bims-exocan 2022-08-21 papers

bims-exocan

Biomed News

on Exosomes roles in cancer

Issue of 2022–08–21
seventeen papers selected by
Muhammad Rizwan, COMSATS University

Role of exosomes in tumor growth, chemoresistance and immunity: state-of-the-art.
Cancer-Derived Small Extracellular Vesicles PICKER.
Exosomes: Tools and targets for cancer diagnosis, prognosis, and therapy.
Exosome-transmitted miR-3124-5p promotes cholangiocarcinoma development via targeting GDF11.
Characterization of extracellular vesicles by flow cytometry: Challenges and promises.
Emerging micro-nanotechnologies for extracellular vesicles in immuno-oncology: from target specific isolations to immunomodulation.
Cancer Stem Cell Derived Extracellular Vesicles with Self-Functionalized 3D Nanosensor for Real-Time Cancer Diagnosis: Eliminating the Roadblocks in Liquid Biopsy.
α2,6-Sialylation promotes hepatocellular carcinoma cells migration and invasion via enhancement of nSmase2-mediated exosomal miRNA sorting.
miR-222-3p-containing macrophage-derived extracellular vesicles confer gemcitabine resistance via TSC1-mediated mTOR/AKT/PI3K pathway in pancreatic cancer.
Extracellular Vesicles and Circulating Tumour Cells - complementary liquid biopsies or standalone concepts?
Tumor associated macrophages-derived exosomes facilitate hepatocellular carcinoma malignance by transferring lncMMPA to tumor cells and activating glycolysis pathway.
Cancer cell-derived exosomal LINC00313 induces M2 macrophage differentiation in non-small cell lung cancer.
Decrease of circARID1A retards glioblastoma invasion by modulating miR-370-3p/ TGFBR2 pathway.
Exosomal miR-155-5p derived from glioma stem-like cells promotes mesenchymal transition via targeting ACOT12.
How I Use It: The Exosome Diagnostics (EPI) prostate cancer biomarker utility in urology and primary care.
Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts.
[Release of Exosomes Derived from Leukocyte-Depleted Red Cell Suspension and Its Regulation on Hematological Tumor Cells].

J Drug Target. 2022 Aug 16. 1-35

Role of exosomes in tumor growth, chemoresistance and immunity: state-of-the-art.

Seyedeh Maryam Hosseinikhah, Fatemeh Gheybi, Seyedeh Alia Moosavian, Mohammad-Ali Shahbazi, Mahmoud Reza Jaafari, Mika Sillanpää, Prashant Kesharwani, Seyedeh Hoda Alavizadeh, Amirhossein Sahebkar.

  Cancer is one of the most lethal diseases, and limited available treatment options contribute to its high mortality rate. Exosomes are considered membrane-bound nanovesicles that include different molecules such as lipids, proteins, and nucleic acids. Virtually most cells could release exosomes via exocytosis in physiological and pathological conditions. Tumor-derived exosomes (TDEs) play essential roles in tumorigenesis, proliferation, progression, metastasis, immune escape, and chemoresistance by transferring functional biological cargos, triggering different autocrine, and paracrine signaling cascades. Due to their antigen-presenting properties, exosomes are widely used as biomarkers and drug carriers and have a prominent role in cancer immunotherapy. They offer various advantages in carrier systems (e.g., in chemotherapy, siRNA, and miRNA), delivery of diagnostic agents owing to their stability, loading of hydrophobic and hydrophilic agents, and drug targeting. Novel exosomes-based carriers can be generated as intelligent systems using various sources and crosslinking chemistry extracellular vesicles (EVs). Exosomes studded with targeting ligands, including peptides, can impart in targeted delivery of cargos to tumor cells. In this review, we comprehensively summarized the important role of tumor-derived exosomes in dictating cancer pathogenesis and resistance to therapy. We have therefore, investigated in further detail the pivotal role of tumor-derived exosomes in targeting various cancer cells and their applications, and prospects in cancer therapy and diagnosis. Additionally, we have implicated the potential utility and significance of tumor exosomes-based nanoparticles as an efficient and novel therapeutic carrier and their applications in treating advanced cancers.

Keywords:  Chemoresistance; Diagnosis; Drug delivery; Immunomodulation; Tumor-derived exosomes

DOI:  https://doi.org/10.1080/1061186X.2022.2114000
Anal Chem. 2022 Aug 18.

Cancer-Derived Small Extracellular Vesicles PICKER.

Xiaohui Chen, Yun Deng, Ruyan Niu, Zixin Sun, Alya Batool, Liu Wang, Chong Zhang, Ningyu Ma, Qingtang Yang, Guoxiang Liu, Jichun Yang, Yang Luo.

