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Series GSE76405 Query DataSets for GSE76405
Status Public on Dec 30, 2015
Title Generation of a monoclonal antibody recognizing the CEACAM glycan structure and inhibiting adhesion using cancer tissue-originated spheroid as an antigen
Organism Homo sapiens
Experiment type Expression profiling by array
Summary Spheroids cultured directly from tumours can better reflect in vivo tumour characteristics than two-dimensional monolayer culture or three-dimensional culture of established cell lines. In this study, we generated antibodies by directly immunizing mice with cancer tissue-originated spheroids from colorectal cancer. We performed phenotypic screening via recognition of the surface of the spheroids and inhibition of their adhesion to extracellular matrices to identify a monoclonal antibody, clone 5G2. The antibody inhibited cell migration in two-dimensional culture and promoted cell detachment. Western blotting and immunohistochemistry detected the 5G2 signal in many colorectal cancer spheroids, as well as patient tumours, but the signal was reduced in various cell lines. The expression pattern also changed when spheroids were cultured in two-dimensional conditions. We found that 5G2 recognized the N-glycan structure, including Lea and Lec, and their major carrier proteins were CEACAM5 and CEACAM6. Translocation of integrin 4 from the lateral membrane to the extracellular matrix contact interface in the spheroids was delayed by pre-incubation with 5G2. Thus, cancer tissue-originated spheroids can be a useful antigen for generating novel anti-cancer antibodies as the glycan structure is lost in cell lines but retained in cancer tissue-originated spheroids.
Overall design Gene expression was compared in cancer tissue-originated spheroids (CTOSs) under floating conditions, under gel embeded conditions, CTOS-derived xenograft tumors, and corresponding microdissected patient tumors. One experiment was performed for each sample.
Contributor(s) Inoue M
Citation(s) 27098764
Submission date Dec 29, 2015
Last update date Nov 27, 2018
Contact name Masahiro Inoue
Organization name Kyoto Univ Graduate School of Medicine
Department Clinical Bio-resource Research and Development
Street address Med-Pharm Collaboration Bldg 503, Shimoadachi-cho 46, Sakyou-ku, Kyoto, 606-8304, Japan
City Kyoto
State/province Kyoto
ZIP/Postal code 606-8304
Country Japan
Platforms (1)
GPL13607 Agilent-028004 SurePrint G3 Human GE 8x60K Microarray (Feature Number version)
Samples (32)
GSM1986853 C45 CTOS_in gel
GSM1986854 C45 CTOS_in suspension
GSM1986855 C45 CTOS_original tumor
BioProject PRJNA307193

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE76405_RAW.tar 401.1 Mb (http)(custom) TAR (of TXT)
Processed data included within Sample table

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