Comparison of immune microenvironment between primary lung tumors and paired brain metastatic tumors.

2020 Background: Lung cancer is one of the most common causes of brain metastases (BMs) and is always associated with poor prognosis. To evaluate the characteristics of the tumor immune microenvironment in brain metastases of non-small-cell lung cancer (NSCLC), we investigated the immunophenotype of primary NSCLC and paired brain metastases. Methods: Forty-three Chinese patients with NSCLC who had BMs at presentation or during the course of their disease were admitted to the Sun Yat-Sen University Cancer Center (Guangzhou, China) from 2000 to 2019. RNA sequencing (RNA-seq) of eighty-six formalin-fixed, paraffin embedded (FFPE) samples from primary lung tumors and paired brain metastases of 43 patients was conducted to comprehensively analyze the tumor immune microenvironment. Results: Our data revealed that brain metastases compared with primary lung tumors exhibited reduced tumor infiltrating lymphocytes (TILs) (all 28 immune cell subtypes P < 0.05), lower fraction of activated CD8 T cell and effector memory CD8 T cell in total TILs (P = 0.028, P < 0.001, respectively); higher fraction of macrophage and neutrophil in total TILs (P < 0.001, P < 0.01, respectively). Comparing with the primary lung tumors, the scores of some immune related signatures, including MHC non-class signature, IFN gamma signature and T-cell-inflamed gene-expression profile (GEP) signature, were significantly lower in brain metastases (P = 0.004, P = 0.009, P = 0.004, respectively), while the score of MHC class-II signature was higher in brain metastases (P = 0.045). We found the distributions of tumor microenvironment immune types (TMIT) in brain metastases and primary lung tumors were different. Brain metastases contained significantly lower proportion of TMIT I (high PD-L1/ high CD8A) (23%) than primary lung tumors (47%) (P < 0.05). Besides, we found three immune inhibitory checkpoint molecules, namely C10orf54 (VISTA), CTLA4 and CD274 (PD-L1) were downregulated in brain metastases than in primary lung tumors (P < 0.001, P < 0.001, P = 0.034, respectively). Moreover, there was poor correlation of PD-L1 expression between paired brain metastases and primary lung tumors (R = 0.28, P = 0.068). Unsupervised hierarchic cluster analysis revealed the primary lung tumors had two distinct patterns of immune gene signatures, namely Cluster A and Cluster B, and the tumors in Cluster B were immune rich, but associated with poor prognosis (log-rank P = 0.021). Conclusions: Our work illustrates the immune landscape of brain metastases from NSCLC, and suggests that the tumor immune microenvironment in brain metastases compared with primary lung tumors is further immunosuppressed, that may help guide immunotherapeutic strategies for NSCLC brain metastases.

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Functional profile and clinical significance of major tumor infiltrating lymphocyte subsets in lung cancer-associated brain metastases.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.2066 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. 2066-2066

Author(s):

Benjamin Y. Lu ◽

Richa Gupta ◽

Tyler Stewart ◽

Harriet M. Kluger ◽

Lucia Jilaveanu ◽

...

Keyword(s):

