Globo H expression in metastatic colorectal cancer (CRC).
Authors: Jayachandran Priya , Baca Yasmine , Xiu Joanne , Zhang Jian , Battaglin Francesca , Arai Hiroyuki , Goldberg Richard M. , Weinberg Benjamin Adam , Lou Emil , Hall Michael J. , Khushman Moh'd M. , Sohal Davendra , Soni Shivani , Wang Jingyuan , Zhang Wu , Millstein Joshua , Korn Wolfgang Michael , Lenz Heinz-Josef 
Published: June 2021

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Background: Globo H is a carbohydrate antigen that is highly expressed on the cell surface of epithelial cancers but not in normal tissue, and has been reported to correlate with poor prognosis. An attractive therapeutic target, Globo H-targeted agents are being tested in early clinical trials (e.g., OBI-833, a Globo H antigen conjugated to a mutated diphtheria toxin with potential antineoplastic activities, and OBI-999, an antibody-drug conjugate (ADC) consisting of a Globo H monoclonal antibody with a synthetic antineoplastic agent). We aim to describe the molecular features associated with Globo H expression in CRC. Methods: A total of 7,604 CRC tumors were tested by Caris Life Sciences (Phoenix, AZ) by NextGen DNA and RNA sequencing. The expression of β3GalT5, FUT-1 and FUT-2 were evaluated as surrogates for Globo H expression as they are the key enzymes in its biosynthesis. An average z-score of the 3 genes (GloboH) and of β3GalT5 (B3) alone were calculated; tumors with top quartile z-scores were considered expression-high (Q4) and bottom quartile, expression-low (Q1). QuantiSEQ was used to assess immune cell infiltration in the tumor microenvironment (TME). Statistical significance was determined using chi-square/Fisher-Exact and adjusted for multiple comparisons (q<0.05). Consensus molecular subtype (CMS) was developed using RNA seq data. Results: When the 3 genes were considered, GloboH-H tumors showed higher prevalence of CMS1 and CMS4 (23.8% vs. 12%; 38.7% vs. 29.4%) and lower prevalence of CMS2 (40% vs. 18.7%) compared to GloboH-L. Similar patterns of CMS distribution were seen for B3 alone. B3-H tumors were significantly more frequently TMB-H (>=10) (11.4% vs. 8.3%), PD-L1 positive (5.7% vs. 3.4%) and MSI-H/dMMR (8.3% vs. 5.5%). Strong positive associations were seen with mutations in BRAF, KRAS, RSPO3 fusion, and cMYC amplification with B3 alone and GloboH (all q<0.05). Anti-tumor CD4+ T cells and NK cells were increased in the TME with increased expression of GloboH and B3 (q<0.05). However, immune suppressive neutrophils and Tregs were also increased. Dendritic cells were negatively associated with B3 expression while endothelial cells and fibroblasts showed a positive association with GloboH and B3. Conclusions: The association with TMB-H, MSI-H, and PD-L1 status suggests that in some tumors Globo H may be a promising target for combination therapy with immune checkpoint inhibition. The association with different cell populations suggests manipulating the cellular balance in the TME as an approach to improve the efficacy of treatment. NK cell checkpoint inhibitors are in clinical trials and might be utilized in high Globo H cancers; treatments inducing DCs in tumors have been shown to enhance responses to BRAF and PD-L1 blockade and might be applicable in the context of Globo H immunotherapy to overcome Treg immune suppression. Anti-Globo H vaccines and ADCs might be particularly effective in BRAF and KRAS-mutant CRC patients.