868 Overall survival on tebentafusp in metastatic uveal melanoma (mUM) across the range of tumor gp100 expression levels

BackgroundTebentafusp is a TCR–anti-CD3 bispecific fusion protein that targets melanoma-expressed gp100 antigen and has shown survival benefit in a randomized phase 3 trial in 1L patients with metastatic uveal melanoma.1 2 In phase 2 and 3 trials (NCT02570308, NCT03070392) enrolling late-stage mUM patients, we explored associations between gp100 expression in the tumor and pharmacodynamic response and clinical outcomes on tebentafusp.Methods2L+ (NCT02570308) or 1L (NCT03070392) HLA-A*02:01+ mUM patients were treated weekly with 68mcg tebentafusp after intra-patient dose escalation. Archival or fresh tumor biopsies were obtained prior to dosing. Expression of baseline gp100 was determined by immunohistochemistry (IHC) and RNAseq analysis (2L+ only) in up to 118 (2L+) and 187 (1L) samples. RNAseq analysis was used to evaluate association between baseline mRNA levels of gp100 and T cell infiltration and activation after 3 doses of tebentafusp (n=35). Serum samples (n=118, 2L+ only) collected at baseline and on-treatment were analyzed for ctDNA. An H-Score quantified tumoral gp100 protein expression. gp100 H score or mRNA levels were cut at the lowest quartile to identify gp100 low patients.ResultsDistribution of gp100 protein by IHC was similar in both studies with median H-Scores of 170 (IQR 60–260) (2L+) and 155 (IQR 68–229) (1L). Over 70% of samples had ≥ 50% gp100+ tumor cells at any intensity. gp100 H-scores were similar in archival and fresh tumor biopsies.High baseline gp100 mRNA levels were associated with ~2-fold increased CD3 and CD8 cell infiltration on tebentafusp compared to little or no change in the gp100 low group. There was greater T cell activation in the gp100 mRNA high group as demonstrated by induction of IFNα (fold change in gp100 high=2.5 p=0.00005,), IFNγ signatures (FC in gp100 high=5.7 p=0.00004) and cytotoxic genes GZMB (FC high=4.6 p=0.000006,) and PRF1 (FC high=2.4 p=0.00051,) compared to little or no activation in the gp100 mRNA low group.Tumor shrinkage (TS) and overall survival (OS) > 12 months were observed in low and high gp100 H-score subgroups (table 1), and a RECIST partial response was observed at very low gp100 (H-score 11). ctDNA reduction on tebentafusp was also observed across the range of gp100 expression levels.Abstract 868 Table 1TS and OS in gp100 high and gp100 low patient groupsConclusionsHigh gp100 expression was associated with the acute pharmacodynamic response to tebentafusp including greater T cell infiltration and activation in the tumor microenvironment. However, clinical outcomes on tebentafusp—TS, OS and ctDNA reduction—were observed across the range of gp100 expression levels.Trial RegistrationNCT02570308, NCT03070392ReferencesMiddleton MR, McAlpine C, Woodcock VK, et al. Tebentafusp, a TCR/Anti-CD3 bispecific fusion protein targeting gp100, potently activated antitumor immune responses in patients with metastatic melanoma. Clin Can Res 2020;26:5869–5878.Sacco JJ, Carvajal R, Butler MO, et al. A phase (ph) II, multi-center study of the safety and efficacy of tebentafusp (tebe) (IMCgp100) in patients (pts) with metastatic uveal melanoma (mUM). Ann Oncol 2020;31:S1442-S1143.Ethics ApprovalThe institutional review board or independent ethics committee at each center approved the trial. The trial was conducted in accordance with the Declaration of Helsinki and the International Conference on Harmonization Good Clinical Practice guidelines.

Download Full-text

Low expression of NSD1, NSD2, and NSD3 define a subset of human papillomavirus-positive oral squamous carcinomas with unfavorable prognosis

Infectious Agents and Cancer ◽

10.1186/s13027-021-00347-6 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Steven F. Gameiro ◽

Farhad Ghasemi ◽

Peter Y. F. Zeng ◽

Neil Mundi ◽

Christopher J. Howlett ◽

...

