The mutational load and a T-cell inflamed tumour phenotype identify ovarian cancer patients rendering tumour-reactive T cells from PD-1+ tumour-infiltrating lymphocytes

Abstract Background Adoptive immunotherapy with tumour-infiltrating lymphocytes (TIL) may benefit from the use of selective markers, such as PD-1, for tumour-specific T-cell enrichment, and the identification of predictive factors that help identify those patients capable of rendering tumour-reactive TILs. We have investigated this in ovarian cancer (OC) patients as candidates for TIL therapy implementation. Methods PD-1− and PD-1+ CD8 TILs were isolated from ovarian tumours and expanded cells were tested against autologous tumour cells. Baseline tumour samples were examined using flow cytometry, multiplexed immunofluorescence and Nanostring technology, for gene expression analyses, as well as a next-generation sequencing gene panel, for tumour mutational burden (TMB) calculation. Results Tumour-reactive TILs were detected in half of patients and were exclusively present in cells derived from the PD-1+ fraction. Importantly, a high TIL density in the fresh tumour, the presence of CD137+ cells within the PD-1+CD8+ TIL subset and their location in the tumour epithelium, together with a baseline T-cell-inflamed genetic signature and/or a high TMB, are features that identify patients rendering tumour-reactive TIL products. Conclusion We have demonstrated that PD-1 identifies ovarian tumour-specific CD8 TILs and has uncovered predictive factors that identify OC patients who are likely to render tumour-specific cells from PD-1+ TILs.

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The mutational load and a T-cell inflamed tumor phenotype identify ovarian cancer patients rendering tumor-reactive T cells from PD-1+ tumor-infiltrating lymphocytes

10.21203/rs.3.rs-25670/v1 ◽

2020 ◽

Cited By ~ 1

Author(s):

Diego Salas-Benito ◽

Enrique Conde ◽

Ibon Tamayo-Uria ◽

Uxua Mancheño ◽

Edurne Elizalde ◽

...

Keyword(s):

Ovarian Cancer ◽

T Cells ◽

T Cell ◽

Predictive Biomarkers ◽

Tumor Infiltrating Lymphocytes ◽

Ovarian Tumors ◽

Autologous Tumor ◽

Tumor Mutational Burden ◽

Nanostring Technology ◽

Infiltrating Lymphocytes

Abstract Background: Adoptive immunotherapy with tumor-infiltrating lymphocytes (TIL) may benefit from the use of selective markers, such as programmed cell death protein 1 (PD-1), for tumor-specific T-cell enrichment, as well as predictive biomarkers that help identify those patients capable of rendering tumor-reactive TIL products. We have investigated this in ovarian cancer (OC) patients as candidate for TIL therapy implementation. Methods: PD-1- and PD-1+ CD8 TILs were isolated from resected ovarian tumors and, after polyclonal expansion, TIL products were tested against autologous tumor cells. Reactivity was assessed by IFNg production (ELISPOT) and upregulation of CD137. Baseline tumor samples were examined using flow cytometry, multiplexed quantitative immunofluorescence, Nanostring technology, for gene expression profile (GEP) analyses, as well as a next generation sequencing gene panel, for tumor mutational burden (TMB) calculation, to identify those features that distinguished patients with tumor-reactive and non-tumor-reactive TIL products.Results: Tumor-reactive TILs were detected in half of patients and were exclusively present in cells derived from the PD-1+ fraction. Flow-cytometric studies revealed that fresh tumors from patients rendering tumor-reactive TILs presented a significantly higher frequency of CD137+ cells within the PD1+CD8+ subset. Multiplexed immunofluorescence supported this finding, which was particularly striking in intraepithelial CD8 TILs. Baseline GEP analysis showed that patients rendering tumor-reactive TILs exhibited a significantly higher T-cell inflamed signature. Despite no correlation between TMB and GEP, both parameters stratified tumors, with patients with higher TMB and/or T-cell inflamed signature score rendering tumor-reactive TILs. Conclusion: We have demonstrated that PD-1 identifies autologous-tumor specific CD8 T cells infiltrating ovarian tumors and have uncovered predictive factors that identify OC patients who are likely to render tumor-reactive cells from PD-1+ TILs. These findings have important implications for improving the efficacy of TIL therapy in OC.

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High-throughput sequencing of T-cell receptors reveals a homogeneous repertoire of tumour-infiltrating lymphocytes in ovarian cancer

The Journal of Pathology ◽

10.1002/path.4260 ◽

2013 ◽

Vol 231 (4) ◽

pp. 433-440 ◽

Cited By ~ 98

Author(s):

Ryan O Emerson ◽

Anna M Sherwood ◽

Mark J Rieder ◽

Jamie Guenthoer ◽

David W Williamson ◽

...