Cancer-derived small extracellular vesicles (csEVs) play critical roles in the genesis and development of various cancers. However, accurate detection of low-abundance csEVs remains particularly challenging due to the complex clinical sample composition. In the present study, we constructed a Programmable Isothermal Cascade Keen Enzyme-free Reporter (PICKER) for the reliable detection and acquisition of the relative abundance of csEVs in total sEVs (tsEVs) by integrating dual-aptamer recognition (cancer-specific protein EpCAM and tetraspanin protein CD63) with a catalytic hairpin assembly (CHA) amplification. By employing this strategy, we were able to achieve a detection limit of 420 particles/μL csEVs. Particularly, we proposed a novel particle ratio index of csEV against tsEV (PRcsEV/tsEV) to greatly eliminate errors from inconsistent centrifugation, which was calculated from the fluorescence ratio produced by csEVs and tsEVs. The PICKER showed a 1/10,000 discrimination capability by successfully picking out 1.0 × 103 csEV from 1.0 × 107 tsEV per microliter. We also found that the PRcsEV/tsEV value increased proportional to the stages of breast cancer by analyzing EVs from clinical patients' plasma. Taken together, we established a PICKER strategy capable of accurately discriminating csEVs, and the proposed PRcsEV/tsEV had been proven a potential indicator of breast cancer staging, paving the way toward facilitating cancer diagnosis and precision therapeutics.

DOI: https://doi.org/10.1021/acs.analchem.2c01683
Cancer Lett. 2022 Aug 10. pii: S0304-3835(22)00349-4. [Epub ahead of print] 215865

Exosomes: Tools and targets for cancer diagnosis, prognosis, and therapy.

Seema Singh.

Exosomes are a subset of extracellular vesicles (EVs) secreted by most eukaryotic cells. These lipid-bilayer membrane structures are typically in the size range of 40-160 nanometers in diameter and are originated via an endosomal pathway. Exosomes were discovered nearly four decades ago and were initially thought to serve as cellular garbage bags whose sole function was to remove unnecessary material from the cells. About a decade later, exosomes were shown to have a biological function, and since then, it has become clear that they contribute immensely to many aspects of health and disease by facilitating inter-cellular communications. Exosomes carry valuable information from the donor cells, such as cell surface and intracellular proteins, RNA, DNA, metabolites, and other molecular constituents, and deliver it to the recipient cells altering their functions. During the past two decades, exosomes have received significant attention in the field of cancer biology. They are being aggressively studied for their roles in pathogenesis, biomarker development for diagnosis and monitoring, and as therapeutic targets for treatment. More recently, numerous studies have investigated if exosomes could also be used as vehicles to efficiently deliver drugs and other bioactive molecules to cancer cells. The study of exosomes and their clinical applications in cancer continues to evolve and hold great potential to impact cancer management.

DOI: https://doi.org/10.1016/j.canlet.2022.215865
Front Oncol. 2022 ;12 936507

Exosome-transmitted miR-3124-5p promotes cholangiocarcinoma development via targeting GDF11.

Huijie Gao, Zhaobin He, Chao Gao, Naiqing Liu, Zhaoyang Zhang, Weibo Niu, Jun Niu, Cheng Peng.

   Objective: Cholangiocarcinoma (CHOL) is a deadly cancer worldwide with limited available therapies. The aim of this study was to investigate key exosomal miRNAs and their functions in CHOL development.
Methods: Serum exosomes were isolated from patients with CHOL and healthy controls, followed by miRNA sequencing for identifying differentially expressed miRNAs (DEMs) and their functions. Then, the expression of key DEMs was experimentally validated in exosomes from clinical CHOL patients and CHOL cells. The effects of overexpression of key DEMs on CHOL cell migration and proliferation were investigated. A key exosomal DEM miR-3124-5p was identified. The effects of overexpression or knockdown of exosomal miR-3124-5p on the proliferation, migration, and angiogenesis of human umbilical vein endothelial cells (HUVECs) were investigated. Moreover, the function of exosomal miR-3124-5p on tumor growth in vivo was explored.
Results: A total of 632 exosomal DEMs were identified between CHOL and control samples. Target genes of DEMs were significantly enriched in pathways, such as the p53 signaling pathway. miR-3124-5p was upregulated in serum exosomes from CHOL patients and exosomes from CHOL cells, and overexpression of miR-3124-5p promoted RBE cell migration and viability. Moreover, overexpression of exosomal miR-3124-5p promoted the proliferation, migration, and angiogenesis of HUVECs, while knockdown of miR-3124-5p had the opposite effect. miR-3124-5p could target growth differentiation factor 11 (GDF11) and downregulate GDF11 expression. Furthermore, exosomal miR-3124-5p promoted tumor growth in vivo.
Conclusions: Our findings revealed that exosome-encapsulated miR-3124-5p promoted the malignant progression of CHOL by targeting GDF11. Exosomal miR-3124-5p and GDF11 could be promising biomarkers or therapeutic targets for CHOL.

