotlight on Exosomal Non-Coding RNAs in Breast Cancer: An In Silico Analysis to Identify Potential lncRNA/circRNA-miRNA-Target Axis.

Ohanes Ashekyan, Samira Abdallah, Ayman Al Shoukari, Ghada Chamandi, Hayat Choubassy, Abdul Rahman S Itani, Nisreen Alwan, Rihab Nasr.

  Breast cancer (BC) has recently become the most common cancer type worldwide, with metastatic disease being the main reason for disease mortality. This has brought about strategies for early detection, especially the utilization of minimally invasive biomarkers found in various bodily fluids. Exosomes have been proposed as novel extracellular vesicles, readily detectable in bodily fluids, secreted from BC-cells or BC-tumor microenvironment cells, and capable of conferring cellular signals over long distances via various cargo molecules. This cargo is composed of different biomolecules, among which are the novel non-coding genome products, such as microRNAs (miRNAs), long non-coding RNAs (lncRNAs), and the recently discovered circular RNA (circRNA), all of which were found to be implicated in BC pathology. In this review, the diverse roles of the ncRNA cargo of BC-derived exosomes will be discussed, shedding light on their primarily oncogenic and additionally tumor suppressor roles at different levels of BC tumor progression, and drug sensitivity/resistance, along with presenting their diagnostic, prognostic, and predictive biomarker potential. Finally, benefiting from the miRNA sponging mechanism of action of lncRNAs and circRNAs, we established an experimentally validated breast cancer exosomal non-coding RNAs-regulated target gene axis from already published exosomal ncRNAs in BC. The resulting genes, pathways, gene ontology (GO) terms, and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis could be a starting point to better understand BC and may pave the way for the development of novel diagnostic and prognostic biomarkers and therapeutics.

Keywords:  biomarkers; breast cancer; circular RNAs; exosomes; long non-coding RNA; microRNAs; non-coding RNA

DOI:  https://doi.org/10.3390/ijms23158351
Biotechniques. 2022 Aug 10.

A cryostat-based frozen section method to increase the yield of extracellular vesicles extracted from different tissues.

Shuangzhu Yang, Weiwei Qin, Xiangji Li, Qingdong Guo, Yuan Wang, Jing Chen, Hongmei Li, Shutian Zhang, Li Min.

  Extracellular vesicles (EVs) are small vesicles mediating intercellular communications that have been widely used in disease diagnosis. Extracting EVs from tissues is of great importance, but current approaches are finite and the EV yield is limited. Here, the authors introduced a new method to increase EV yield based on frozen sectioning. With a standardized, semiautomated tissue-slicing procedure in a cryostat, the authors successfully isolated EVs from hearts, kidneys and stomachs. The morphology, size distribution and purity of those isolated EVs were evaluated. Additionally, compared with the traditional scalpel section method, they confirmed the higher yield of tissue-derived EVs with the cryostat-based method. The authors believe that the new method they developed would largely facilitate the research and clinical application of EVs.

Keywords:  extracellular vesicles; size exclusion chromatography; tissues; ultracentrifugation; ultrafiltration

DOI:  https://doi.org/10.2144/btn-2022-0035
Anticancer Drugs. 2022 Aug 09.

Identification of exosomal circRNA CD226 as a potent driver of nonsmall cell lung cancer through miR-1224-3p/high mobility group AT-hook 2 axis.

Lihong Peng, Yunpeng Cao, Chao Hu.

Circular RNAs (circRNAs) are crucial for the pathogenesis of nonsmall lung cancer (NSCLC). Here, we set out to unravel the precise function of circRNA CD226 (circCD226) in NSCLC pathogenesis. The exosomes from serum specimens were observed by transmission electron microscopy. CircCD226, miR-1224-3p and high mobility group AT-hook 2 (HMGA2) were quantified by qRT-PCR, western blot and immunohistochemistry. Actinomycin D and Ribonuclease (RNase) R treatments and subcellular localization assay were used for circCD226 characterization. Cell viability, proliferation, migration, invasion and sphere formation abilities were gauged by CCK-8, EDU, wound-healing, transwell and sphere formation assays, respectively. Directed relationships among circCD226, miR-1224-3p and HMGA2 were examined by RNA pull-down, dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. The abundance of circCD226 was elevated in serum exosomes, tissues and cells of NSCLC. NSCLC serum exosomes enhanced NSCLC cell proliferation, migration, invasion and stemness. Loss of circCD226 impeded cell proliferation, migration, invasion and stemness in vitro, as well as tumor growth in vivo. Mechanistically, circCD226 sponged miR-1224-3p, and miR-1224-3p targeted HMGA2. CircCD226 involved the posttranscriptional regulation of HMGA2 through miR-1224-3p. Moreover, the miR-1224-3p/HMGA2 axis was identified as a functionally downstream effector of circCD226 in regulating NSCLC cell behaviors. Our study identifies circCD226 as a potential driver in NSCLC development depending on the regulation of miR-1224-3p/HMGA2 axis.

