Exploring the potential of exosomes in diagnosis and drug delivery for pancreatic ductal adenocarcinoma

Pancreatic cancer (PC) is a cancer of the digestive system, and pancreatic ductal adenocarcinoma (PDAC) accounts for approximately 90% of all PC cases. Exosomes derived from PDAC (PDAC-exosomes) promote PDAC development and metastasis. Exosomes are nanoscale vesicles secreted by most cells, which can carry biologically active molecules and mediate communication and cargo transportation among cells. Recent studies have focused on transforming exosomes into good drug delivery systems (DDSs) to improve the clinical treatment of PDAC. This review considers PDAC as the main research object to introduce the role of PDAC-exosomes in PDAC development and metastasis. This review focuses on the following two themes: (a) the great potential of PDAC-exosomes as new diagnostic markers for PDAC, and (b) the transformation of exosomes into potential DDSs.     

外泌体在肿瘤细胞与TAMs之间相互作用中“桥梁”和调控作用的研究进展北大核心

外泌体是一类直径为30~100 nm的圆盘囊泡,其内包含许多组分,诸如复杂RNA和蛋白质等,主要参与细胞间的信号转导。肿瘤相关巨噬细胞(tumor-associated macrophages,TAMs)是肿瘤微环境中普遍存在的巨噬细胞,通过对肿瘤生长、免疫逃逸、侵袭和转移、耐药性等多方面的作用影响肿瘤进程。外泌体在肿瘤相关巨噬细胞的招募、极化及抗肿瘤免疫调控等方面发挥着重要的调节功能。同时,TAMs以外泌体为媒介作用于肿瘤细胞,从而构成了外泌体、TAMs与肿瘤细胞之间相互作用的调控通路。综上所述,本文旨在阐明肿瘤细胞与TAMs之间,以外泌体为“桥梁”相互影响的潜在机制,以及靶向肿瘤细胞和TAMs来源的外泌体在恶性肿瘤治疗中的展望。     

外泌体在卵巢癌诊疗及转移中的研究进展

卵巢癌是妇科肿瘤领域内的一个热点主题。该种恶性肿瘤疾病的发病率高且隐匿,缺乏有效的早期发现及时诊断并治疗的方式,病情容易加重发生远处转移。因此罹患卵巢癌的女性5年生存率比较低。外泌体属于一类典型的囊性泡样的小体,参与肿瘤的发病及转移机制且在肿瘤疾病的治疗中起着重要作用。本篇综述主要介绍癌细胞外泌体在卵巢恶性肿瘤疾病的诊断、转移及治疗中的研究现状。     

Prodrug suicide gene therapy for cancer targeted intracellular by mesenchymal stem cell exosomes

The natural behavior of mesenchymal stem cells (MSCs) and their exosomes in targeting tumors is a promising approach for curative therapy. Human tumor tropic mesenchymal stem cells (MSCs) isolated from various tissues and MSCs engineered to express the yeast cytosine deaminase::uracil phosphoribosyl transferase suicide fusion gene (yCD::UPRT-MSCs) released exosomes in conditional medium (CM). Exosomes from all tissue specific yCD::UPRT-MSCs contained mRNA of the suicide gene in the exosome's cargo. When the CM was applied to tumor cells, the exosomes were internalized by recipient tumor cells and in the presence of the prodrug 5-fluorocytosine (5-FC) effectively triggered dose-dependent tumor cell death by endocytosed exosomes via an intracellular conversion of the prodrug 5-FC to 5-fluorouracil. Exosomes were found to be responsible for the tumor inhibitory activity. The presence of microRNAs in exosomes produced from naive MSCs and from suicide gene transduced MSCs did not differ significantly. MicroRNAs from yCD::UPRT-MSCs were not associated with therapeutic effect. MSC suicide gene exosomes represent a new class of tumor cell targeting drug acting intracellular with curative potential.     

HepG2细胞外泌体冲击树突状细胞介导的抗肿瘤作用

目的：分析来源于肝癌HepG2细胞外泌体的免疫原性物质，为基于树突状细胞（DCs）的肝癌免疫治疗寻找合适的肿瘤抗原提供思路。方法：用透射电镜和Western blot方法鉴定肝癌HepG2细胞外泌体的形态和蛋白表达；用终浓度为8 μg/mL的外泌体冲击DCs，流式细胞术检测DCs表面分子的变化；将外泌体冲击的DCs与T淋巴细胞共培养5 d，采用羧基荧光素二乙酸盐琥珀酰亚胺酯（CFSE）荧光染色法检测淋巴细胞增殖，Annexin-V/PI双染法检测淋巴细胞对肿瘤的杀伤效应。结果：肝癌HepG2细胞的外泌体表达CD63、CD81标志性蛋白，同时携带大量的肿瘤抗原甲胎蛋白（AFP）和热休克蛋白HSP70、HSP90，不表达细胞蛋白calnexin。与未成熟DCs组比较，外泌体冲击DCs后上调其表面分子如CD83、CD80、CD86的表达（P < 0.01）；且可促进T淋巴细胞增殖（P < 0.01），增加T细胞介导的肿瘤特异性和非特异性杀伤效应（P < 0.01）。结论：肝癌HepG2细胞外泌体可能携带大量肿瘤抗原，刺激DCs成熟，可能是基于DCs的肝癌或其他肿瘤免疫治疗潜在的抗原谱。     