Keywords:  cholangiocarcinoma; exosome; growth differentiation factor 11; miR-3124-5p; miRNA

DOI:  https://doi.org/10.3389/fonc.2022.936507
Micron. 2022 Aug 05. pii: S0968-4328(22)00137-8. [Epub ahead of print]161 103341

Characterization of extracellular vesicles by flow cytometry: Challenges and promises.

Banat Gul, Feryal Syed, Shamim Khan, Abid Iqbal, Iftikhar Ahmad.

  The International Society for Extracellular Vesicles (ISEV) defines the extracellular vesicle (EV) as "the particles naturally released from the cell that are delimited by a lipid bilayer and cannot replicate, i.e. do not contain a functional nucleus". The size (diameter) of EVs ranges in ~30-1000 nm, with peak population at ~ 100 nm. Flow cytometry (FCM) is the most commonly used technique for analysis of EVs. However, accurate characterization, procedure standardization, instrument calibration and results interpretation/validation of EVs is confounded by their complex and variable composition, small size and substantial differences in physiological concentrations. Here, the challenges to and promises of FCM for characterization of EVs are discussed. Specifically, we systematically reviewed the pitfalls of FCM in the detection of (small) EVs and the corresponding strategies for enhancing the sensitivity and resolution of the instrument. The shortcomings and improvement in the overall FCM system are described in terms of reference material for calibration, the collection optics for fluorescence (FL), side scatter (SSC) and forward scatter (FSC) signals and fluidics. This study may provide a comprehensive reference for a brief overview pertaining to the challenges and promises of a modern FCM system for analysis of EVs.

Keywords:  Extracellular vesicles; Flow cytometry; Fluorescence; Light scattering; Liquid biopsy; Reference beads

DOI:  https://doi.org/10.1016/j.micron.2022.103341
Lab Chip. 2022 Aug 18.

Emerging micro-nanotechnologies for extracellular vesicles in immuno-oncology: from target specific isolations to immunomodulation.

Nna-Emeka Onukwugha, Yoon-Tae Kang, Sunitha Nagrath.

Extracellular vesicles (EVs) have been hypothesized to incorporate a variety of crucial roles ranging from intercellular communication to tumor pathogenesis to cancer immunotherapy capabilities. Traditional EV isolation and characterization techniques cannot accurately and with specificity isolate subgroups of EVs, such as tumor-derived extracellular vesicles (TEVs) and immune-cell derived EVs, and are plagued with burdensome steps. To address these pivotal issues, multiplex microfluidic EV isolation/characterization and on-chip EV engineering may be imperative towards developing the next-generation EV-based immunotherapeutics. Henceforth, our aim is to expound the state of the art in EV isolation/characterization techniques and their limitations. Additionally, we seek to elucidate current work on total analytical system based technologies for simultaneous isolation and characterization and to summarize the immunogenic capabilities of EV subgroups, both innate and adaptive. In this review, we discuss recent state-of-art microfluidic/micro-nanotechnology based EV screening methods and EV engineering methods towards therapeutic use of EVs in immune-oncology. By venturing in this field of EV screening and immunotherapies, it is envisioned that transition into clinical settings can become less convoluted for clinicians.

DOI: https://doi.org/10.1039/d2lc00232a
ACS Nano. 2022 Aug 15.

Cancer Stem Cell Derived Extracellular Vesicles with Self-Functionalized 3D Nanosensor for Real-Time Cancer Diagnosis: Eliminating the Roadblocks in Liquid Biopsy.

Rupa Haldavnekar, Krishnan Venkatakrishnan, Bo Tan.

