Poster Exhibition Area
Zelluna Immunotherapy AS
Generation and characterization of TCR-NK cells for solid cancer therapy
In recent years, emerging natural killer (NK) cell therapies have shown promise in initial clinical trials, due to their efficacy in killing tumour, as well as their favourable safety profile showing no evidence of cytokine release syndrome or neurotoxicity. The use of NK cells has the potential to overcome limitations related to time and cost of manufacturing and to the quality of the final product compared to autologous cell therapies as they can be used in an allogeneic setting, as off-the-shelf treatments. TCR based therapies have shown compelling data demonstrating effective targeting of solid tumours and clinical responses across numerous solid cancers. Therefore, with our proprietary TCR-NK platform, we exploit the solid tumour targeting capabilities of TCRs with the cytotoxic potential of highly potent killer cells, redirecting and arming NK cells with a fully functional T cell receptor (TCR)-CD3 complex.
We generated TCR-NK cells from different cell sources, using peripheral blood NK, cord blood NK and CD34-positive hematopoietic stem cells from umbilical cord blood as starting cell material. We transduced NK cells, or CD34-positive stem cells cultured ex vivo and differentiated into CD56+ NK cells, with a tool cancer testis antigen (CTA) specific TCR as well as CD3 and optionally a CD8 co-receptor. Importantly, the introduction of the TCR did not alter the activating and inhibitory receptor profile of the generated NK cells. We show that the TCR-NK cells were able to induce antigen specific TCR-driven cytotoxicity and degranulation when presented with cancers expressing natural levels of antigen while maintaining their innate recognition and killing function. Taken together, this data demonstrates the feasibility of TCR-NK generation from different starting cell populations, as well as a beneficial effect of the TCR, redirecting NK cells to natural antigen expressing targets and showing TCR-mediated killing in addition to the innate killing function. This data underlines the therapeutic potential of a novel class of treatment for solid malignancies, namely “off the shelf” TCR-NK cell therapies.
Sylvie Pollmann, Ines Cardoso, Margherita Boieri, Eva-Kristine Klemsdal, Emmanuelle Benard, Anja Oldenburg, Amanda Ruud, Julia Ino, Namir Hassan, Luise U. Weigand.
Zelluna Immunotherapy AS, Ullernchausseen 64, 0379 Oslo
Children's Hospital of Philadelphia & University of Pennsylvania
High throughput pMHC-I tetramer library production using chaperone-mediated peptide exchange
Peptide exchange technologies are essential for the generation of pMHC-multimer libraries used to probe diverse, polyclonal TCR repertoires in various settings. Here, using the molecular chaperone TAPBPR, we develop a robust method for the capture of stable, empty MHC-I molecules comprising common human HLA-A and B alleles, which can be readily tetramerized and loaded with peptides of choice in a high-throughput manner. Alternatively, catalytic amounts of TAPBPR can be used to exchange placeholder peptides with high affinity peptides of interest. Using the same system, we describe high throughput assays to validate binding of multiple candidate peptides on empty MHC-I/TAPBPR complexes. Combined with tetramer-barcoding via a multi-modal cellular indexing technology, ECCITE-seq, our approach allows a combined analysis of TCR repertoires and other T cell transcription profiles together with their cognate antigen specificities in a single experiment. The new approach allows TCR/pMHC interactions to be interrogated easily at large scale.
Yi Sun, Nikolaos G. Sgourakis (corresponding author)
Center for Computational and Genomic Medicine, Department of Pathology and Laboratory Medicine, The Children’s Hospital of Philadelphia & Department of Biochemistry and Biophysics, Perelman School of Medicine, University of Pennsylvania
Jugmohit S. Toor*, Sarah A. Overall* (*co-first authors), Sarah M. O’Rourke, Danai Moschidi, Giora I. Morozov, Nicolas Gonzalez
Department of Chemistry and Biochemistry, University of California Santa Cruz, 1156 High St., Santa Cruz, CA, 95064, USA
Mark Yarmarkovich, John M. Marris
Division of Oncology, Center for Childhood Cancer Research, Children’s Hospital of Philadelphia and Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania, 3400 Civic Center Blvd, Philadelphia, PA, 19104, USA
Alberto S. Japp, Son Nguyen, Michael R. Betts
Department of Microbiology, Perelman School of Medicine, University of Pennsylvania, 3400 Civic Center Blvd, Philadelphia, PA, 19104, USA
Peter Smibert, Stephanie Hao
Technology Innovation Lab, New York Genome Center, 101 6th Ave, New York, NY, 10013, USA
TCR-Ts expressing chimeric PD1-41BB and a highly potent PRAME-TCR show enhanced anti-tumor efficacy
Using a high-throughput TCR-generation process, a highly specific TCR recognizing the C/T antigen PRAME with high sensitivity was identified. To improve TCR-T function in solid tumor environments the TCR lead candidate was co-expressed with a chimeric PD1-41BB co-stimulatory receptor. PD1-41BB-expressing TCR-Ts retained a favorable preclinical safety profile and showed enhanced anti-tumor reactivity in vitro. This potent anti-tumor efficacy could be confirmed in a xenograft mouse model, where tumors were eradicated in vivo by a single dose of TCR-Ts co-expressing PD1-41BB and the PRAME-TCR.
Nadja Sailer1, Ina Fetzer1, Melanie Salvermoser1, Doris Brechtefeld1, Maja Bürdek1, Kathrin Mutze1, Monika Braun1, Dolores J. Schendel2, Susanne Wilde1, Daniel Sommermeyer1
1Medigene Immunotherapies GmbH, a subsidiary of Medigene AG, Planegg/Martinsried, Germany; 2Medigene AG, Planegg/Martinsried, Germany