Dados do Resumo

Título

CD56 bright NK cell is important driver involved in the maturation of immune hubs and better prognosis of patients with clear cell renal cell carcinoma

Introdução

The understanding of the immune infiltrate and its organization within the tumor microenvironment (TME) has been transforming cancer management. Tumor-associated tertiary lymphoid structures (TLS), have been linked to improved prognosis in tumors such as clear cell renal cell carcinoma (ccRCC). However, the interaction between TLS and natural killer (NK) cells, an important cell with anti-tumor activity, remains poorly understood. Comprehensive characterization of NK cells within the TME and their interactions with TLS could provide crucial insights for developing new therapeutic strategies for ccRCC.

Objetivo

To characterize the NK cells phenotype in different compartments of patients with ccRCC, evaluate the relationship between the NK cell functionality and the maturation stages of the TLS in patients with ccRCC, as well as to assess prognostic value of NK cell infiltration in ccRCC patients.

Métodos

The study was approved by the Institutional Research Ethics Committee (3223/22). First, we evaluated the NK cell phenotype in tumor, peritumoral and blood samples from treatment-naive ccRCC patients (n = 11) using flow cytometry. We also assessed the spatial organization of immune cells in tumor tissues by immunohistochemistry (IHC) on FFPE tissue slides. To characterize the connection between TLS and NK cell phenotype and gene expression profiles, we performed single-cell RNA sequencing (scRNA-seq) on tumor and peritumoral samples (n = 12). Subsequently, we conducted in vitro experiments to analyze the functionality of NK cells in TLS+ versus TLS- tissues and investigated the mechanisms underlying phenotypic changes in NK cells within TLS+ tumor tissues. Finally, we analyzed bulk RNA-seq and clinical data from ccRCC patients in TCGA to evaluate the clinical relevance of NK cell abundance within the immune infiltrate of these patients.

Resultados

Tumor-derived NK cells presented a distinct phenotype (CD56bright/CD16-), characterized by high expression of resident and activation markers, when compared to those found in blood and peritumor samples (CD56dim/CD16+). Tumor tissue analysis revealed a high density of NK cells within mature TLS (mTLS+), and most of these cells are CD56bright. The scRNA-seq analysis confirmed that the CD16- NK cells are enriched in mTLS+ tumor tissue. Interestingly, CD16- NK cells present within mTLS+tissues, expressed high levels of important chemokines and cytokines (in silico and in vitro), involved in the recruitment and activation of immune cells. Additionally, we found that ccRCC patients showing high expression of CD56 (NK cell marker) and XCR1 (dendritic cell marker) on tumor tissues exhibited improved overall survival when compared to patients with low gene expression of such cell markers.

Conclusões

Our data reveal that ccRCC-derived NK cells exhibit a unique phenotype (CD56bright/CD16-) and display a distinct spatial organization, being closely associated with mTLS. Moreover, we found that CD16- NK cells possess a pronounced capacity to produce high levels of chemokines and cytokines, likely involved in attracting and activating immune cells, such as lymphocytes and dendritic cells. This interaction supports TLS maturation and is linked to improved patient prognosis.

Financiador do resumo

FAPESP, CNPq, CAPES

Palavras Chave

NK cells; Clear cell renal cell carcinoma; tertiary lymphoid structures