DOI: https://doi.org/10.1097/CAD.0000000000001357
Cell Prolif. 2022 Aug 09. e13312

Human platelet lysate-derived extracellular vesicles enhance angiogenesis through miR-126.

Antonella Bordin, Maila Chirivì, Francesca Pagano, Marika Milan, Marco Iuliano, Eleonora Scaccia, Orazio Fortunato, Giorgio Mangino, Xhulio Dhori, Elisabetta De Marinis, Alessandra D'Amico, Selenia Miglietta, Vittorio Picchio, Roberto Rizzi, Giovanna Romeo, Fabio Pulcinelli, Isotta Chimenti, Giacomo Frati, Elena De Falco.

OBJECTIVES: Extracellular vesicles (EVs) are key biological mediators of several physiological functions within the cell microenvironment. Platelets are the most abundant source of EVs in the blood. Similarly, platelet lysate (PL), the best platelet derivative and angiogenic performer for regenerative purposes, is enriched of EVs, but their role is still too poorly discovered to be suitably exploited. Here, we explored the contribution of the EVs in PL, by investigating the angiogenic features extrapolated from that possessed by PL.METHODS: We tested angiogenic ability and molecular cargo in 3D bioprinted models and by RNA sequencing analysis of PL-derived EVs.
RESULTS: A subset of small vesicles is highly represented in PL. The EVs do not retain aggregation ability, preserving a low redox state in human umbilical vein endothelial cells (HUVECs) and increasing the angiogenic tubularly-like structures in 3D endothelial bioprinted constructs. EVs resembled the miRNome profile of PL, mainly enriched with small RNAs and a high amount of miR-126, the most abundant angiogenic miRNA in platelets. The transfer of miR-126 by EVs in HUVEC after the in vitro inhibition of the endogenous form, restored angiogenesis, without involving VEGF as a downstream target in this system.
CONCLUSION: PL is a biological source of available EVs with angiogenic effects involving a miRNAs-based cargo. These properties can be exploited for targeted molecular/biological manipulation of PL, by potentially developing a product exclusively manufactured of EVs.

DOI: https://doi.org/10.1111/cpr.13312
Cell Oncol (Dordr). 2022 Aug 11.

Therapeutic exosomes loaded with SERPINA5 attenuated endometrial cancer cell migration via the integrin β1/FAK signaling pathway.

Yunfeng Song, Lei Ye, Yuan Tan, Huan Tong, Zeheng Lv, Xiaoping Wan, Yiran Li.

  BACKGROUND: Metastasis is still the major cause of endometrial cancer (EC)-related death. Because of their biological function and regenerative properties, exosomes have been applied to therapeutic regimens. SERPINA5 expression is downregulated in several tumors and linked to tumor cell migration and invasion. However, the expression and biological functions of SERPINA5 in EC remain unclear.METHODS: The levels of SERPINA5 in plasma exosomes were determined with ELISAs. SERPINA5 expression in EC and its relationship with survival outcomes were analyzed using the TCGA database and clinical EC tissue samples. The effect of SERPINA5 overexpression or exosomal SERPINA5 on EC metastasis was examined by cell migration and invasion assays in vitro. Mechanistically, overexpression of SERPINA5 or high exosomal SERPINA5 levels mediated the regulation of the integrin β1/FAK signaling pathway in EC cell lines. The therapeutic effect of exosomal SERPINA5 was determined with xenograft models.
RESULTS: This study revealed that the level of exosomal SERPINA5 was increased in the circulating plasma of EC patients. In addition, the expression of SERPINA5 was decreased in EC patients with distant metastasis, and low expression of SERPINA5 indicated worse survival. In addition, SERPINA5 was elevated in normal tissues adjacent to EC tumors. Moreover, overexpression of SERPINA5 inhibited metastatic potential of EC cell lines in vitro. Furthermore, SERPINA5 loaded on secreted exosomes reduced the metastatic ability of EC cells. Notably, overexpression of SERPINA5 or high exosomal SERPINA5 levels suppressed EC metastatic potential by suppressing integrin β1/FAK signaling pathway activation. Finally, exosomal SERPINA5 impeded tumor growth and metastasis in xenograft models.
CONCLUSIONS: Our findings revealed that a low level of SERPINA5 expression indicated poor survival outcomes in EC and that exogenous SERPINA5 loading of exosomes may be a novel therapeutic strategy for metastatic EC.

