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Alona Lanksy



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    P2.04 - Immuno-oncology (ID 167)

    • Event: WCLC 2019
    • Type: Poster Viewing in the Exhibit Hall
    • Track: Immuno-oncology
    • Presentations: 1
    • Moderators:
    • Coordinates: 9/09/2019, 10:15 - 18:15, Exhibit Hall
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      P2.04-04 - CITEseq Characterization in Early Stage NSCLC Patients Identifies Distinct Patterns of Immune Infiltrate (ID 2305)

      10:15 - 18:15  |  Author(s): Alona Lanksy

      • Abstract

      Background

      The success of immunotherapy in late-stage lung cancer patients, together with the need for novel therapies for early-stage disease, mandates an increased understanding of the immune infiltrate in early-stage lesions. Recent advances in sequencing-based single-cell technologies have enabled an unprecedented degree of resolution in the phenotypic characterization of patient tissues.

      Method

      Tumor and non-involved lung resection specimens were acquired from 23 early-stage NSCLC patients. Immune cells were isolated and analyzed by single-cell RNAseq (scRNAseq) using the 10X Chromium platform. Resulting expression signatures were clustered using an in-house pipeline. To validate populations and elucidate surface marker staining patterns for transcriptionally-defined cell clusters, we used cellular indexing of transcriptomes and epitopes by sequencing (CITEseq)—using oligonucleotide-conjugated antibodies to simultaneously measure expression of over 50 surface proteins along with transcriptomes of single cells—to analyze tumors from 8 additional patients. To identify T cell phenotypes that were differentially present and clonally expanded within tumor compared to non-involved lung, we paired scRNAseq with T cell receptor repertoire profiling in 3 patients. Finally, to validate the transcriptional phenotypes we detected and to extend our dataset, we incorporated 8 patients from a public dataset, totaling 39 patients included in the study. Immune signatures were correlated with presence of actionable mutations, smoking history, stage, and histology.

      Result

      Using these single cell analyses, all major immune cell lineages were identified within tumors, including multiple distinct myeloid and lymphoid subsets, which notably were phenotypically distinct from those isolated from uninvolved lung tissue. Existing databases of ligand-receptor pairs were leveraged to construct an interactome, implicating specific axes of cell-cell communication in driving changes common to tumors. As we hypothesized, correlative analyses across tumor samples revealed a cellular module marked by exhausted T cells, plasma cells, mature dendritic cells, and monocyte-derived macrophages that was enriched in patients with significant smoking histories and EGFRWT disease.

      Conclusion

      These findings indicate that strong immune differences exist between treatment-naïve lesions, and that these differences stratify with smoking history and smoking-related driver mutations. Given existing literature indicating that positive smoking history confers improved response to immune checkpoint blockade, our data suggests that this disparity may be mediated by set differences in treatment-naïve immune microenvironments. We will now apply this analysis pipeline to tumors treated in the neoadjuvant setting in an ongoing trial (submitted to WCLC in abstract form).