  Liquid biopsy for determining the presence of cancer and the underlying tissue of origin is crucial to overcome the limitations of existing tissue biopsy and imaging-based techniques by capturing critical information from the dynamic tumor heterogeneity. A newly emerging liquid biopsy with extracellular vesicles (EVs) is gaining momentum, but its clinical relevance is in question due to the biological and technical challenges posed by existing technologies. The biological barriers of existing technologies include the inability to generate fundamental details of molecular structure, chemical composition as well as functional variations in EVs by gathering simultaneous information on multiple intra-EV molecules, unavailability of holistic qualitative analysis, in addition to the inability to identify tissue of origin. Technological barriers include reliance on EV isolation with a few labeled biomarkers, resulting in the inability to generate comprehensive information on the disease. A more favorable approach would be to generate holistic information on the disease without the use of labels. Such a marker-free diagnosis is impossible with the existing liquid biopsy due to the unavailability clinically validated cancer stem cells (CSC)-specific markers and dependence of existing technologies on EV isolation, undermining the clinical relevance of EV-based liquid biopsy. Here, CSC EVs were employed as an independent liquid biopsy modality. We hypothesize that tracking the signals of CSCs in peripheral blood with CSC EVs will provide a reliable solution for accurate cancer diagnosis, as CSC are the originators of tumor contributing to tumor heterogeneity. We report nanoengineered 3D sensors of extremely small nano-scaled probes self-functionalized for SERS, enabling integrative molecular and functional profiling of otherwise undetectable CSC EVs. A substantially enhanced SERS and ultralow limit of detection (10 EVs per 10 μL) were achieved. This was attributed to the efficient probe-EV interaction due to the 3D networks of nanoprobes, ensuring simultaneous detection of multiple EV signals. We experimentally demonstrate the crucial role of CSC EVs in cancer diagnosis. We then completed a pilot validation of this modality for cancer detection as well as for identification of the tissue of origin. An artificial neural network distinguished cancer from noncancer with 100% sensitivity and 100% specificity for three hard to detect cancers (breast, lung, and colorectal cancer). Binary classification to distinguish one tissue of origin against all other achieved 100% accuracy, while simultaneous identification of all three tissues of origin with multiclass classification achieved up to 79% accuracy. This noninvasive tool may complement existing cancer diagnostics, treatment monitoring as well as longitudinal disease monitoring by validation with a large cohort of clinical samples.

Keywords:  SERS; cancer stem cells; extracellular vesicles; liquid biopsy; machine learning; nanosensor

DOI:  https://doi.org/10.1021/acsnano.2c02971
J Physiol Biochem. 2022 Aug 19.

α2,6-Sialylation promotes hepatocellular carcinoma cells migration and invasion via enhancement of nSmase2-mediated exosomal miRNA sorting.

Liping Wang, Xixi Chen, Fanxu Meng, Tianmiao Huang, Shujing Wang, Zhichao Zheng, Guoliang Zheng, Wenli Li, Jianing Zhang, Yubo Liu.

  Exosomes have a critical role in the intercellular communication and metastatic progression of hepatocellular carcinoma (HCC). Recently, our group showed that α2, 6-sialylation played an important role in the proliferation- and migration-promoting effects of cancer-derived exosomes. However, the molecular basis remains elusive. In this study, the mechanism of α2, 6-sialylation-mediated specific microRNAs (miRNA) sorting into exosomes was illustrated. We performed miRNA profiling analysis to compare exosomes from HCC cell lines that differ only in α2, 6-sialylation status. A total of 388 differentially distributed miRNAs were identified in wild-type and β-galactoside α2, 6-sialyltransferase I (ST6Gal-I) knockdown MHCC-97H cells-derived exosomes. Neutral sphingomyelinase-2 (nSmase2), an important regulator mediating the sorting of exosomal miRNAs, was found to be a target of ST6Gal-I. The reduction of α2, 6-sialylation could impair the activity of nSmase2, as well as the nSmase2-dependent exosomal miRNAs sorting. This α2,6-sialylation-dependent sorting exerted an augmentation of motility on recipient HCC cells. Our data further demonstrated that α2,6-sialylation-mediated sorting of exosomal miR-100-5p promoted the migration and invasion of recipient HepG2 cells via the PI3K/AKT signaling pathway. The cellular metastasis-related gene CLDN11 was confirmed as a direct target of exosomal miR-100-5p, which elevated the mobility of recipient HCC cells. In conclusion, our results showed that α2,6-sialylation modulates nSmase2-dependent exosomal miRNAs sorting and promotes HCC progression.

Keywords:  Exosome; Hepatocellular carcinoma; ST6Gal-I; Sialylation; miRNAs

DOI:  https://doi.org/10.1007/s13105-022-00917-1
Cell Biol Toxicol. 2022 Aug 17.

miR-222-3p-containing macrophage-derived extracellular vesicles confer gemcitabine resistance via TSC1-mediated mTOR/AKT/PI3K pathway in pancreatic cancer.

Yao Guo, Heshui Wu, Jiongxin Xiong, Shanmiao Gou, Jing Cui, Tao Peng.

