Intratumoral NKp46+ natural killer cells are spatially distanced from T and MHC-I+ cells with prognostic implications in soft tissue sarcoma

Cruz, Sylvia M; Sholevar, Cyrus; Judge, Sean J; Darrow, Morgan A.; Iranpur, Khurshid R.; Farley, Lauren E; Lammers, Marshall; Razmara, Aryana; Dunai, Cordelia; Gingrich, Alicia A.; Persky, James; Mori, Hidetoshi; Thorpe, Steven W.; Monjazeb, Arta M.; Murphy, William J.; Canter, Robert J.

doi:10.3389/fimmu.2023.1230534

Cited by 8 publications

(5 citation statements)

References 51 publications

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“…T cells or MHC-I + tumor cells, indicating that the role of NK cells suggested by our results may be context specific (45). This apparent synergy between NK cells and cytotoxic T cells in the NSCLC TME highlights a vital clinical value in developing CPI treatments which stimulate both lymphocyte populations.…”

Section: Cd8 T Cells Had Fewer Mhc-imentioning

confidence: 56%

“…[40][41][42] Our data contrast recent observations in soft tissue sarcoma that NK cell infiltrates indicate a poor prognosis and do not colocalize with CD8 T cells or MHC-I + tumor cells, indicating that the role of NK cells may be context-specific. 43 Our findings that NK cells infiltrated into tumor nests and displayed evidence of IFNγ activity in NSCLC begs the question, why might tumoral NK cells become ineffective drivers of anti-tumor immunity? Emerging evidence suggests that NK cell activity may be hindered in the lung cancer TME.…”

Section: Discussionmentioning

confidence: 87%

“…40–42 Our data contrast recent observations in soft tissue sarcoma that NK cell infiltrates indicate a poor prognosis and do not colocalize with CD8 T cells or MHC-I + tumor cells, indicating that the role of NK cells may be context-specific. 43…”

Section: Discussionmentioning

confidence: 99%

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Spatial colocalization and combined survival benefit of natural killer and CD8 T cells despite profound MHC class I loss in non-small cell lung cancer

Wessel,

Ageeb,

Obeid

et al. 2024

Preprint

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Background.MHC class I (MHC-I) loss is frequent in NSCLC and renders tumor cells resistant to T cell lysis. NK cells kill MHC-I deficient tumor cells, and although previous studies indicate their presence in NSCLC tumor margins, they were also functionally impaired. The goal of this study was to evaluate whether NK cell and CD8 T cell infiltration and activation vary with MHC-I expression.Methods. We used single-stain immunohistochemistry (IHC) and Kaplan-Meier analysis to test the effect of NK cell and CD8 T cell infiltration on overall and disease-free survival. To delineate immune covariates of MHC-I-disparate lung cancers, we used multiplexed immunofluorescence (mIF) imaging followed by multivariate statistical modeling. To identify differences in infiltration and intercellular communication between IFNγ-activated and non-activated lymphocytes, we developed a computational pipeline to enumerate single cell neighborhoods from mIF images followed by multivariate discriminant analysis.Results. Spatial quantitation of tumor cell MHC-I expression revealed intra- and inter-tumoral heterogeneity, which was associated with the local lymphocyte landscape. IHC analysis revealed that high numbers of CD56+cells in patient tumors were associated with disease-free and overall survival (p<0.05). The overall survival association strengthened considerably with high counts of both CD56+and CD8+cells (p<1x10−4). mIF imaging and multivariate discriminant analysis revealed an enrichment of both CD3+CD8+T cells and CD3−CD56+NK cells in MHC-I-bearing tumors (p<0.05). To infer associations of functional cell states and local cell-cell communication, we analyzed spatial single cell neighborhood profiles to delineate the cellular environments of IFNγ+/-NK cells and T cells. We discovered that both IFNγ+NK and CD8 T cells were more frequently associated with other IFNγ+lymphocytes in comparison to IFNγ−NK cells and CD8 T cells (p<1x10−30). Moreover, IFNγ+lymphocytes were most often found clustered near MHC-I+tumor cells.Conclusions. We demonstrate that tumor-infiltrating NK cells and CD8 T cells jointly affect vital control of NSCLC tumor progression. Robust association of NK and CD8 T cells was most evident in MHC-I-bearing tumors, especially in the presence of IFNγ. Frequent co-localization of IFNγ+NK cells with other IFNγ+lymphocytes in near-neighbor analysis indicates lymphocyte activation is coordinately regulated in NSCLC.What is already known on this topic –MHC-I loss occurs frequently in human NSCLC and corresponds with a waning immune response in the tumor microenvironment (TME). NK cells recognize “missing-self” targets and thus may be leveraged to target NSCLC tumors with substantial class I loss. While NK cell presence at tumor margins has been documented in the NSCLC TME, they have been shown to lose function in this environment.What this study adds –We developed spatial statistical analysis pipelines that leverage the local heterogeneity of the tumor microenvironment at single cell resolution to rigorously test whether NK cells and T cells together contribute antitumoral immunity in human lung cancer. We discovered that a high density of tumor-infiltrating NK cells corresponded with disease-free survival, and this association was considerably increased in patients with high coincident T cells, especially those in central tumor. Intriguingly, both cell types were further found clustered together in MHC-I-bearing tumors, especially when both were also expressing IFNγ, thus suggesting coordinated lymphocyte activities may give rise to enhanced immune control of human lung cancer.How this study might affect research, practice, or policy –This study provides a rationale for developing novel immunotherapies that simultaneously increase NK and T cell anti-tumoral immunity. Robust associations linking NK cells with survival and IFNγ activity in NSCLC, including in tumors with profound MHC-I loss, provides a basis for further research to evaluate the effect of NK cells in MHC-I-deficient, CD8 T cell refractory NSCLC tumors.In briefTumor-infiltrated natural killer (NK) and CD8 T cells display increased evidence of co-localization and coordinated effector activity in human MHC-I-bearing NSCLC tumors.