Keyword(s):

Human Papillomavirus ◽

T Cell ◽

The Cancer Genome Atlas ◽

Cell Infiltration ◽

Relative Level ◽

Expression Levels ◽

T Cell Infiltration ◽

Molecular Features ◽

Platinum Based Chemotherapy ◽

Low Expression

Abstract Background Frequent mutations in the nuclear receptor binding SET domain protein 1 (NSD1) gene have been observed in head and neck squamous cell carcinomas (HNSCC). NSD1 encodes a histone 3 lysine-36 methyltransferase. NSD1 mutations are correlated with improved clinical outcomes and increased sensitivity to platinum-based chemotherapy agents in human papillomavirus-negative (HPV-) tumors, despite weak T-cell infiltration. However, the role of NSD1 and related family members NSD2 and NSD3 in human papillomavirus-positive (HPV+) HNSCC is unclear. Methods Using data from over 500 HNSCC patients from The Cancer Genome Atlas (TCGA), we compared the relative level of mRNA expression of NSD1, NSD2, and NSD3 in HPV+ and HPV- HNSCC. Correlation analyses were performed between T-cell infiltration and the relative level of expression of NSD1, NSD2, and NSD3 mRNA in HPV+ and HPV- HNSCC. In addition, overall survival outcomes were compared for both the HPV+ and HPV- subsets of patients based on stratification by NSD1, NSD2, and NSD3 expression levels. Results Expression levels of NSD1, NSD2 or NSD3 were not correlated with altered lymphocyte infiltration in HPV+ HNSCC. More importantly, low expression of NSD1, NSD2, or NSD3 correlated with significantly reduced overall patient survival in HPV+, but not HPV- HNSCC. Conclusion These results starkly illustrate the contrast in molecular features between HPV+ and HPV- HNSCC tumors and suggest that NSD1, NSD2, and NSD3 expression levels should be further investigated as novel clinical metrics for improved prognostication and patient stratification in HPV+ HNSCC.

Download Full-text

Predicted CD4+ T cell infiltration levels could indicate better overall survival in sarcoma patients

Journal of International Medical Research ◽

10.1177/0300060520981539 ◽

2021 ◽

Vol 49 (1) ◽

pp. 030006052098153

Author(s):

Qing Bi ◽

Yang Liu ◽

Tao Yuan ◽

Huizhen Wang ◽

Bin Li ◽

...

Keyword(s):

Overall Survival ◽

T Cell ◽

Survival Data ◽

Tumor Infiltrating Lymphocytes ◽

The Cancer Genome Atlas ◽

Cell Infiltration ◽

T Cell Infiltration ◽

Whole Exome ◽

Cancer Genome Atlas ◽

Infiltrating Lymphocytes

Objective The role of tumor-infiltrating lymphocytes (TILs) has not yet been characterized in sarcomas. The aim of this bioinformatics study was to explore the effect of TILs on sarcoma survival and genome alterations. Methods Whole-exome sequencing, transcriptome sequencing, and survival data of sarcoma were obtained from The Cancer Genome Atlas. Immune infiltration scores were calculated using the Tumor Immune Estimation Resource. Potential associations between abundance of infiltrating TILs and survival or genome alterations were examined. Results Levels of CD4+ T cell infiltration were associated with overall survival of patients with pan-sarcomas, and higher CD4+ T cell infiltration levels were associated with better survival. Somatic copy number alterations, rather than mutations, were found to correlate with CD4+ T cell infiltration levels. Conclusions This data mining study indicated that CD4+ T cell infiltration levels predicted from RNA sequencing could predict sarcoma prognosis, and higher levels of CD4+ T cells infiltration indicated a better chance of survival.

Download Full-text

342 Combining enadenotucirev and nivolumab increased tumour immune cell infiltration/activation in patients with microsatellite-stable/instability-low metastatic colorectal cancer in a phase 1 study

Journal for ImmunoTherapy of Cancer ◽

10.1136/jitc-2021-sitc2021.342 ◽

2021 ◽

Vol 9 (Suppl 3) ◽

pp. A368-A369

Author(s):

David Krige ◽

Marwan Fakih ◽

Lee Rosen ◽

Ding Wang ◽

Wael Harb ◽

...

Keyword(s):