  Gemcitabine resistance limits the efficacy of chemotherapy and maintains a challenge for treatment outcomes. Therefore, we aimed to clarify the downstream mechanisms underlying the role of miR-222-3p delivered by M2 macrophage-derived extracellular vesicles (M2 MDEs) in the chemoresistance of pancreatic cancer (PCa). We separated the mouse macrophages and polarized them to M2 phenotypes, from which the EVs were derived. miR-222-3p was highly expressed in M2 MDEs. M2 MDEs were internalized by PCa cells. miR-222-3p overexpressing M2 MDEs were treated with gemcitabine and co-cultured with PCa cells for in vitro experiments. Co-culture with M2 MDEs enriched with miR-222-3p suppressed the sensitivity to gemcitabine, accompanied by diminished apoptosis and promoted proliferation. Furthermore, the M2 MDEs and PCa cells were injected to mice with gemcitabine exposure for in vivo substantiation. The delivery of miR-222-3p inhibitor by M2 MDEs suppressed tumor growth and elevated sensitivity of cancer cells to gemcitabine. Moreover, miR-222-3p was indicated to target and suppress TSC1 expression, while miR-222-3p activated the PI3K/AKT/mTOR pathway. Together, miR-222-3p-containing M2 MDEs enhance chemoresistance in PCa through TSC1 inhibition and activation of the PI3K/AKT/mTOR pathway.

Keywords:  Drug resistance; Extracellular vesicles; Gemcitabine; M2 macrophages; MicroRNA-222-3p; Pancreatic cancer; TSC1

DOI:  https://doi.org/10.1007/s10565-022-09736-y
Theranostics. 2022 ;12(13): 5836-5855

Extracellular Vesicles and Circulating Tumour Cells - complementary liquid biopsies or standalone concepts?

Artur Słomka, Bingduo Wang, Tudor Mocan, Adelina Horhat, Arnulf G Willms, Ingo G H Schmidt-Wolf, Christian P Strassburg, Maria A Gonzalez-Carmona, Veronika Lukacs-Kornek, Miroslaw T Kornek.

  Liquid biopsies do promise a lot, but are they keeping it? In the past decade, additional novel biomarkers qualified to be called like that, of which, some took necessary hurdles resulting in FDA approval and clinical use. Some others are since a while around, well known and were once regarded to be a game changer in cancer diagnosis or cancer screening. But, during their clinical use limitations were observed from statistical significance and questions raised regarding their robustness, that eventually led to be dropped from associated clinical guidelines for certain applications including cancer diagnosis. The purpose of this review isn't to give a broad overview of all current liquid biopsy as biomarkers, weight them and promise a brighter future in cancer prevention, but rather to take a deeper look on two of those who do qualify to be called liquid biopsies now or then. These two are probably of greatest interest conceptually and methodically, and likely have the highest chances to be in clinical use soon, with a portfolio extension over their original conceptual usage. We aim to dig deeper beyond cancer diagnosis or cancer screening. Actually, we aim to review in depth extracellular vesicles (EVs) and compare with circulating tumour cells (CTCs). The latter methodology is partially FDA approved and in clinical use. We will lay out similarities as taking advantage of surface antigens on EVs and CTCs in case of characterization and quantification. But drawing readers' attention to downstream application based on capture/isolation methodology and simply on their overall nature, here apparently being living material eventually recoverable as CTCs are vs. dead material with transient effects on recipient cell as in case of EVs. All this we try to bring in perspective, compare and conclude towards which future direction we are aiming for, or should aim for. Do we announce a winner between CTCs vs EVs? No, but we provide good reasons to intensify research on them.

Keywords:  CTC; biomarker; ectosomes; exosomes; extracellular vesicles; liquid biopsy; microvesicles; personalized medicine

DOI:  https://doi.org/10.7150/thno.73400
J Exp Clin Cancer Res. 2022 Aug 19. 41(1): 253

Tumor associated macrophages-derived exosomes facilitate hepatocellular carcinoma malignance by transferring lncMMPA to tumor cells and activating glycolysis pathway.

Minghao Xu, Chenhao Zhou, Jialei Weng, Zhaoshuo Chen, Qiang Zhou, Jian Gao, Guoming Shi, Aiwu Ke, Ning Ren, Huichuan Sun, Yinghao Shen.

