Proteomic Profiling of Fallopian Tube-Derived Extracellular Vesicles Using a Microfluidic Tissue-on-Chip System

Abstract

The human fallopian tube epithelium (hFTE) is the site of fertilization, early embryo development, and the origin of most high-grade serous ovarian cancers (HGSOCs). Little is known about the content and functions of hFTE-derived small extracellular vesicles (sEVs) due to the limitations of biomaterials and proper culture methods. We have established a microfluidic platform to culture hFTE for EV collection with adequate yield for mass spectrometry-based proteomic profiling, and reported 295 common hFTE sEV proteins for the first time. These proteins are associated with exocytosis, neutrophil degranulation, and wound healing, and some are crucial for fertilization processes. In addition, by correlating sEV protein profiles with hFTE tissue transcripts characterized using GeoMx® Cancer Transcriptome Atlas, spatial transcriptomics analysis revealed cell-type-specific transcripts of hFTE that encode sEVs proteins, among which, FLNA, TUBB, JUP, and FLNC were differentially expressed in secretory cells, the precursor cells for HGSOC. Our study provides insights into the establishment of the baseline proteomic profile of sEVs derived from hFTE tissue, and its correlation with hFTE lineage-specific transcripts, which can be used to evaluate whether the fallopian tube shifts its sEV cargo during ovarian cancer carcinogenesis and the role of sEV proteins in fallopian tube reproductive functions.

Oral exposure to bisphenol S is associated with alterations in the oviduct proteome of an ovine model, with aggravated effects in overfed females

“…2 and Table S1). In addition, 15% of proteins (229/1563) were previously reported in oviduct extracellular vesicles (EVs) from the bovine [ 31 ] or human [ 32 ] species (Fig. 2 and Table S1).…”

Oral exposure to bisphenol S is associated with alterations in the oviduct proteome of an ovine model, with aggravated effects in overfed females

“…Furthermore, 7% of identi ed proteins (103/1,563) contained a signal peptide and were predicted to be classically secreted, while 29% of proteins (453/1,563) were predicted to be secreted by unconventional pathways in the oviduct uid (Fig 2 and Table S1). In addition, 15% of proteins (229/1,563) were previously reported in oviduct extracellular vesicles (EVs) from the bovine [30] or human [31] species (Fig 2 and Table S1).…”

“…Brie y, FASTA sequences of all identi ed proteins were retrieved from Uniprot ID mapping online tool, and were used as inputs into the Outcyte online tool (version 1.0; http://www.outcyte.com/) [54] and SignalP in Eukarya organism (version 6.0; https://services.healthtech.dtu.dk/services/SignalP-6.0/) [55]. Prediction of secretion by oviduct EVs was performed by comparing the list of genes of all identi ed proteins with the ones previously described in bovine [30] and human [31] oviduct EVs.…”

“…The number of proteins (with a signal peptide) and unconventionally secreted were predicted using Outcyte and Signal P online tools. The number of proteins identi ed in oviduct extracellular vesicles were assessed using previous proteomic data in bovine [30] and human [31] oviduct extracellular vesicles.…”

Opportunities involving microfluidics and 3D culture systems to the in vitro embryo production

“…By creating a more biomimetic environment with precise control over culture ( Wale and Gardner, 2016 ), the combined approach has the potential to improve embryo development and quality. The integration of these technologies was already used to create endometrial ( Ahn et al, 2021 ; De Bem et al, 2021 ; Xiao et al, 2017 ) and oviductal ( Ferraz et al, 2020b ; Ferraz et al, 2018b , 2017 ; Xiao et al, 2017 ; Zha et al, 2023 ) models which can provide valuable insights into embryo-maternal communication and also to recapitulate follicle growth and ovulation in vitro ( Laronda et al, 2017 ; Xiao et al, 2017 ) ( Figure 3 ). However, several challenges remain, including technical and engineering complexities, standardization and reproducibility issues, and the need for validation against conventional IVEP methods.…”