Colorectal Cancer ◽

T Cell ◽

Metastatic Colorectal Cancer ◽

Cd8 T Cell ◽

Post Treatment ◽

Institutional Review Board ◽

Cell Infiltration ◽

Link Type ◽

T Cell Infiltration ◽

Institutional Review

BackgroundMicrosatellite-stable (MSS) and instability-low (MSI-L) metastatic colorectal cancer (mCRC) are typically characterised as ”immune-excluded/desert” tumour microenvironments lacking T-cell infiltration. Anti-PD-1 monotherapy has little clinical benefit in MSS/MSI-L mCRC1 and knowledge of the effects of PD-1 inhibition on T-cell activation/infiltration in this population is limited. Novel combination therapies to overcome anti-PD-1 resistance are required. SPICE is a multicentre, open-label, phase 1 study of the tumour-selective chimeric Ad11/Ad3 group B oncolytic adenovirus enadenotucirev plus nivolumab in patients with metastatic/advanced epithelial tumours refractory to standard therapy. Preliminary data from patients with MSS/MSI-L mCRC demonstrated a median overall survival of 14 months, manageable tolerability and intratumoural T-cell infiltration.2 Here we characterise the immunological effects of tumour re-engineering with enadenotucirev in combination with nivolumab in patients with MSS/MSI-L mCRC.MethodsPatients received increasing doses and/or cycles of intravenous enadenotucirev followed by up to 8 cycles of nivolumab as previously described.2 Wherever possible, pre- and post-treatment (~5 weeks post-first enadenotucirev) biopsies were collected; samples were analysed using immunohistochemistry and automated image analysis. Peripheral blood mononuclear cell immunophenotyping (multiparameter flow cytometry) and serum cytokines were assessed at multiple times.Results43 patients with mCRC were treated (86% MSS/MSI-L; 14% unknown). Among the 13 patients (12/13 MSS/MSI-L; 1/13 unknown) with matched biopsies, 11 had increased intratumoural and stromal CD8+ T-cell infiltration in post-treatment biopsies (median [Q1-Q3] fold changes 6.5× [1.5–25.4] and 1.9× [1.5–3.9], respectively; figure 1). CD4+ T-cell density increased in 10/13 patients and 8/13 patients had increased proportions of PD-L1+ immune cells. Increases in CD8 T-cell proliferation (Ki67; 7/9 patients) and cytolytic activity (Granzyme B; 7/13 patients) markers were seen. 4/13 patients converted from a ”desert” to an ”inflamed” immune phenotype (pathologist scored CD8/pan-cytokeratin staining). Immunophenotyping showed trends towards increased T-cell activation (CD38+ and HLA-DR+ CD8+ T cell populations) post-treatment (9/10 patients), including in one patient who had only received enadenotucirev prior to sampling. Persistent increases in inflammatory cytokines (IFNγ, IL-12p70, IL-17a) were seen in two patients from ~Day 15, including one who achieved a sustained objective response.Abstract 342 Figure 1Tumour immune cell infiltration following treatment with enadenotucirev plus nivolumabConclusionsThese data show that intravenous enadenotucirev plus nivolumab can induce immune infiltration/activation within MSS/MSI-L mCRC. These encouraging findings suggest that immune activation can be achieved even in ”immune-excluded/desert” tumours. SPICE has been closed following completion of dose-escalation. Efforts are now focused on the development of next-generation variants of enadenotucirev designed to further re-programme the tumour microenvironment by expressing immune-enhancer transgenes (T-SIGn vectors); these studies are ongoing (NCT04830592, NCT04053283, NCT03852511).AcknowledgementsThis study was funded by PsiOxus Therapeutics Limited and Bristol Myers Squibb. Medical writing support: Lola Parfitt, MRes, of PsiOxus Therapeutics Limited.Trial RegistrationEudraCT number2017-001231-39NCT number: NCT02636036ReferencesKawazoe A, Kuboki Y, Shinozaki E, et al. Multicenter phase I/II trial of napabucasin and pembrolizumab in patients with metastatic colorectal cancer (EPOC1503/SCOOP trial). Clin Cancer Res 2020;26:5887–5894.Fakih M, Wang D, Harb W, et al. SPICE: a phase I multicenter study of enadenotucirev in combination with nivolumab in tumors of epithelial origin: an analysis of the metastatic colorectal cancer patients in the dose escalation phase. Ann Oncol 2019:30(suppl_5):v252.Ethics ApprovalThe study was approved by the WCG Institutional Review Board (study approval number 20152656), UCLA Institutional Review Board (study approval number IRB#15-002010), Vanderbilt Institutional Review Board (study approval number IRB #171453) and Henry Ford Institutional Review Board (study approval number IRB #10349).

Download Full-text

Translating transcriptome to immunophenotype in head and neck squamous cell carcinoma (HNSCC) to identify pathways promoting T-cell infiltration.

Journal of Clinical Oncology ◽

10.1200/jco.2019.37.15_suppl.e17542 ◽

2019 ◽

Vol 37 (15_suppl) ◽

pp. e17542-e17542

Author(s):

Theodoros Rampias ◽

Christos K. Kontos ◽

Alexandros Polyzos ◽

Aris Giotakis ◽

Evangelos Giotakis ◽

...

