A strategy for uncovering germline variants altering anti-tumor CD8 T cell response

Vijay Kumar Ulaganathan; Martina H. Vasileva

doi:10.1016/j.jgg.2023.01.001

Volume 50 Issue 5

May 2023

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Article Navigation > Journal of Genetics and Genomics > 2023 > 50(5): 353-361

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A strategy for uncovering germline variants altering anti-tumor CD8 T cell response

doi: 10.1016/j.jgg.2023.01.001

Vijay Kumar Ulaganathan^a,b,c,
Martina H. Vasileva^a

a. Klinik für Dermatologie & Allergology, Universitätsmedizin Göttingen, Göttingen 37075, Germany;
b. Institut für Multiple Sklerose Forschung, Neuroimmunologie, Universitätsmedizin Göttingen, Göttingen 37075, Germany;
c. University of Lorraine, NGERE Unit, Faculté de Médecine, 9 Avenue de La Forêt de Haye, Vandoeuvre-lès-Nancy 54505, France

Funds:

rner for helpful discussions and advice. We thank Simon Mole, Simone Hamann, Birgitte Salzmann, Birgitte Curdt, Martina Weig and Karolin Zachmann for the technical support.

hder for providing the OT-I mice, Marc-André

The authors thank Ambry Genetics for providing aggregated and annonymized germline genotyping data through the “AmbryShare” Program, Alexander Flü

gel, Michael P. Schö

Lé

cuyer for flow cytometry and comments on the manuscript, Francesca Odoardi, Dimitri Lodygin and Henrike Kö

n, Jorge F Alberto, Jean-Louis Gueant, David Meyre, Jean-Marc Alberto, for supporting this work, Fred Lü

Received Date: 2022-10-28
Accepted Date: 2023-01-03
Rev Recd Date: 2022-12-22
Publish Date: 2023-01-20

Abstract

Abstract

Among many factors known to alter the outcomes of T cell receptor (TCR)-induced proximal signaling, the role of human germline variants in dictating the individuality of the anti-tumor CD8 T cell response has remained challenging to address. Here, we describe a convenient strategy for molecular and functional characterization of phosphotyrosine-altering non-synonymous single nucleotide variations (pTyr-SNVs) that directly impact TCR-induced proximal phosphotyrosine motif-based signaling pathways. We devise an experimental co-cultivation set-up comprising a C57BL/6 mouse-derived metastatic melanoma cell line engineered to constitutively present ovalbumin (OVA) antigens and retrovirally engineered syngeneic major histocompatibility complex (MHC) Class I restricted OVA TCR-transgenic CD8 T cells (OT-I). Using the synthetic version of pTyr-SNV rs1178800678-G/T-encoding integrin alpha 4 (ITGA4) p.S1027I variant as a prototype, we show that under identical TCR stimulation conditions, genetically determined membrane-proximal immunoreceptor tyrosin activation motif (ITAM) results in increased tyrosine phosphorylation of 70 kDa zeta-chain-associated protein (ZAP70) and the levels of cytotoxic effector molecule granzyme B (GZMB), which in turn result in enhanced cytotoxic activity against metastatic melanoma cell line. This strategy paves the way for rapid molecular and functional characterization of anti-tumor immune response-linked germline pTyr-SNVs so as to improve our understanding of the genetic basis of individual-to-individual differences in anti-tumor CD8 T cell response.

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