   BACKGROUND: Tumor-associated macrophages (TAMs), which form a large part of the tumor microenvironment, are normally regulated by metabolic reprogramming. However, the potential mechanisms of the immune-metabolism interaction between hepatocellular carcinoma (HCC) cells and TAMs remain unclear.
METHODS: The candidate long non-coding RNAs (lncRNAs) were screened by Smart-seq based scRNA-seq method and then validated by qPCR. Immunostaining analysis was done to examine the levels of markers for TAMs and glycolysis. Exosomes from primary TAMs of human HCC tissues were isolated by centrifugation, and their internalization with lncRNAs was confirmed by immunofluorescence. The underlying mechanism of TAMs-derived exosomal lncRNA to HCC was confirmed by luciferase reporter assay and RNA immunoprecipitation. Metabolism regulation was evaluated through glucose consumption, lactate productions and extracellular acidification rates (ECARs). Mouse xenograft models were used to elucidate the in vivo effect of candidate lncRNAs on tumor growth.
RESULTS: TAMs augment the aerobic glycolysis in HCC cells and their proliferation by the extracellular exosome transmission of a myeloid-derived lncRNA, M2 macrophage polarization associated lncRNA (lncMMPA). Mechanistically, lncMMPA not only could polarize M2 macrophage, but also could act as an microRNA sponge to interact with miR-548 s and increase the mRNA level of ALDH1A3, then further promote glucose metabolism and cell proliferation in HCC. Moreover, lncMMPA increased HCC cell multiplication through interacting with miR-548 s in vivo. Clinically, lncMMPA expression associates with glycolysis in TAMs and reduced survival of HCC patients.
CONCLUSION: LncMMPA plays an important role in regulating HCC malignancy and metabolic reprogramming of miR-548 s/ALDH1A3 pathway.

Keywords:  Hepatocellular carcinoma; LncMMPA; Metabolic reprogramming; Tumor associated macrophage

DOI:  https://doi.org/10.1186/s13046-022-02458-3
Clin Transl Oncol. 2022 Aug 18.

Cancer cell-derived exosomal LINC00313 induces M2 macrophage differentiation in non-small cell lung cancer.

Wencui Kong, Lei Zhang, Ying Chen, Zongyang Yu, Zhongquan Zhao.

   PURPOSE: Non-small cell lung cancer (NSCLC) is the major subtype of lung cancer, which is the leading cause of cancer death worldwide. Tumor-associated macrophages (TAMs) are one of the main non-tumor cells in the tumor microenvironment. Here, we investigated the effect of cancer cell-derived exosomal LINC00313 on the M2 macrophage differentiation in NSCLC and clarified its underlying mechanism.
METHODS: Flow cytometry, Western blotting, ELISA and immunohistochemical staining were performed to identify the macrophage phenotype by detecting the expression of M2 markers. The expression levels of LINC00313 and miR-135a-3p were measured by qRT-PCR, and luciferase reporter assay was used to validate the binding of lncRNA to miRNA, and miRNA to the target gene STAT6. The mouse-xenograft models were established by subcutaneous injection of the NCl-H1299 cells with stable overexpression or knockdown of LINC00313. GW4869 was injected intra-tumorally after tumor implantation.
RESULTS: It was found that the cancer cells promoted M2 macrophage differentiation by secreting exosomes. LINC00313 was overexpressed in H1299-derived exosomes, and its knockdown abolished the effect of H1299-induced M2 macrophage differentiation. LINC00313 sponged miR-135a-3p to increase the STAT6 expression, resulting in the M2 macrophage differentiation. LINC00313 promoted tumor progression and promoted the expression of M2 markers in isolated tumor macrophages. A novel regulatory mechanism of M2 macrophage differentiation in NSCLC was revealed. It was found that cancer cell-derived exosomal LINC00313 promoted M2 macrophage differentiation in NSCLC by up-regulating STAT6 as miR-135a-3p sponge.
CONCLUSIONS: This study provides a new mechanism and direction to prevent the M2 macrophage differentiation in NSCLC.

Keywords:  Exosome; LncRNA; Macrophage differentiation; Non-small cell lung cancer; microRNA

DOI:  https://doi.org/10.1007/s12094-022-02907-7
Int J Biol Sci. 2022 ;18(13): 5123-5135

Decrease of circARID1A retards glioblastoma invasion by modulating miR-370-3p/ TGFBR2 pathway.

Baisheng Li, Jiansheng Chen, Yi Wu, Honghai Luo, Yiquan Ke.

  Increasing evidence suggests that circular RNAs (circRNAs) are involved in regulating tumor biological activity. Glioblastoma (GBM) is one of the most lethal diseases characterized by highly aggressive proliferative and invasive behaviors. We aimed to explore how circRNAs influenced GBM biological activity. By circRNA array analysis we found that circARID1A was significantly up-regulated in GBM. Next, we found that circARID1A was up-regulated in GBM tissues and cell lines. Interfering with circARID1A inhibited the migration and invasion of a human GBM cell line U87. By performing dual-luciferase reporter assays, RNA pull-down and fluorescent in situ hybridization (FISH), we determined that circARID1A directly bound to miR-370-3p. Moreover, we confirmed that transforming growth factor beta receptor 2 (TGFBR2) was the target gene of miR-370-3p by performing RNA pull-down, dual-luciferase reporter assays and western blotting. Further experiments verified that circARID1A promoted GBM cell migration and invasion by modulating miR-370-3p/ TGFBR2 pathway. In addition, we demonstrated that silencing circARID1A restrain the growth of GBM in vivo. Finally, we showed that circARID1A was abundant in GBM cell derived exosomes. In conclusion, circARID1A participated in regulating migration and invasion of GBM via modulation of miR-370-3p/ TGFBR2 and thus may be a potential serum biomarker of GBM.