Lung Cancer ◽

T Cells ◽

T Cell ◽

Granzyme B ◽

Lung Tumors ◽

Small Cell ◽

T Cell Subsets ◽

Functional Profile ◽

Ki 67 ◽

Primary Lung Tumors

2066 Background: Despite the biological and clinical implications, the immune composition and functional characteristics of adaptive immune cells in brain metastases (BrM) are poorly understood. Using multiplexed quantitative immunofluorescence (QIF), this study evaluates the level and functional profile of major T-cell subsets in primary lung tumors, BrM, and extracranial metastases (ECM) from lung cancers. Methods: A tissue microarray was constructed from formalin-fixed, paraffin-embedded tumor samples of 94 lung cancer patients treated at Yale Cancer Center between 2002-2013. Multiplexed QIF was used to evaluate the cases with a panel containing phenotype markers for major T-cell subsets (CD3, CD4, CD8 and FOXP3), and cell-localized activation and proliferation (granzyme-B and Ki-67). Signal for each marker was measured in marker-selected tissue compartments using the Automated Quantitative Analysis (AQUA) platform. Associations between markers and major clinicopathologic variables were studied. Results: In total, 40 primary lung tumors, 63 BrM, and 15 ECM were analyzed, including paired samples from 22 patients. Lung cancer histology included adenocarcinoma 62.5%, squamous cell carcinoma 11.5%, small cell 9.4%, and other non-small cell 16.7%. BrM had both significantly lower levels of CD3+ T-cells ( p< 0.0001) and T-cell granzyme B ( p= 0.0188) compared with primary lung tumors. No significant differences were observed in T-cell Ki-67 levels across tissues. FOXP3 measured in CD4+ T-cells were significantly lower in BrM compared with primary malignancies ( p= 0.0002) and ECM ( p= 0.0404). Among patients with BrM, higher levels of CD3+ T-cells in BrM were associated with longer overall survival. Conclusions: Lung cancer-associated BrM have lower T-cell infiltration, cytolytic function, and regulatory T-cells than primary lesions. These results indicate lower adaptive anti-tumor responses in BrM and suggest the presence of a tolerogenic microenvironment in the brain. Overcoming this could be used to design optimal treatment strategies.

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303 Non-functional T cell responses in non-small cell lung cancer are induced during tumor antigen-specific T cell priming in the mediastinal lymph node

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2021-sitc2021.303 ◽

2021 ◽

Vol 9 (Suppl 3) ◽

pp. A326-A326

Author(s):

Brendan Horton ◽

Duncan Morgan ◽

Elen Torres-Mejia ◽

Maria Zagorulya ◽

Vidit Bhandarkar ◽

...

Keyword(s):

Lung Cancer ◽

T Cells ◽

T Cell ◽

Cd8 T Cells ◽

Lung Tumor ◽

T Cell Responses ◽

Cd8 T Cell ◽

Lung Tumors ◽

Tumor Control ◽

Cell Responses

BackgroundIn non-small cell lung cancer (NSCLC), response to checkpoint blockade therapy (CBT) is associated with tumor-infiltrating CD8+ T cells, but not all T cell-infiltrated tumors respond to CBT. The subgroup of T cell-infiltrated but CBT-resistant tumors has been clinically described as containing ”non-functional” T cell responses. Mechanisms governing the generation of non-functional T cell responses remain poorly understood, and treatment options for this subgroup are limited.MethodsWe utilized a transplantable, syngeneic murine NSCLC cell line derived from an autochthonous NSCLC driven by KrasG12D expression and p53 deletion (KP cell line) to model non-functional T cell responses. To study antigen-specific responses, we engineered KP cells to express the model CD8+ T cell antigen SIY for certain experiments. CBT consisted of combined anti-CTLA-4 and anti-PD-L1 therapy.ResultsOrthotopic KP lung tumors failed to respond to CBT, but KP flank tumors were controlled by CBT. Lung and flank tumors contained activated CD8+ T cells, providing a platform to compare functional and non-functional CD8+ T cell responses in NSCLC. Single-cell RNA sequencing revealed that lung tumor-infiltrating CD8+ T cells lacked effector and exhaustion molecules despite clonal expansion. Analysis of antigen-specific CD8+ T cells revealed that this lung cancer-specific T cell dysfunction was established during priming in lung-draining mediastinal lymph nodes (mLN) despite robust T cell proliferation. RNA sequencing and flow cytometry of antigen-specific CD8+ T cells found that T cells primed in the mLN underwent blunted effector differentiation characterized by a lack of effector molecules CD25, Granzyme B, and TIM-3, but retention of TCF-1. This phenotype persisted in lung tumors, consistent with our initial observations of absent effector and exhaustion molecule expression. Many CD8+ T cells from NSCLC patients expressed an analogous gene expression program distinct from T cell exhaustion. TCF-1+ CD8+ T cells in lung tumors did not mediate tumor control and failed to differentiate into effector cells after CBT. To investigate alternative therapeutic strategies of reinvigorating lung tumor-reactive T cells, we focused on IL-2 and IL-12, as expression of their receptors was reduced in mLN-primed T cells. Administering recombinant IL-2 and IL-12 was sufficient to restore effector T cell differentiation, induce lung tumor control, and significantly extend survival of lung tumor-bearing mice.ConclusionsOur results suggest that non-functional CD8+ T cell responses in NSCLC arise from failed effector T cell differentiation during priming. Transient combination therapy with IL-2 and IL-12 overcomes this dysfunctional state to induce protective T cell responses in CBT-resistant tumors.Ethics ApprovalAll mouse experiments were approved by MIT’s Committee on Animal Care (CAC) - DHHS Animal Welfare Assurance # D16-00078

