Targeting Underglycosylated MUC1 for the Selective Capture of Highly Metastatic Breast Cancer Cells Under Flow

Abstract

Abstract-The underglycosylated form of the MUC1 glycoprotein, uMUC1, has been identified as a ligand for both Eselectin and ICAM-1 and can play multiple potential roles during rolling and firm adhesion events in the metastatic cascade. Using flow cytometry and confocal microscopy, the T47D and ZR-75-1 cell lines were verified to highly express uMUC1, however it was found that only ZR-75-1 cells expressed the E-selectin binding moiety sialyl Lewis x (sLe x ). Furthermore, perfusing T47D cells through E-selectin coated microtubes resulted in fast rolling velocities and low numbers of interacting cells and blocking uMUC1 with the SM3 antibody had no effect. ZR-75-1 cells, on the other hand, were highly dependent on the E-selectin:uMUC1 interaction as exemplified by significant increases in cell rolling velocities and decreases in the number of interacting cells when blocking with SM3 or when uMUC1 expression was knocked down via siRNA transfection. Whereas uMUC1 interactions with E-selectin supported cell rolling, P-selectin: uMUC1 interactions exclusively facilitated cell tethering, while Lselectin surfaces supported no cell adhesive interactions. These experimental observations are consistent with molecular dynamics simulations of uMUC1 bound to E-, P-, and L-selectin where the degree of residue contact correlated with the differential adhesion of uMUC1 to each selectin. Finally, an E-selectin and SM3 combined surface coating captured approximately 30% of the total number of interacting cancer cells comparable to the number of adhered cells when utilizing E-selectin and ICAM-1 combined surfaces. The Eselectin/SM3 surface strategy offers a viable method to selectively capture cancer cells from whole blood samples.

A Comprehensive Investigation to Reveal the Relationship Between Plasmacytoid Dendritic Cells and Breast Cancer by Multiomics Data Analysis

“…L-selectin (SELL) was first discovered as a homing receptor on lymphocytes and later found to be expressed on various white blood cells. Chu et al documented that SELL immunodepletion inhibited MDA-MB-231 cell migration ( Laubli and Borsig, 2010 ; Geng et al, 2013 ; Chu et al, 2014 ). SPI1 transcription factors are closely related to developmental processes.…”

Galectin-1 Influences Breast Cancer Cell Adhesion to E-selectin Via Ligand Intermediaries

“…10): (i) Gal-1 binds to a shared ligand thereby causing a conformational change in the shared ligand and exposing the E-selectin preferred site to bind in monolayer adhesion; or (ii) Gal-1 binds to similar yet distinct ligand sets that would cause a signal cascade increase of separate E-selectin ligands within a domain to cluster or activate, thereby aiding in increased adhesion events. Possible ligand targets can be glycolipids, 31 MUC1, 22 or CD44/HCELL 32 however the exact effect of Gal-1 on these ligands would need to be identified.…”

“…6,[11][12][13]16,17,35 The distinctions between E-selectin and Gal-1 are varied, including general structure, functions within the system, and contributions to pro-inflammatory adhesion versus anti-inflammatory adhesion regulation of T cells, respectively. 13,15,22,23,26,37 While these two molecules have seemingly little in common, other than being lectins, there are key points that allude to a more dynamic relationship during breast cancer metastasis. Beyond their upregulation during cancer, especially more malignant cancers, 2,5,7,12,13,18,31,33,35 E-selectin and Gal-1 ligands seem to share the LacNAc backbone as a main carbohydrate skeletal feature.…”

<em>N</em>-(3-hydroxymethyl-&beta;-carboline-1-yl-ethyl-2-yl)-L-Phe: development toward a nanoscaled antitumor drug capable of treating complicated thrombosis and inflammation

“…According to the epidemiological and clinical estimation, ~25% of cancers are linked to chronic and acute inflammations 5–7. It has been well documented that P-selectin triggers the adhesion of leukocytes toward platelets, cancer cells, endothelial cells and other leukocytes at the thrombosis sites, inflammation tissues and tumor tissues 8–12. In addition, inhibiting P-selectin-mediated interactions of tumor cells with blood constituents results in attenuation of tumor metastasis 11,13–16.…”