Transcriptome Analysis of Tumor-Infiltrating Lymphocytes Identifies NK Cell Gene Signatures Associated With Lymphocyte Infiltration and Survival in Soft Tissue Sarcomas

Abstract

PurposeClinical successes using current T-cell based immunotherapies have been limited in soft tissue sarcomas (STS), while pre-clinical studies have shown evidence of natural killer (NK) cell activity. Since tumor immune infiltration, especially tumor-infiltrating lymphocytes, is associated with improved survival in most solid tumors, we sought to evaluate the gene expression profile of tumor and blood NK and T cells, as well as tumor cells, with the goal of identifying potential novel immune targets in STS.Experimental DesignUsing fluorescence-activated cell sorting, we isolated blood and tumor-infiltrating CD3-CD56+ NK and CD3+ T cells and CD45- viable tumor cells from STS patients undergoing surgery. We then evaluated differential gene expression (DGE) of these purified populations with RNA sequencing analysis. To evaluate survival differences and validate primary DGE results, we also queried The Cancer Genome Atlas (TCGA) database to compare outcomes stratified by bulk gene expression.ResultsSorted intra-tumoral CD3+ T cells showed significant upregulation of established activating (CD137) and inhibitory genes (TIM-3) compared to circulating T cells. In contrast, intra-tumoral NK cells did not exhibit upregulation of canonical cytotoxic genes (IFNG, GZMB), but rather significant DGE in mitogen signaling (DUSP4) and metabolic function (SMPD3, SLC7A5). Tumors with higher NK and T cell infiltration exhibited significantly increased expression of the pro-inflammatory receptor TLR4 in sorted CD45- tumor cells. TCGA analysis revealed that tumors with high TLR4 expression (P = 0.03) and low expression of STMN1 involved in microtubule polymerization (P < 0.001) were associated with significantly improved survival.ConclusionsUnlike T cells, which demonstrate significant DGE consistent with upregulation of both activating and inhibiting receptors in tumor-infiltrating subsets, NK cells appear to have more stable gene expression between blood and tumor subsets, with alterations restricted primarily to metabolic pathways. Increased immune cell infiltration and improved survival were positively correlated with TLR4 expression and inversely correlated with STMN1 expression within tumors, suggesting possible novel therapeutic targets for immunotherapy in STS.

Comprehensive analysis of cuproptosis-related lncRNAs in immune infiltration and prognosis in hepatocellular carcinoma

“…NK cells fall under the cell type of the innate lymphoid branch. Tumorous immunological infiltrate, especially tumor-infiltrating lymphocytes, is linked to better survival in the majority of solid cancers [56][57][58]. It is generally assumed that low levels of NK cell infiltration and NK cell dysfunction are connected with worse OS as well as tumor recurrence following therapy [58,59].…”

“…Tumorous immunological infiltrate, especially tumor-infiltrating lymphocytes, is linked to better survival in the majority of solid cancers [56][57][58]. It is generally assumed that low levels of NK cell infiltration and NK cell dysfunction are connected with worse OS as well as tumor recurrence following therapy [58,59]. Therefore, cuproptosis-related lncRNAs can impact the fate of HCC patients by affecting tumor immune infiltration.…”

Comprehensive Analysis of Cuproptosis-Related lncRNAs in Immune Infiltration and Prognosis in Hepatocellular Carcinoma

“…Tumorous immunological in ltrate, especially tumor-in ltrating lymphocytes, is linked to better survival in the majority solid cancers (38-40). It is generally assumed that low levels of NK cell in ltration and NK cell dysfunction are connected with worse OS as well as tumor recurrence following theapy (40,41). Therefore, cuproptosis-related lncRNAs can impact the fate of HCC patients by affecting tumor immune in ltration.…”