Type: Oral
Session: 651. Multiple Myeloma and Plasma Cell Dyscrasias: Basic and Translational: Microenvironment and Immunity in Myeloma
Hematology Disease Topics & Pathways:
Research, Fundamental Science, Translational Research, Plasma Cell Disorders, Bioinformatics, Diseases, Immunology, Lymphoid Malignancies, Computational biology, Biological Processes, Technology and Procedures
Aim. To define the contribution of tumour features to T cell differentiation during disease evolution from precursor disease to MM.
Methods. We generated the “pre-myeloma atlas”, the largest single-cell RNA sequencing (scRNA-seq) dataset of precursor disease and MM to date. It encompassed >1,000,000 cells from 295 newly-generated and published scRNA-seq samples from MM (n=54 patients), precursor conditions (n=78), and non-cancer controls (n=102). We enhanced the atlas through including single-cell T cell receptor (TCR) sequencing (42% samples), bone marrow and peripheral blood sampling, and modelling clinical data including age and tumour burden. Malignant clones (tumour cells) were identified by clonal immunoglobulin expression and scored with pan-cancer transcriptional pathways. Viral-reactive TCRs were acquired from VDJdb and neoantigen-reactive TCRs predicted with NetMHCpan4.1 and TEINet.
Results. Within the pre-myeloma atlas, we identified CD4+ and CD8+ T cell subsets across the spectrum of differentiation from naïve (SELL+CCR7+) to terminal memory (GZMB+) phenotypes. Notably, we identified exhausted T cells in only a single MM patient (1 of 54), suggesting T cell exhaustion is not a pervasive feature of MM. Comparing controls with precursor and symptomatic patients, we found T cell composition was comparable through disease evolution. However, the abundance of T cell subsets was altered: in MM, earlier differentiated subsets such as naïve CD8+ were depleted relative to controls (P<0.001) whereas terminal memory subsets were enriched (P<0.003). Principal component analysis of T cell composition captured this shift from early to terminal subsets, which we termed T cell skewing. T cell skewing rose with disease severity and was enhanced in SMM and MM independent of patient age (SMM P=0.004; MM P<0.001). Longitudinal sampling of precursor patients who subsequently progressed revealed T cell skewing occurs longitudinally at progression. Together, this suggests exaggerated T cell memory differentiation occurs in MM.
TCR repertoire clonality (indicative of T cell proliferation in response to antigen) was higher in MM relative to controls (P=0.06) and corrected with T cell skewing (R=0.71, P<0.001). Additionally, the frequency of TCR clones possessing similar antigen-recognition domains (indicative of shared antigen-specificity) rose between SMM and MM (P<0.002) and correlated with T cell skewing (R=0.4, P=0.02). Importantly, in silico analyses of T cell specificity suggested (1) TCR clones annotated as viral-reactive were not associated with T cell skewing, and (2) TCR clones predicted to bind autologous tumour neoantigens occupied terminal memory states. Together, these results suggest antigen, possibly tumour in origin, drive T cell differentiation in MM.
Finally, to interrogate mechanisms of tumour-immune cross-talk, we compared T cell and tumour features. We observed a positive correlation between T cell skewing with paraprotein (R=0.43, P=0.04), that serves as a proxy for tumour burden. Additionally, effector T cells (IFNG+TNF+) were associated with tumour marrow infiltration (aspirate % CD138+; R=055, P<0.001). When analysing the transcriptional activity of tumour cells, we found effector T cells also correlated with cell death-associated pathways (R=0.66, P<0.001). We noted sharing of TCRs between these two T cell subsets, suggesting a differentiation process between these subsets possibly regulated by tumour-intrinsic processes and tumour burden.
Conclusion. Our results show that, unlike solid cancers, MM is not associated with T-cell exhaustion and instead defined by a pattern of T-cell differentiation resembling antigen-driven terminal memory differentiation. T cell associations with tumour burden and antigen could suggest anti-tumour immunity drives a novel form of cancer-associated T-cell differentiation in MM.
Disclosures: Walker: Abbvie: Consultancy. Ramasamy: Pfizer: Consultancy, Speakers Bureau; Recordati rare Disease: Consultancy, Speakers Bureau; Menarini Stemline: Consultancy, Speakers Bureau; Johnson and Johnson: Consultancy, Speakers Bureau; Sanofi: Consultancy, Research Funding, Speakers Bureau; Takeda: Consultancy, Speakers Bureau; GSK: Consultancy, Research Funding, Speakers Bureau; Bristol Myers Squibb: Consultancy, Research Funding, Speakers Bureau; Amgen: Consultancy, Research Funding, Speakers Bureau; Adaptive Biotech: Consultancy, Speakers Bureau. Herrero: Asgard Therapeutics: Current Employment. Quezada: Achille Theraputics: Current Employment.