Keywords:  Endometrial cancer; Exosome; FAK; SERPINA5

DOI:  https://doi.org/10.1007/s13402-022-00687-4
Cancers (Basel). 2022 Jul 31. pii: 3731. [Epub ahead of print]14(15):

The Hepatic Pre-Metastatic Niche.

Benjamin Ormseth, Amblessed Onuma, Hongji Zhang, Allan Tsung.

  Primary tumors can communicate with the liver to establish a microenvironment that favors metastatic colonization prior to dissemination, forming what is termed the "pre-metastatic niche" (PMN). Through diverse signaling mechanisms, distant malignancies can both influence hepatic cells directly as well as recruit immune cells into the PMN. The result is a set of changes within the hepatic tissue that increase susceptibility of tumor cell invasion and outgrowth upon dissemination. Thus, the PMN offers a novel step in the traditional metastatic cascade that could offer opportunities for clinical intervention. The involved signaling molecules also offer promise as biomarkers. Ultimately, while the existence of the hepatic PMN is well-established, continued research effort and use of innovative models are required to reach a functional knowledge of PMN mechanisms that can be further targeted.

Keywords:  ECM remodeling; bone-marrow-derived cells; exosomes; immunosuppression; liver; metastasis; pre-metastatic niche

DOI:  https://doi.org/10.3390/cancers14153731
Tissue Eng Part C Methods. 2022 Aug 12.

A systematic review of EV-piRNA in human body fluid and its role in disease progression.

Tuan Xin Goh, Sik Loo Tan, Margaret M Roebuck, Seow-Hui Teo, Tunku Kamarul.

The state of host cells is reflected in the cargo carried by their extracellular vesicles (EVs). This makes EV a potential source of biomarkers for human diseases. Piwi-interacting RNA (piRNA) regulates gene expression through epigenetic regulation and post-transcriptional gene silencing. Thus, piRNA profiles in EVs derived from human clinical samples could identify markers that characterize disease stages, and unveil their roles in disease pathology. This review aimed to report the expression profiles of EV-derived piRNA (EV-piRNA) in various human samples, as well as their role in each pathology. A systematic review was conducted to collate the findings of human EV-piRNA from original research articles published in indexed scientific journals up to 16th Feb 2022. Article searches were performed in PubMed, Web of Science, and Scopus databases, using a combination of keywords including 'EV' and 'piRNA'. A total of 775 non-redundant original articles were identified. After subjecting articles to inclusion and exclusion criteria, 34 articles were accepted for this review. The piRNA expression levels among the small RNA profiles of human-derived EVs range from 0.09% to 43.84%, with the lowest expression level reported in urine-derived EVs and the highest percentage in plasma-derived EVs. Differentially expressed EV-piRNAs have been identified in patients with specific disease conditions compared to their counterparts (healthy control), suggesting an association between piRNA and progression in various diseases. Seven articles identified piRNA putative target genes and/or the pathway enrichment of piRNA target genes, and one study demonstrated a direct tole of piRNA candidates in disease pathology. In conclusion, EV-piRNA has been isolated successfully from various human body fluids. EV-piRNA is a new research niche in human disease pathology. The expression profiles of EV-piRNA in various tissue types and disease conditions remain largely unexplored. Furthermore, there is currently a lack of guidelines on piRNA bioinformatics analysis, which could lead to inconsistent results and thus hinder the progression of piRNA discoveries. Lastly, the lack of published scientific evidence on the role of EV-piRNA supports the need for future research to focus on the functional analysis of EV-piRNA as part of the route in piRNA discoveries.

DOI: https://doi.org/10.1089/ten.TEC.2022.0092