Abstract 5773: Sequencing and cloning IDH1 R132H-targeted monoclonal T cell receptors from CD4+ T cells facilitated by opto-electric-positioning technology

• Published in: Cancer research (Chicago, Ill.) Ill.), 2018-07, Vol.78 (13_Supplement), p.5773-5773
• Main Authors: Smith, Duane, Watchmaker, Payal, Stadler, Guido, Marks, Natalie, Bronevetsky, Yelena, Chapman, Keviin, Okada, Hideho
• Format: Article
• Language: English
• ISSN: 0008-5472; 1538-7445
• DOI: 10.1158/1538-7445.AM2018-5773

Isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) are enzymes that catalyze the conversion of isocitrate to α-ketoglutarate, while reducing NADH to NADPH. More than 80% of low-grade gliomas and secondary glioblastomas harbor point mutations in IDH. The R132H mutation is the most common mutation in IDH1, and vaccination with the mutant peptide elicited an MHC class II-specific antitumor response, suggesting mutant IDH1 as a potential target for immunotherapy (1). Discovering monoclonal T cell receptors (TCRs) that specifically recognize the R132H mutation is a key but challenging step towards viable immunotherapy for glioma/glioblastomas. Using opto-electric-positioning (OEP) technology in a chip-based, NanoPen culture environment, we have successfully identified and isolated a monoclonal T cell that specifically recognizes the R132H mutant IDH1 peptide. We cultured and identified monoclonal CD4+ T cells on chip, stimulated by monocyte-derived dendritic cells, pulsed with R132H IDH1 peptide. Activated, CD4+ T cells were selectively exported from the chip and their T-cell receptors (TCRs) were subsequently deep sequenced. The most likely pairs of alpha and beta TCR chains were cloned into mammalian expression vectors for functional validation in vitro, producing a TCR that specifically binds mutant IDH1 tetramer. Taken together, we show that the combined platform of the OEP and nanofluidic device provides a powerful solution to help identify cell type-specific, monoclonal T cells that can be exported and sequenced to reveal distinct TCRs, which in turn can be cloned, expressed and validated for future immunotherapies. Reference: 1. Schumacher et al., Nature 2014;512:324-27. Citation Format: Duane Smith, Payal Watchmaker, Guido Stadler, Natalie Marks, Yelena Bronevetsky, Keviin Chapman, Hideho Okada. Sequencing and cloning IDH1 R132H-targeted monoclonal T cell receptors from CD4+ T cells facilitated by opto-electric-positioning technology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5773.