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Circulating extracellular vesicle protein biomarker associated with early cancer lesions for early detection of high-grade serous ovarian cancer

ABSTRACT: Small extracellular vesicles (sEVs), lipid-bilayer delimited particles (50-200 nm) released by cells, are considered a new class of liquid biopsy biomarkers for stealth cancers, e.g., HGSOC. HGSOC originates from the fallopian tube (FT) and progresses from p53 signatures to a precursor lesion called serous tubal intraepithelial carcinoma (STIC). We hypothesize that sEVs play a role in ovarian pathogenesis, carry cargo reflective of the site of origin, and are biomarkers for early detection. To advance this idea, we established a case-control study cohort of archival plasma samples from 30 HGSOC patients (10 early-stage and 20 late-stage) and 40 healthy controls. sEVs were isolated using size exclusion chromatography, and proteomic profiles were established using mass spectrometry (LC-MS/MS). 1,078 EV proteins (exo-proteins) were identified across all samples, with 52 upregulated exo-proteins in early-stage HGSOC, and 59 upregulated exo-proteins in late-stage HGSOC (logFC>1, p<0.05). Upregulated exo-proteins were prioritized based on FT origin and tissue expression in STIC lesions. Seven candidate exo-proteins were validated by immunohistochemistry in FT with STIC lesions and HGSOC tissues and by western blot in FT/HGSOC cell-derived EVs. In summary, our EV proteomics using early-stage HGSOC clinical samples uncovered exo-biomarkers for early detection; potentially while still confined to the FT.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Blood Plasma

SUBMITTER: Dennis Province

LAB HEAD: Andrew K. Godwin

PROVIDER: PXD067228 | Pride | 2026-01-08

REPOSITORIES: Pride

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Source:

			Action	DRS
	DDA_EV_RAW_MQ_Phospho_STY_Sites.txt	Txt
	DDA_EV_RAW_MQ_proteinGroups.txt	Txt
	DDA_tissue_RAW_MQ_Phospho_STY_Sites.txt	Txt
	DDA_tissue_RAW_MQ_proteinGroups.txt	Txt
	GODWIN_022123_TISSUE_DDA_SF01.mgf	Mgf

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Project description:This project aimed at identifying developmental stage specific transcript profiles for catecholaminergic neurons in embryos and early larvae of zebrafish (Danio rerio). Catecholaminergic neurons were labeled using transgenic zebrafish strains to drive expression of GFP. At stages 24, 36, 72 and 96 hrs post fertilization, embryos were dissociated and GFP expressing cells sorted by FACS. Isolated RNAs were processed using either polyA selection and libray generation or NanoCAGE. This is the first effort to determine stage specific mRNA profiles of catecholaminergic neurons in zebrafish. Catecholaminergic neurons were labeled by four different strategies: (1) 24 hrs old embryos: we used the ETvmat2:GFP transgenic line (Wen et al. 2007). Visualization of monoaminergic neurons and neurotoxicity of MPTP in live transgenic zebrafish. Dev Biol. 2008 Vol 314 p84-92) which at this early stage labels catecholaminergic neurons in posterior tuberculum and locus coeruleus; (2) 24 hrs old embryos: we used Tg(otpb.A:egfp)zc48 transgenic line (Fujimoto et al. Identification of a dopaminergic enhancer indicates complexity in vertebrate dopamine neuron phenotype specification. Dev Biol 2011, Vol 352, p393–404) which at this stage label ventral diencephalic dopaminergic neurons and some preoptic neurons. (3) For 72 and 96 hrs old zebrafish larvae we used a th:GFP BAC transgenic lines that labels catecholaminergic neurons (Tay et al., Comprehensive catecholaminergic projectome analysis reveals single-neuron integration of zebrafish ascending and descending dopaminergic systems. Nat Comms 2011 Vol 2, 171; also: T. Leng and W. Driever, unpublished). (4) for the 36 and 48 hrs old zebrafish larvae we used a th:Gal4VP16 driver and UAS:EGFP responder transgenic line system to label catecholaminergic cells (Fernandes et al., Deep brain photoreceptors control light-seeking behavior in zebrafish larvae. Curr Biol. 2012 Vol 22 DOI 10.1016/j.cub.2012.08.016). We used the different transgenic lines, because lines (3) and (4) do not efficiently label catecholaminergic neurons at early stages, while lines (1) and (2) also have GFP expression in several other non-catecholaminergic populations at later stages of development. Embryos were dissociated and catecholaminergic neurons were FACS sorted from GFP-tagged zebrafish (Manoli and Driever, 2012, Cold Spring Harbor Protoc. DOI 10.1101/pdb.prot069633). RNA was either processed for NanoCAGE, or mRNA was isolated and amplified. cDNA was sequenced by Illumina technique. This data submission is a series of data files consisting of three independent experiments with diffrent RNA-Seq depth: Samples 1-4 (NanoCage): Samples 5-8 (RNA-Seq high read numbers), and SAmples 9-12 (RNA-Seq low read numbers).

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Circulating extracellular vesicle protein biomarker associated with early cancer lesions for early detection of high-grade serous ovarian cancer

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