Keywords:  TGFBR2; circARID1A; exosomes; glioblastoma; miR-370-3p

DOI:  https://doi.org/10.7150/ijbs.66673
Cell Death Dis. 2022 Aug 19. 13(8): 725

Exosomal miR-155-5p derived from glioma stem-like cells promotes mesenchymal transition via targeting ACOT12.

Zixu Bao, Ning Zhang, Wanxiang Niu, Maolin Mu, Xiaoming Zhang, Shanshan Hu, Chaoshi Niu.

Tumor-associated exosomes play essential roles in intercellular communication and the foundation of cancer microenvironment in glioma. Many mRNAs, microRNAs (miRNAs) and proteins contained in tumor-associated exosomes can be transferred to recipient cells and contribute to the progression of tumor. Nevertheless, the cellular communication between malignant cells with different heterogeneities or characteristics and resultant tumor progression are still unclear in glioma. Here, we show that exosomes released from glioma stem-like cells (GSCs) contain a significant increasing level of miR-155-5p and could be horizontally transferred to surrounding glioma cells. High expression of miR-155-5p in plasma exosomes from patients was associated with glioma diagnosis and grading. Mechanically, we found that miR-155-5p markedly reduced the expression of acetyl-CoA thioesterase 12 (ACOT12), which played as a tumor suppressor in glioma. Furthermore, mesenchymal transition was significantly promoted in glioma cells treated with GSCs-derived exosomes. In conclusion, GSCs-derived exosomal miR-155-5p play a critical role in glioma progression and facilitating tumor aggressive growth by targeting ACOT12 and promoting mesenchymal transition. Exosomal miR-155-5p is also a potential predictive biomarker for glioma, which may provoke the development of novel diagnostic and therapeutic strategies against glioma.

DOI: https://doi.org/10.1038/s41419-022-05097-w
Can J Urol. 2022 Aug;29(4): 11224-11230

How I Use It: The Exosome Diagnostics (EPI) prostate cancer biomarker utility in urology and primary care.

Judd W Moul, Grannum R Sant.

Prostate-specific antigen (PSA) screening remains the mainstay for early detection of prostate cancer. Although PSA is a nonspecific prostate cancer biomarker, its specificity for high grade prostate cancer can be enhanced by pre-biopsy liquid biomarkers including the Exosome Dx Prostate IntelliScore (EPI) test. EPI is a stand-alone urine genomic test that measures 3 exosome-derived gene expression signatures without the need for digital rectal examination (DRE) or inclusion of standard of care parameters in the test algorithm. EPI has broad clinical utility as a risk stratification tool for clinically significant high grade prostate cancer in men considering diagnostic prostate biopsy (MRI-targeted and systematic biopsy). During the COVID-19 pandemic, the EPI At-Home Collection Kit was introduced and quickly became an important component of tele-urology. The EPI test has emerged as a prioritization tool for primary care referral to urologists and for prostate biopsy scheduling. EPI provides an objective and actionable genomic risk assessment tool for high grade prostate cancer and is a critical part of the informed decision-making regarding biopsy (targeted, systematic or both) in both urology and primary care practices.

Keywords: ExoDx (EPI); MRI; biomarkers; primary care; prostate cancer; prostate-specific antigen (PSA) screening; urology
J Exp Clin Cancer Res. 2022 Aug 20. 41(1): 254

Epstein-Barr viral product-containing exosomes promote fibrosis and nasopharyngeal carcinoma progression through activation of YAP1/FAPα signaling in fibroblasts.

Po-Ju Lee, Yun-Hua Sui, Tzu-Tung Liu, Ngan-Ming Tsang, Chen-Han Huang, Ting-Yi Lin, Kai-Ping Chang, Shu-Chen Liu.

