Abstract
Transmembrane integrin receptors mediate cell–extracellular matrix as well as cell–cell adhesion. As placental trophoblast cells undergo differentiation they display changes in integrin expression or switching, but the mechanism(s) of integrin activation that supports this differentiation is still unknown. The Fermitin family of adapter proteins (FERMT 1–3) are integrin activators that mediate integrin-mediated signaling. In this study, we examined the spatiotemporal pattern of expression of FERMT1 in human chorionic villi throughout gestation and its role in HTR8-SVneo substrate adhesion and invasion. Placental villous tissue was obtained from patients undergoing elective terminations at weeks 8–14, as well as from term deliveries at weeks 37–40 and analyzed by immunofluorescence. Additionally, HTR8-SVneo trophoblast cells were transfected with FERMT1-specific siRNA or non-targeting siRNA (control) and used in cell-substrate adhesion as well as invasion assays. FERMT1 was primarily localized to membrane-associated regions at the base or around the periphery of the villous cytotrophoblast and proximal as well as distal cell column trophoblast. FERMT1 was also localized to endothelial cells of blood vessels in chorionic villi. siRNA-mediated depletion of FERMT1 in HTR8-SVneo cells did not markedly alter HTR8-SVneo cell-substrate adhesion but did significantly decrease invasion (P < 0.05) compared to control cells. These novel findings identify the presence of the integrin activator FERMT1 in trophoblast cells and that FERMT1 can regulate HTR8-SVneo cell invasion. FERMT1 may directly influence integrin activation and the subsequent integrin-mediated signaling and differentiation that underlies the acquisition of the invasive trophoblast phenotype in vivo.
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Funding
Funded by the Canadian Institutes of Health Research (Grant # 101051), Saskatchewan Health Research Foundation (Grant # 2776), the Western College of Veterinary Medicine Research Trust (Grant # 417785), and the Canada Foundation for Innovation John R. Evans Leaders Fund (Project # 32512) to DJM.
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EK and DJM designed experiments and EK, GBH, EIM, and DJM performed experiments. EK and DJM interpreted results and wrote the manuscript. EK, EIM and DJM critically reviewed the manuscript.
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418_2021_1977_MOESM1_ESM.tiff
Supplementary file1 Supplementary Fig 1 Comparison of HLA-G detection in human placental tissue with two different antisera. Representative images at week (W) 9 (a–c) and W13 (d–f) of gestation are shown. Cell column trophoblast (a–c) and extravillous trophoblast (d–f) were incubated with a rabbit polyclonal anti-human leukocyte antigen-G (HLA-G)-specific antiserum (see Table 1; green) and a mouse monoclonal (clone 4H84) anti-HLA-G specific antiserum (MABF2169, Sigma Aldrich, red). HLA-G was detectable in the same distal cell column trophoblast (dCCT) and extravillous trophoblast cells with the two antisera. HLA-G was not detectable in proximal CCT (pCCT). IgG: mouse and rabbit immunoglobulins used in place of primary antisera. Nuclei were stained with DAPI where indicated. Scale bar = 50 μm (TIFF 4895 KB)
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Kawamura, E., Hamilton, G.B., Miskiewicz, E.I. et al. Examination of FERMT1 expression in placental chorionic villi and its role in HTR8-SVneo cell invasion. Histochem Cell Biol 155, 669–681 (2021). https://doi.org/10.1007/s00418-021-01977-y
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DOI: https://doi.org/10.1007/s00418-021-01977-y