Keyword(s):

T Cell ◽

Checkpoint Inhibitors ◽

Predictive Biomarkers ◽

Keratinocyte Differentiation ◽

Cell Infiltration ◽

Mrna Levels ◽

Rna Seq ◽

Mesenchymal Transition ◽

Invasive Margin ◽

T Cell Infiltration

e17542 Background: We sought to analyze the transcriptional landscape of HNSCC in an attempt to identify tumor-intrinsic oncogenic pathways that appear to mediate T-cell infiltration of tumor tissue. In this direction, we employ a methodology that integrates histopathology data of the tumor microenvironment with its corresponding transcriptome. Methods: 32 frozen HNSCCs were subjected to RNA-seq and corresponding FFPE were scored for plasma cells, tertiary lymphoid structures and CD8a+ TILs (center, invasive margin). RNA-seq data were analyzed to identify differentially expressed genes (DEGs) between tumors scored by immunohistochemistry (IHC) as CD8a high and CD8a low. Gene ontology analysis (GO) was performed based on DEGs > 1.5 fold expression change between CD8a high and CD8a low groups. Candidate genes were investigated by hierarchical clustering in TCGA RNA-seq data and further validated by IHC and quantitative RT-PCR in our cohort. Results: 32 HNSCCs were either scored as CD8a high or CD8a low based on IHC detection of CD8a+ cells in invasive margin of tumors. Comparative analysis of mRNA expression data between CD8a high and CD8a low groups in our cohort revealed that Muc1/16 overexpression and glycosylation was highly enriched in T cell infiltrated group of tumors. This finding was further validated using antibodies that detect glycosylated epitopes for both mucins. Analysis of TCGA RNA-seq data indicated that Muc1/16 overexpressing tumors share signatures of early keratinocyte differentiation and stem cell identity and co-express high levels of enzymes that promote Muc1/16 glycosylation. Interestingly, loss of CDH1 and acquisition of epithelial mesenchymal transition (EMT) markers in the cluster of Muc1/16 overexpressing tumors is strongly correlated with elevated CD8a, IDO1, CD274 and CXCL10 mRNA levels (P < 0.0001). Conclusions: Muc1/16 overexpressing tumors represent a very immunogenic HNSCC cluster. Previous studies have shown that mucins 1 and 16 in cancer cells expose glycosylated-specific epitopes that are recognized by T cells as cancer antigens. To this end, MUC1/16 expression may serve as predictive biomarkers for response to immunotherapy and MUC-targeted immunotherapy may function as an attractive partner to checkpoint inhibitors in HNSCC.

Download Full-text

DPP4 Inhibitor Sitagliptin Enhances Lymphocyte Recruitment and Prolongs Survival in a Syngeneic Ovarian Cancer Mouse Model

Cancers ◽

10.3390/cancers13030487 ◽

2021 ◽

Vol 13 (3) ◽

pp. 487

Author(s):

Amy L. Wilson ◽

Laura R. Moffitt ◽

Kirsty L. Wilson ◽

Maree Bilandzic ◽

Mark D. Wright ◽

...

Keyword(s):

Ovarian Cancer ◽

Overall Survival ◽

T Cell ◽

Mouse Model ◽

Standard Of Care ◽

Cell Infiltration ◽

Metastatic Tumour ◽

Cell Trafficking ◽

T Regulatory Cell ◽

T Cell Infiltration

Immunity plays a key role in epithelial ovarian cancer (EOC) progression with a well-documented correlation between patient survival and high intratumoral CD8+ to T regulatory cell (Treg) ratios. We previously identified dysregulated DPP4 activity in EOCs as a potentially immune-disruptive influence contributing to a reduction in CXCR3-mediated T-cell infiltration in solid tumours. We therefore hypothesized that inhibition of DPP4 activity by sitagliptin, an FDA-approved inhibitor, would improve T-cell infiltration and function in a syngeneic ID8 mouse model of EOC. Daily oral sitagliptin at 50 mg/kg was provided to mice with established primary EOCs. Sitagliptin treatment decreased metastatic tumour burden and significantly increased overall survival and was associated with significant changes to the immune landscape. Sitagliptin increased overall CXCR3-mediated CD8+ T-cell trafficking to the tumour and enhanced the activation and proliferation of CD8+ T-cells in tumour tissue and the peritoneal cavity. Substantial reductions in suppressive cytokines, including CCL2, CCL17, CCL22 and IL-10, were also noted and were associated with reduced CD4+ CD25+ Foxp3+ Treg recruitment in the tumour. Combination therapy with paclitaxel, however, typical of standard-of-care for patients in palliative care, abolished CXCR3-specific T-cell recruitment stimulated by sitagliptin. Our data suggest that sitagliptin may be suitable as an adjunct therapy for patients between chemotherapy cycles as a novel approach to enhance immunity, optimise T-cell-mediated function and improve overall survival.

Download Full-text