   BACKGROUND: The progression of nasopharyngeal carcinoma (NPC) is profoundly affected by Epstein-Barr virus (EBV) infection. However, the role of EBV in the intercommunication between NPC and surrounding stromal cells has yet to be explored.
METHODS: NPC biopsies were obtained for immunohistochemical (IHC) analyses. Clinical correlations between the expression of active YAP1/FAPα and the fibrotic response and between YAP1/FAPα and the density of cytotoxic CD8a+ T lymphocytes were determined. Survival times based on IHC scores were compared between groups using Kaplan-Meier survival and log-rank tests. Independent prognostic factors for metastasis/recurrence-free survival and overall survival were identified using univariate and multivariate Cox regression models. Fibroblasts were isolated from human nasopharyngeal biopsies. Exosomes were purified from culture supernatants of EBV+-positive NPC cells. The effects of EBV product-containing exosomes on fibroblast activation, fibrotic response, tumor growth, immune response, and correlations between the expression of featured genes were investigated using gel contraction assays, ELISAs, EdU incorporation assays, real-time impedance assays, RNA sequencing, immunostaining, 3D cancer spheroid coculture systems, and an NPC xenograft model.
RESULTS: NPC patients who developed metastasis had significantly higher levels of active YAP1 and FAPα in their tumor stroma, which was further correlated with tumor fibrosis and poorer metastasis-free survival. Exosomes released from EBV+-NPC cells contained abundant FAPα protein and EBV-encoded latent membrane protein 1. Viral product-containing exosomes markedly enhanced the fibrotic response and tumor growth in a mouse xenograft model. IHC analyses of human NPC and NPC xenografts revealed positive correlations between levels of active YAP1 and FAPα, YAP1 and the fibrotic response, and FAPα and the fibrotic response. Mechanistic studies showed that treatment of fibroblasts with viral product-containing exosomes promoted the characteristics of cancer-associated fibroblasts by stimulating YAP1 signaling and the production of the immunosuppressive cytokines IL8, CCL2, and IL6. Inhibition of YAP1 activation markedly reversed these exosome-mediated protumoral effects, resulting in reduced contractility, inactivation of YAP1 signaling, and decreased production of immunosuppressive cytokines in fibroblasts. Furthermore, fibroblasts stimulated with these viral product-containing exosomes promoted NPC resistance to T cell-mediated cytotoxicity within tumor spheroids. In NPC tissues, a significant negative correlation was found between YAP1/FAPα and the density of CD8a+ T lymphocytes with a granzyme B signature.
CONCLUSION: EBV orchestrates interactions with the host and surrounding stroma by stimulating the functions of YAP1 and FAPα in fibroblasts through exosome cargos to create a more immunosuppressive, proinvasive microenvironment.

Keywords:  EBV; Epstein-Barr virus; Exosome; FAPα; Fibroblast; Fibroblast activation protein alpha; Fibrosis; NPC; Nasopharyngeal carcinoma; YAP1

DOI:  https://doi.org/10.1186/s13046-022-02456-5
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2022 Aug;30(4): 1188-1192

[Release of Exosomes Derived from Leukocyte-Depleted Red Cell Suspension and Its Regulation on Hematological Tumor Cells].

Hao-Bo Huang, Li-Ping Fan, Qiu-Yan Lin, Hui-Wen Huang, Dan-Hui Fu.

   OBJECTIVE: To investigate the release of exosome (Exo) from leukocyte-depleted red cell suspension (LDRCS) at different storage time and its regulation on proliferation of hematological tumor cells and possible mechanism.
METHODS: The Exo (RBC-Exo) in LDRCS at different storage time was obtained by ultracentrifugation, and the morphology and immunological marker of RBC-Exo were detected by transmission electron microscopy and Western blot, respectively. The particle size distribution of RBC-Exo in LDRCS at different storage time was detected by Dynamic Light Scattering. CCK-8 assay was used to explore the effect of RBC-Exo on hematological tumor cell proliferation. Western blot was used to detect the expression of proliferation-related proteins in hematological tumor cells after co-culture with RBC-Exo.
RESULTS: RBC-Exo was isolated, which was characterized by cup-like shape, particle size distribution ranged from 20 to 200 nm, CD63/TSG101 enriched, Calnexin negative, CD235a positive and CD41 negative. The particle size distribution of RBC-Exo from LDRCS between middle was not significantly different and late stored stage. But the particle size distribution of RBC-Exo at middle-late stored stage(>14 d) was larger than that at early stored stage (≤14 days). Compared with the control group, RBC-Exo could significantly promote the proliferation of HBL1, U2932 and Jurkat cells. Compared with the control group, the cycle-related protein P21 was significantly down-regulated in HBL1, U2932 and Jurkat cells after co-culture with RBC-Exo for 3 days, while the anti-apoptotic protein BCL-2 was not changed significantly.
CONCLUSION: The morphology of RBC-Exo from LDRCS at middle-late stored stage was different from that at early stored stage. RBC-Exo could promote the proliferation of hematological tumor cells, possibly by regulating the expression of cycle-associated protein P21.

Keywords:  exosome ; leukocyte-depleted red cell suspension ; proliferation ; tumor cell

DOI:  https://doi.org/10.19746/j.cnki.issn.1009-2137.2022.04.033