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Tumor-Induced CD8+ T-Cell Dysfunction in Lung Cancer Patients

Clinical and Developmental Immunology ◽

10.1155/2012/741741 ◽

2012 ◽

Vol 2012 ◽

pp. 1-11 ◽

Cited By ~ 42

Author(s):

Heriberto Prado-Garcia ◽

Susana Romero-Garcia ◽

Dolores Aguilar-Cazares ◽

Manuel Meneses-Flores ◽

Jose Sullivan Lopez-Gonzalez

Keyword(s):

Lung Cancer ◽

T Cells ◽

T Cell ◽

Cd8 T Cells ◽

Cancer Biology ◽

Lung Tumor ◽

Tumor Infiltrating Lymphocytes ◽

Cd8 T Cell ◽

Lung Tumors ◽

Autologous Tumor

Lung cancer is the leading cause of cancer deaths worldwide and one of the most common types of cancers. The limited success of chemotherapy and radiotherapy regimes have highlighted the need to develop new therapies like antitumor immunotherapy. CD8+ T-cells represent a major arm of the cell-mediated anti-tumor response and a promising target for developing T-cell-based immunotherapies against lung cancer. Lung tumors, however, have been considered to possess poor immunogenicity; even so, lung tumor-specific CD8+ T-cell clones can be established that possess cytotoxicity against autologous tumor cells. This paper will focus on the alterations induced in CD8+ T-cells by lung cancer. Although memory CD8+ T-cells infiltrate lung tumors, in both tumor-infiltrating lymphocytes (TILs) and malignant pleural effusions, these cells are dysfunctional and the effector subset is reduced. We propose that chronic presence of lung tumors induces dysfunctions in CD8+ T-cells and sensitizes them to activation-induced cell death, which may be associated with the poor clinical responses observed in immunotherapeutic trials. Getting a deeper knowledge of the evasion mechanisms lung cancer induce in CD8+ T-cells should lead to further understanding of lung cancer biology, overcome tumor evasion mechanisms, and design improved immunotherapeutic treatments for lung cancer.

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An Immune Gene-Related Five-lncRNA Signature for to Predict Glioma Prognosis

Frontiers in Genetics ◽

10.3389/fgene.2020.612037 ◽

2020 ◽

Vol 11 ◽

Author(s):

Xinzhuang Wang ◽

Ming Gao ◽

Junyi Ye ◽

Qiuyi Jiang ◽

Quan Yang ◽

...

Keyword(s):

