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Details	EV-TRACK ID	Experiment nr.	Species	Sample type	Separation protocol	First author	Year	EV-METRIC
	EV170041	1/1	Homo sapiens	'SW480 Dukes' type B, colorectal adenocarcinoma;SW620, colon; derived from metastatic site: lymph node	DG	Yingkuan Shao	2018	87%
Study summary Full title Colorectal cancer-derived small extracellular vesicles establish an inflammatory premetastatic niche in liver metastasis All authors Yingkuan Shao, Ting Chen, Xi Zheng, Sheng Yang, Kailun Xu, Xuewen Chen, Fei Xu, Lantian Wang, Yanwei Shen, Tingyang Wang, Mengwen Zhang, Wangxiong Hu, Chenyang Ye, XiaoFang Yu, Jimin Shao, Shu Zheng Journal Carcinogenesis Abstract Liver metastases develop in more than half of the patients with colorectal cancer (CRC) and are asso (show more...)Liver metastases develop in more than half of the patients with colorectal cancer (CRC) and are associated with a poor prognosis. The factors influencing liver metastasis of CRC are poorly characterized, but this information is urgently needed. We have now discovered that small extracellular vesicles (sEVs; exosomes) derived from CRC can be specifically targeted to liver tissue and induce liver macrophage polarization toward an interleukin-6 (IL-6)-secreting proinflammatory phenotype. More importantly, we found that microRNA-21-5p (miR-21) was highly enriched in CRC-derived sEVs and was essential for creating a liver proinflammatory phenotype and liver metastasis of CRC. Silencing either miR-21 in CRC-sEVs or Toll-like receptor 7 (TLR7) in macrophages, to which miR-21 binds, abolished CRC-sEVs' induction of proinflammatory macrophages. Furthermore, miR-21 expression in plasma-derived sEVs was positively correlated with liver metastasis in CRC patients. Collectively, our data demonstrate a pivotal role of CRC-sEVs in promoting liver metastasis by inducing an inflammatory premetastatic niche through the miR-21-TLR7-IL-6 axis. Thus, sEVs-miR-21 represents a potential prognostic marker and therapeutic target for CRC patients with liver metastasis. (hide) EV-METRIC 87% (98th percentile of all experiments on the same sample type) Reported Not reported Not applicable EV-enriched proteins Protein analysis: analysis of three or more EV-enriched proteins non EV-enriched protein Protein analysis: assessment of a non-EV-enriched protein qualitative and quantitative analysis Particle analysis: implementation of both qualitative and quantitative methods. For the quantitative method, the reporting of measured EV concentration is expected. electron microscopy images Particle analysis: inclusion of a widefield and close-up electron microscopy image density gradient Separation method: density gradient, at least as validation of results attributed to EVs EV density Separation method: reporting of obtained EV density ultracentrifugation specifics Separation method: reporting of g-forces, duration and rotor type of ultracentrifugation steps antibody specifics Protein analysis: antibody clone/reference number and dilution lysate preparation Protein analysis: lysis buffer composition Study data Sample type Cell culture supernatant Sample origin Control condition Focus vesicles small extracellular vesicles Separation protocol Separation protocol Gives a short, non-chronological overview of the different steps of the separation protocol. dUC = (Differential) (ultra)centrifugation DG = density gradient UF = ultrafiltration SEC = size-exclusion chromatography IAF = immuno-affinity capture DG Protein markers EV: Alix/ HSP70/ CD63/ CD9 non-EV: Calreticulin Proteomics no Show all info Study aim Function, Biomarker, Mechanism of uptake/transfer, Identification of content (omics approaches) Sample Species Homo sapiens Sample Type Cell culture supernatant EV-producing cells 'SW480 Dukes' type B, colorectal adenocarcinoma / SW620, colon / derived from metastatic site: lymph node EV-harvesting Medium EV-depleted serum Preparation of EDS overnight (16h) at >=100,000g Cell viability (%) NA Separation Method Density gradient Only used for validation of main results Yes Density medium 69.83 Type Discontinuous Number of initial discontinuous layers 3 Lowest density fraction 0.25M Highest density fraction 1.3M Sample volume (mL) 1 Orientation Bottom-up (sample migrates upwards) Rotor type SW 41 Ti Speed (g) 100000 Duration (min) 150 Fraction volume (mL) 2 Fraction processing Centrifugation Pelleting: volume per fraction 5 Pelleting: duration (min) 150 Pelleting: rotor type MLS-50 Pelleting: speed (g) 100000 Pelleting: adjusted k-factor 142.1 Pelleting-wash: volume per pellet (mL) 1 Pelleting-wash: duration (min) 150 Pelleting-wash: rotor type 69.83 Pelleting-wash: speed (g) MLA-130 Pelleting-wash: adjusted k-factor 69.83 Characterization: Protein analysis Protein Concentration Method BCA Protein Yield (µg) 2-5 Western Blot Detected EV-associated proteins Alix, CD9, CD63, HSP70 Not detected contaminants Calreticulin Characterization: RNA analysis Database Yes Proteinase treatment Yes Moment of Proteinase treatment After Proteinase type Proteinase K Proteinase concentration 10 RNAse treatment Yes Moment of RNAse treatment After RNAse type RNase A RNAse concentration 10 Characterization: Lipid analysis No Characterization: Particle analysis NTA Report type Size range/distribution Reported size (nm) 30-160 EV concentration Yes Particle yield 1.0-2.0E11 EM EM-type Transmission-EM Image type Wide-field Report size (nm) 30-160 EV concentration Yes

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EV-TRACK ID	EV170041
species	Homo sapiens
sample type	Cell culture
cell type	'SW480 Dukes' type B colorectal adenocarcinoma SW620 colon derived from metastatic site: lymph node
condition	Control condition
separation protocol	DG
Exp. nr.	1
EV-METRIC %	87