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Section: Cd8 T Cells Had Fewer Mhc-imentioning

confidence: 56%

Section: Discussionmentioning

confidence: 87%

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Spatial colocalization and combined survival benefit of natural killer and CD8 T cells despite profound MHC class I loss in non-small cell lung cancer

Wessel,

Ageeb,

Obeid

et al. 2024

Preprint

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“…Informed consent was obtained from all patients on an Institutional Review Board-approved protocol. Tumor samples were processed into single-cell suspensions for CSC phenotyping and ex vivo exposure to TKIs as described previously [ 19 , 30 , 39 , 48 , 49 ].…”

Section: Methodsmentioning

confidence: 99%

Low-Dose Sorafenib Promotes Cancer Stem Cell Expansion and Accelerated Tumor Progression in Soft Tissue Sarcomas

Cruz,

Iranpur,

Judge

et al. 2024

IJMS

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The cancer stem cell (CSC) hypothesis postulates that heterogeneous human cancers harbor a population of stem-like cells which are resistant to cytotoxic therapies, thus providing a reservoir of relapse following conventional therapies like chemotherapy and radiation (RT). CSCs have been observed in multiple human cancers, and their presence has been correlated with worse clinical outcomes. Here, we sought to evaluate the impact of drug dosing of the multi-tyrosine kinase inhibitor, sorafenib, on CSC and non-CSCs in soft tissue sarcoma (STS) models, hypothesizing differential effects of sorafenib based on dose and target cell population. In vitro, human cancer cell lines and primary STS from surgical specimens were exposed to escalating doses of sorafenib to determine cell viability and expression of CSC marker aldehyde dehydrogenase (ALDH). In vivo, ALDHbright CSCs were isolated, exposed to sorafenib, and xenograft growth and survival analyses were performed. We observed that sarcoma CSCs appear to paradoxically respond to the tyrosine kinase inhibitor sorafenib at low doses with increased proliferation and stem-like function of CSCs, whereas anti-viability effects dominated at higher doses. Importantly, STS patients receiving neoadjuvant sorafenib and RT on a clinical trial (NCT00864032) showed increased CSCs post therapy, and higher ALDH scores post therapy were associated with worse metastasis-free survival. These data suggest that low-dose sorafenib may promote the CSC phenotype in STS with clinically significant effects, including increased tumor growth and higher rates of metastasis formation in sarcoma patients.

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“…Individual patient mean values of the triplicate scores were then classified as high or low with reference to the median score for the entire cohort for each marker, as described previously. 24…”

Section: Immunohistochemical (Ihc) Analysismentioning

confidence: 99%

Characterization of natural killer and cytotoxic T‐cell immune infiltrates in pancreatic ductal adenocarcinoma

Persky,

Cruz,

Darrow

et al. 2024

Journal of Surgical Oncology

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Background and ObjectivesPancreatic ductal adenocarcinoma (PDAC) is an aggressive cancer with poor response to systemic therapies, including immunotherapy. Given the immunotherapeutic potential of natural killer (NK) cells, we evaluated intratumoral NK cell infiltrates along with cytotoxic T cells in PDAC to determine their association with patient outcomes.MethodsWe analyzed tumors from 93 PDAC patients treated from 2012 to 2020. Predictor variables included tumor‐infiltrating lymphocytes (TILs), T‐cell markers (CD3, CD8, CD45RO), NK marker (NKp46), and NK inhibitory marker (major histocompatibility complex class I [MHC‐I]) by immunohistochemistry. Primary outcome variables were recurrence‐free survival (RFS) and overall survival (OS).ResultsMean TILs, CD3, and NKp46 scores were 1.3 ± 0.63, 20.6 ± 17.5, and 3.1 ± 3.9, respectively. Higher expression of CD3 and CD8 was associated with higher OS, whereas NK cell infiltration was not associated with either RFS or OS. There was a tight positive correlation between MHC‐I expression and all T‐cell markers, but not with NKp46.ConclusionsOverall NK cell infiltrates were low in PDAC and did not predict clinical outcomes, whereas T‐cell infiltrates did. Further characterization of the immune infiltrate in PDAC, including inhibitory signals and suppressive cell types, may yield better biomarkers of prognosis and immune targeting in this refractory disease.

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Intratumoral NKp46+ natural killer cells are spatially distanced from T and MHC-I+ cells with prognostic implications in soft tissue sarcoma

Cited by 8 publications

References 51 publications

Spatial colocalization and combined survival benefit of natural killer and CD8 T cells despite profound MHC class I loss in non-small cell lung cancer

Spatial colocalization and combined survival benefit of natural killer and CD8 T cells despite profound MHC class I loss in non-small cell lung cancer

Low-Dose Sorafenib Promotes Cancer Stem Cell Expansion and Accelerated Tumor Progression in Soft Tissue Sarcomas

Characterization of natural killer and cytotoxic T‐cell immune infiltrates in pancreatic ductal adenocarcinoma

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