T Cell ◽

Risk Scores ◽

Normal Brain ◽

Immune Gene ◽

Immune Microenvironment ◽

Risk Models ◽

Immune State ◽

M1 Macrophage ◽

Tumor Immune Microenvironment ◽

Patient Prognosis

BackgroundThe tumor immune microenvironment is closely related to the malignant progression and treatment resistance of glioma. Long non-coding RNA (lncRNA) plays a regulatory role in this process. We investigated the pathological mechanisms within the glioma microenvironment and potential immunotherapy resistance related to lncRNAs.MethodWe downloaded datasets derived from glioma patients and analyzed them by hierarchical clustering. Next, we analyzed the immune microenvironment of glioma, related gene expression, and patient survival. Coexpressed lncRNAs were analyzed to generate a model of lncRNAs and immune-related genes. We analyzed the model using survival and Cox regression. Then, univariate, multivariate, receiver operating characteristic (ROC), and principle component analysis (PCA) methods were used to verify the accuracy of the model. Finally, GSEA was used to evaluate which functions and pathways were associated with the differential genes.ResultsNormal brain tissue maintains a low-medium immune state, and gliomas are clearly divided into three groups (low to high immunity). The stromal, immune, and estimate scores increased along with immunity, while tumor purity decreased. Further, human leukocyte antigen (HLA), programmed cell death-1 (PDL1), T cell immunoglobulin and mucin domain 3 (TIM-3), B7-H3, and cytotoxic T lymphocyte-associated antigen-4 (CTLA4) expression increases concomitantly with immune state, and the patient prognosis worsens. Five immune gene-related lncRNAs (AP001007.1, LBX-AS1, MIR155HG, MAPT-AS1, and LINC00515) were screened to construct risk models. We found that risk scores are related to patient prognosis and clinical characteristics, and are positively correlated with PDL1, TIM-3, and B7-H3 expression. These lncRNAs may regulate the tumor immune microenvironment through cytokine–cytokine receptor interactions, complement, and coagulation cascades, and may promote CD8 + T cell, regulatory T cell, M1 macrophage, and infiltrating neutrophils activity in the high-immunity group. In vitro, the abnormal expression of immune-related lncRNAs and the relationship between risk scores and immune-related indicators (PDL1, CTLA4, CD3, CD8, iNOS) were verified by q-PCR and immunohistochemistry (IHC).ConclusionFor the first time, we constructed immune gene-related lncRNA risk models. The risk score may be a new biomarker for tumor immune subtypes and provide molecular targets for glioma immunotherapy.

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Integrative and Comparative Genomic Analysis and the Immune Microenvironment Features of Lung Cancer Patients with Tuberculosis

10.21203/rs.3.rs-667647/v1 ◽

2021 ◽

Author(s):

Xiaoling Xu ◽

Chaohui Bao ◽

Da Chen ◽

Tianxiang Wang ◽

Changchun Wang ◽

...

Keyword(s):

Lung Cancer ◽

T Cell ◽

Cd8 T Cell ◽

Tumour Microenvironment ◽

Cell Infiltration ◽

Driver Mutations ◽

Comparative Genomic ◽

Immune Microenvironment ◽

Genomic Alterations ◽

T Cell Infiltration

Abstract Background: Limited information was known because of the low incidence of co-existence with lung cancer and tuberculosis (TB), it remains special challenging populations for clinical management of cancer immunotherapy. Thus, to investigate the difference on tumour immune microenvironment and genomics between patients with LC alone and LC patients with TB is urgently needed. Methods:Tumour specimens were collected from 87 patients who had LC, with or without TB, at two medical centres. Immunohistochemistry was used to evaluate PD-L1 expression and CD3+/CD4+/CD8+ T-cell infiltration. Whole-exome sequencing was performed using samples from 19 patients with LC&TB and 21 patients with LC. Results:Relative to patients with LC alone, patients with LC&TB had lower PD-L1 expression and CD4+/CD8+ T-cell infiltration (all P< 0.001). A tumour microenvironment with no PD-L1 expression and CD8- T-cell infiltration was most common in the LC&TB patients. Genomic alterations analysis revealed an increased mutation frequency among patients with LC and active TB, obsolete/cured TB, or no TB in terms of the TP53 (23.08% vs. 66.67% vs. 76.19%, P = 0.01), while a decreased trend of the number of single-nucleotide variants/insertions/deletions (P< 0.001), tumour mutation burden (P< 0.001), and number of neoantigens (P< 0.001). Patients with LC&TB had a higher frequency of a specific mutation signature (32.99% vs. 11.23%), as well as potential driver mutations involving the complement C1qB chain (C1QB) mutations. Conclusion: The present study revealed significant differences in the tumour microenvironment and genomic alterations between patients with LC&TB and patients with LC alone.

