This is a collaborative project between

Laboratory of Tumor Immunology and Biology, Center for Cancer Research, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA.

Biomedical Center, StPeterburgh, Russia

Molecular and Microbiology Department, College of Science,George Mason University, Fairfax, VA

This study has been published: Clin Cancer Res. 2007 Apr 15;13(8):2471-8.

Purpose: Identification of tumor antigens is essential in advancing immune-based therapeutic interventions in cancer. Particularly attractive targets are those molecules that are selectively expressed by malignant cells and that are also essential for tumor progression.

Experimental Design and Results: We have used a computer-based differential display analysis tool for mining of expressed sequence tag clusters in the human Unigene database and identified Brachyury as a novel tumor antigen. Brachyury, a member of theT-box transcription
factor family, is a key player inmesodermspecification during embryonic development.Moreover, transcription factors that control mesoderm have been implicated in the epithelial-mesenchymal transition (EMT), which has been postulated to be a key step during tumor progression to metastasis. Reverse transcription-PCR analysis validated the in silico predictions and showed Brachyury expression in tumors of the small intestine, stomach, kidney, bladder, uterus, ovary, and testis, as well as in cell lines derived from lung, colon, and prostate carcinomas, but not in the vast majority of the normal tissues tested. An HLA-A0201epitope of human Brachyury was identified that was able to expand T lymphocytes from blood of cancer patients and normal donors with the ability to lyse Brachyury-expressing tumor cells.

Conclusions: To our knowledge, this is the first demonstration that (a) a T-box transcription factor and (b) a molecule implicated in mesodermal development, i.e., EMT, can be a potential target for human T-cell mediated cancer immunotherapy.

Follow-up study was published in: Vopr Onkol. 2008;54(6):739-43.

Investigation of transcription factor Brachyury (T) expression in human normal and tumor tissues

Krukovskaia LL, Polev DE, Nosova IuK, Baranova AV, Koliubaeva SN, Kozlov AP.

Follow-up study: By using computational differential display approach we identified a number of UniGene clusters which comprised 90% or more of ESTs from tumor cDNA libraries. One of them was cluster Hs.389457 which corresponds to the human gene Brachyury (T). That encodes a T-box gene family member transcription factor which is pivotal in early embryonal development. To experimentally verify our in silico findings of T expression, PCR was conducted using panels of cDNA from various human normal and tumor tissues. According to our results, Brachynry is expressed in tumors of the digestive tract, testis, ovary, breast, kidney, bladder, lung and brain tunic as well as in lymphomas. Weak amplification signals were picked up from normal tissues of small intestine, spleen and testis. Our results support earlier hypothesis on predominant tumor-related expression of Brachyury gene in adults.