Anti-sialyl-di-lewis^a CAR T cells for effective anti-tumour therapy

Iniobong Charles, Katherine Cook, Bubacarr Kaira, Gaëlle Cane, Anne Skinner, Alissa Wright, Mireille Vankemmelbeke, Rachael Metheringham, Victoria Brentville and Lindy Durrant

Unlocking the unique potential of AvidiMAb® in fighting cancer

Bubacarr G Kaira, Foram Dave, Elena Dubinina, Omar Mohammed, Bryony Heath, Poonam Vaghela, Ruhul Choudhury, Tina Parsons, Smantha Paston and Mireille Vankemmelbeke and Lindy G Durrant

GlyMabs, glycome-targeting monoclonal antibodies for cancer therapy

Ruhul H Choudhury, Omar Mohammed, Foram Dave, Bubacarr Kaira, Poonam Vaghela, Bryony Heath, Elena Dubinina, Tina Parsons, Mireille Vankemmelbeke and Lindy  G Durrant

An Immunohistochemical Evaluation of Tumor-Associated Glycans and Mucins as Targets for Molecular Imaging of Pancreatic Ductal Adenocarcinoma

Ruben D. Houvast, Kira Thijse, Jesse V. Groen, JiaXin Chua, Mireille Vankemmelbeke, Lindy G. Durrant, J. Sven D. Mieog, Bert A. Bonsing, Alexander L. Vahrmeijer, Peter J.K. Kuppen, A. Stijn L.P. Crobach and Cornelis F. M. Sier

SIMPLE SUMMARY: Distinguishing pancreatic cancer from healthy tissue before and during surgery can be enhanced by using molecular tracers directed at molecules on tumor cells allowing highcontrast visualization of tumor tissue, eventually improving diagnosis and surgical removal. Albeit sugar molecules and proteins carrying a large amount of sugars-mucins- have gained significant interest as tumor-specific targets, their relative presence on structures surrounding tumor tissues and lymph node metastases is unknown. The current study shows that the presence of several, but not all, investigated sugar molecules and mucins on pancreatic cancer cells is higher compared to surrounding tissues. Moreover, given their abundance on tumor cells in lymph nodes and their absence on normal lymph nodes, all investigated targets are high-potential targets for visualization of lymph node metastases. This study paves the way for the development of molecular tracers against the targets evaluated herein to allow improvement of pancreatic cancer treatment.

ABSTRACT: Targeted molecular imaging may overcome current challenges in the preoperative and intraoperative delineation of pancreatic ductal adenocarcinoma (PDAC). Tumor-associated glycans Lea/c/x, sdi-Lea, sLea, sLex, sTn as well as mucin-1 (MUC1) and mucin-5AC (MU5AC) have gained significant interest as targets for PDAC imaging. To evaluate their PDAC molecular imaging potential, biomarker expression was determined using immunohistochemistry on PDAC, (surrounding) chronic pancreatitis (CP), healthy pancreatic, duodenum, positive (LN+) and negative lymph node (LN¯) tissues, and quantified using a semi-automated digital image analysis workflow. Positive expression on PDAC tissues was found on 83% for Lea/c/x, 94% for sdi-Lea, 98% for sLea, 90% for sLex, 88% for sTn, 96% for MUC1 and 67% for MUC5AC, where all were not affected by the application of neoadjuvant therapy. Compared to PDAC, all biomarkers were significantly lower expressed on CP, healthy pancreatic and duodenal tissues, except for sTn and MUC1, which showed a strong expression on duodenum (sTn tumor:duodenum ratio: 0.6, p < 0.0001) and healthy pancreatic tissues (MUC1 tumor:pancreas ratio: 1.0, p > 0.9999), respectively. All biomarkers are suitable targets for correct identification of LN+, as well as the distinction of LN+ from LN¯ tissues. To conclude, this study paves the way for the development and evaluation of Lea/c/x-, sdi-Lea-, sLea-, sLex- and MUC5AC-specific tracers for molecular imaging of PDAC imaging and their subsequent introduction into the clinic.

Targeting Glycans and Heavily Glycosylated Proteins for Tumor Imaging

Ruben D. Houvast, Mireille Vankemmelbeke, Lindy G. Durrant, Manfred Wuhrer, Victor M. Baart, Peter J.K. Kuppen, Lioe-Fee de Geus-Oei, Alexander L. Vahrmeijer and Cornelis F.M. Sier

SIMPLE SUMMARY:  Distinguishing malignancy from healthy tissue is essential for oncologic surgery. Targeted imaging during an operation aids the surgeon to operate better. The present tracers for detecting cancer are directed against proteins that are overexpressed on the membrane of tumor cells. This review evaluates the use of tumor-associated sugar molecules as an alternative for proteins to image cancer tissue. These sugar molecules are present as glycans on glycosylated membrane proteins and glycolipids. Due to their location and large numbers per cell, these sugar molecules might be better targets for tumor imaging than proteins.

Monoclonal Antibody Targeting Sialyl-di-Lewis^a – Containing Internalizing and Noninternalizing Glycoproteins with Cancer Immunotherapy Development Potential

Silvana T. Tivadar, Richard S. McIntosh, Jia Xin Chua, Robert Moss, Tina Parsons, Abed M. Zaitoun, Srinivasan Madhusudan, Lindy G. Durrant and Mireille Vankemmelbeke

ABSTRACT: Tumor glycans constitute attractive targets for therapeutic antibodies. The sialylated glycocalyx plays a prominent role in cancer progression and immune evasion. Here, we describe the characterization of the mAb, FG129, which targets tumor-associated sialylated glycan, and demonstrate its potential for multimodal cancer therapy. FG129, obtained through BALB/c mouse immunizations with liposomes containing membrane glycan extracts from the colorectal cancer cell line LS180, is an mIgG1k that targets sialyl-di-Lewis^a–containing glycoproteins. FG129, as well as its chimeric human IgG1 variant, CH129, binds with nanomolar functional affinity to a range of colorectal, pancreatic, and gastric cancer cell lines. FG129 targets 74% (135/182) of pancreatic, 50% (46/92) of gastric, 36% (100/281) of colorectal, 27% (89/327) of ovarian, and 21% (42/201) of non–small cell lung cancers, by IHC. In our pancreatic cancer cohort, high FG129 glyco-epitope expression was significantly associated with poor prognosis (P ¼ 0.004). Crucially, the glyco-epitope displays limited normal tissue distribution, with FG129 binding weakly to a small percentage of cells within gallbladder, ileum, liver, esophagus, pancreas, and thyroid tissues. Owing to glyco-epitope internalization, we validated payload delivery by CH129 through monomethyl auristatin E (MMAE) or maytansinoid (DM1 and DM4) conjugation. All three CH129 drug conjugates killed high-binding colorectal and pancreatic cancer cell lines with (sub)nanomolar potency, coinciding with significant in vivo xenograft tumor control by CH129-vcMMAE. CH129, with its restricted normal tissue distribution, avid tumor binding, and efficient payload delivery, is a promising candidate for the treatment of sialyl-di-Lewis^a– expressing solid tumors, as an antibody–drug conjugate or as an alternative cancer immunotherapy modality.

An ultraspecific anti-SSEA-4 monoclonal antibody recognizes stem memory T cells

J Chua, E Cid, M Vankemmelbeke, R McIntosh, R Metheringham, I Daniels, V Brentville and L Durrant

Induction of post-translational modifications in tumour and their recognition by T cells

P Bilimoria, R Metheringham, M Gijon, V Brentville, K Cook, P Symonds, I Daniels and L Durrant