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Genomic profiles and tumor immune microenvironment of primary lung carcinoma and brain oligo-metastasis

Cell Death and Disease ◽

10.1038/s41419-021-03410-7 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Zhengbo Song ◽

Ling Yang ◽

Zhipeng Zhou ◽

Pansong Li ◽

Wenxian Wang ◽

...

Keyword(s):

Lung Cancer ◽

T Cell ◽

Brain Metastasis ◽

Lung Carcinoma ◽

Primary Tumor ◽

Lung Tumor ◽

Distinct Pattern ◽

Cytotoxic T Cell ◽

Immune Microenvironment ◽

Tumor Immune Microenvironment

AbstractBrain metastasis (BM) is a common malignant event in lung cancer. Here, we recruited 33 lung cancer patients with brain oligo-metastasis to explore the genomic features and tumor immune microenvironment (TIME) of the lung and BM independently. For genomic profiling, targeted sequencing was performed. We found that high-frequent ZFHX3 occurred in the lung (40%) and brain tumor (28%), which might relate to brain metastasis event; the vast majority of patients had lesions-shared mutations in primary tumor and BM, confirming the common clonal events; and EGFR was the most frequently clonal gene in both lung and BM, indicating its driver capability. To characterize TIME status, we also sequenced the T cell receptor (TCR) repertoires and performed immunohistochemistry (IHC) on CD8+ tumor-infiltrating lymphocytes (TILs) and PD-L1 expression in 28 patients who had paired samples. Through the comparison, the TCR clonality of BM was higher than lung tumor, indicating the distinct pattern of the stronger oligoclonal T cell expansion in BM; the primary tumor had a higher TMB than oligo-BM (13.9 vs 8.7 mutations, p = 0.019); CD8 + TILs of BM were significantly lower than lung tumor (10% vs 30%, p = 0.015), revealing the lower level of cytotoxic T cell infiltration; BM showed statistically equivalent level of PD-L1 compared with lung tumor (p = 0.722). We further investigated the potential biomarkers associated with overall survival (OS) after brain surgery. We found that higher TCR clonality was related to prolonged OS in EGFR-treated patients (HR 0.175, p < 0.001) but the worse outcomes in non-EGFR-treated (HR 2.623, p = 0.034). More CD8+ TILs were an independently positive indicator for OS, in EGFR-treated (HR 0.160, p = 0.001) and non-EGFR-treated patients (HR 0.308, p = 0.009). These findings provide a meaningful molecular and clinical understanding of lung carcinoma and brain oligo-metastasis.

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BSCI-09. Multiomic single cell analysis reveals emerging principles of tumor immune microenvironment inherent to NSCLC brain metastases

Neuro-Oncology Advances ◽

10.1093/noajnl/vdab071.008 ◽

2021 ◽

Vol 3 (Supplement_3) ◽

pp. iii3-iii3

Author(s):

Lei Wen ◽

Changguo Shan ◽

Da Liu ◽

Cheng Zhou ◽

Linbo Cai

Keyword(s):

Lung Cancer ◽

Brain Metastasis ◽

Brain Metastases ◽

Single Cell ◽

Cancer Patients ◽

Cancer Cell ◽

Immune Microenvironment ◽

Lung Cancer Patients ◽

Tumor Immune Microenvironment ◽

Treatment Naïve

Abstract Brain is one of the most common sites for distant metastasis of lung cancer. Treatment naïve lung cancer patients diagnosed with brain metastasis are left with very limited options. Checkpoint inhibition is a powerful immunotherapy strategy but delivers benefit only to a small population of patients. Here we harnessed the power and resolution of single cell RNA sequencing and single cell TCR/BCR sequencing to investigate the tumor immune microenvironment (TIME) of NSCLC brain metastases. We enrolled treatment naïve lung cancer patients with brain metastasis. The enrolled subjects covered different histology types and driver gene mutation status. We revealed the emerging principles of innate and adaptive immune components inherent to NSCLC brain metastases. We also uncovered several significant intercellular communication patterns that potentiates cancer cell seeding and fosters cancer cell proliferation. Those results served as a starting point to design optimal immunotherapy strategies for advanced lung cancer patients with limited options.

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