Exosomes secreted by prostate cancer cells under hypoxia promote matrix metalloproteinases activity at pre-metastatic niches

Approximately, 30 000 men die from prostate cancer (PCa) every year in the United States, mainly due to the metastasis. Thus, the key events associated with PCa metastasis are under rigorous investigation, with recent studies showing that preparation of pre-metastatic niches (PMN) in distant organs is an important step. However, the molecular basis for PMN preparation is still unclear. Hypoxia in primary tumors promotes aggressiveness; however, its precise role in metastasis is not clear. We recently reported that exosomes secreted by PCa cells under hypoxia promote stemness and invasiveness in naïve PCa cells; however, whether these extracellular vesicles also influence PMN remains unknown. In the present study, we isolated exosomes from human PCa PC3 cells under normoxic (21% O₂, exosomes secreted under normoxic condition [Exo^Normoxic]) and hypoxic (1% O₂, exosomes secreted under hypoxic condition [Exo^Hypoxic]) conditions, and characterized their effect (10 µg exosomes, intraperitoneal (IP) treatment every 48 hours for 4 weeks) on key biomarkers associated with PMN in nude mice. Whole animal fluorescence imaging showed that Exo^Hypoxic treatment promotes matrix metalloproteinases (MMPs) activity in several putative metastatic sites. Histological studies confirmed that Exo^Hypoxic treatment enhanced the level of MMP2, MMP9, and extracellular matrix proteins (fibronectin and collagen) as well as increased the number of CD11b+ cells at selective PMN sites. Furthermore, proteomic profiling of exosomes by liquid chromatography/mass spectrometry identified cargo proteins in Exo^Normoxic and Exo^Hypoxic as well as distinct canonical pathways targeted by them. These results suggest that exosomes secreted by PCa cells under hypoxia plausibly remodel distant PMN, and thus, could be a potential target to control metastatic PCa.     

A new player in the game: platelet-derived extracellular vesicles in dengue hemorrhagic fever

Abstract

Thrombocytopenia and vascular leakage are clinical hallmarks in dengue hemorrhagic fever. Sung et al. present a new mechanism where platelet-derived extracellular vesicles participate in increasing vascular permeability during dengue virus infection in mice.     

Urine and Serum Exosomes as Novel Biomarkers in Detection of Bladder Cancer

Background: The gold standard for initial clinical diagnosis of bladder cancer involves cystoscopic examination of bladder and histological evaluation of tissues. There is a critical need to identify non-invasive and sensitive biomarkers. Early detection is essential challenge in diagnosis and surveillance of bladder carcinoma. Exosomes are nano- sized vesicles present in many biological fluids and have significant role in cancer. Thus, quantification of exosomes in different stages of bladder cancer may be of critical concern for clinical diagnosis and prognosis. Methods: Tumor derived exosomes levels in urine and serum samples of 70 bladder cancer Egyptian patients from stages T0-T3 and 12 healthy control people were measured using ELISA technique. Results: When compared to health subjects, exosomes levels in bladder cancer patients were increased in urine and serum samples at different stages of the disease. A gradual increase in tumor derived exosomes in serum (1.21, 3.31, 4.71, 6.47µg/ml) and urine (1.59, 2.84, 4.75, 6.67µg/ml) was observed comparative to invasiveness of tumor (T0-T3). Serum was more specific (100%) sample for detection of exosomes in bladder cancer. Conclusion:  our findings suggest that tumor derived exosomes may offer a convenient tool for early diagnosis and monitoring of bladder cancer.     

Exosomes from miRNA‐126‐modified endothelial progenitor cells alleviate brain injury and promote functional recovery after stroke

AimsWe previously showed that the protective effects of endothelial progenitor cells (EPCs)‐released exosomes (EPC‐EXs) on endothelium in diabetes. However, whether EPC‐EXs are protective in diabetic ischemic stroke is unknown. Here, we investigated the effects of EPC‐EXs on diabetic stroke mice and tested whether miR‐126 enriched EPC‐EXs (EPC‐EXs^miR126) have enhanced efficacy.MethodsThe db/db mice subjected to ischemic stroke were intravenously administrated with EPC‐EXs 2 hours after ischemic stroke. The infarct volume, cerebral microvascular density (MVD), cerebral blood flow (CBF), neurological function, angiogenesis and neurogenesis, and levels of cleaved caspase‐3, miR‐126, and VEGFR2 were measured on day 2 and 14.ResultsWe found that (a) injected EPC‐EXs merged with brain endothelial cells, neurons, astrocytes, and microglia in the peri‐infarct area; (b) EPC‐EXs^miR126 were more effective than EPC‐EXs in decreasing infarct size and increasing CBF and MVD, and in promoting angiogenesis and neurogenesis as well as neurological functional recovery; (c) These effects were accompanied with downregulated cleaved caspase‐3 on day 2 and vascular endothelial growth factor receptor 2 (VEGFR2) upregulation till day 14.ConclusionOur results indicate that enrichment of miR126 enhanced the therapeutic efficacy of EPC‐EXs on diabetic ischemic stroke by attenuating acute injury and promoting neurological function recovery.