Keyword(s):

Ovarian Cancer ◽

T Cell ◽

High Throughput ◽

T Cell Receptors ◽

High Throughput Sequencing ◽

Cell Receptors ◽

Tumour Infiltrating Lymphocytes ◽

Infiltrating Lymphocytes

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Cytotoxic T-Cell Clones Isolated from Ovarian Tumour Infiltrating Lymphocytes Recognize Common Determinants on Non-Ovarian Tumour Clones

Scandinavian Journal of Immunology ◽

10.1111/j.1365-3083.1993.tb03312.x ◽

1993 ◽

Vol 37 (4) ◽

pp. 413-424 ◽

Cited By ~ 19

Author(s):

C. G. IOANNIDES ◽

B. FISK ◽

M. S. POLLACK ◽

M. L. FRAZIER ◽

J. TAYLOR WHARTON ◽

...

Keyword(s):

T Cell ◽

Ovarian Tumour ◽

Cytotoxic T Cell ◽

T Cell Clones ◽

Cell Clones ◽

Tumour Infiltrating Lymphocytes ◽

Infiltrating Lymphocytes

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Characterization of fresh (uncultured) tumour-infiltrating lymphocytes (TIL) and TIL-derived T cell lines from patients with renal cell carcinoma

Clinical & Experimental Immunology ◽

10.1111/j.1365-2249.1994.tb06088.x ◽

2008 ◽

Vol 97 (2) ◽

pp. 321-327 ◽

Cited By ~ 19

Author(s):

D. MITROPOULOS ◽

S. KOOI ◽

J. RODRIGUEZ-VILLANUEVA ◽

C. D. PLATSOUCAS

Keyword(s):

Renal Cell Carcinoma ◽

T Cell ◽

Cell Carcinoma ◽

Cell Lines ◽

Renal Cell ◽

Tumour Infiltrating Lymphocytes ◽

T Cell Lines ◽

Infiltrating Lymphocytes

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Engineering adoptive T cell therapy to co-opt Fas ligand-mediated death signaling in ovarian cancer enhances therapeutic efficacy

10.1101/2021.07.30.454539 ◽

2021 ◽

Author(s):

Kristin G. Anderson ◽

Shannon K. Oda ◽

Breanna M. Bates ◽

Madison G. Burnett ◽

Magdalia Rodgers Suarez ◽

...

Keyword(s):

Ovarian Cancer ◽

T Cells ◽

Cell Death ◽

T Cell ◽

Cell Therapy ◽

Tumor Vasculature ◽

Adoptive T Cell Therapy ◽

T Cell Therapy ◽

Tumor Bearing ◽

Infiltrating Lymphocytes

Background: In the U.S., more than 50% of ovarian cancer patients die within 5 years of diagnosis, highlighting the need for innovations such as engineered T cell therapies. Mesothelin (Msln) is an attractive immunotherapy target for this cancer, as it is overexpressed by the tumor and contributes to malignant and invasive phenotypes, making antigen loss disadvantageous to the tumor. We previously showed that adoptively transferred T cells engineered to be Msln-specific (TCR1045) preferentially accumulate within established ovarian tumors, delay tumor growth and significantly prolong survival in the ID8VEGF mouse model. However, T cell persistence and anti-tumor activity were not sustained, and we and others have previously detected FasL in the tumor vasculature and the tumor microenvironment (TME) of human and murine ovarian cancers, which can induce apoptosis in infiltrating lymphocytes expressing Fas receptor (Fas). Methods: To concurrently overcome this mechanism for potential immune evasion and enhance T cell responses, we generated an immunomodulatory fusion protein (IFP) containing the Fas extracellular binding domain fused to a 4-1BB co-stimulatory domain, rather than the natural death domain. T cells engineered to express TCR1045 alone or in combination with the IFP were transferred into ID8VEGF-tumor bearing mice and evaluated for persistence, proliferation, anti-tumor cytokine production, and therapeutic efficacy. Results: Relative to T cells modified only to express TCR1045, T cells engineered to express both TCR1045 and a Fas IFP preferentially persisted in the TME of tumor-bearing mice due to improved T cell proliferation and survival. Moreover, adoptive immunotherapy with IFP+ T cells significantly prolonged survival in tumor-bearing mice, relative to TCR1045 T cells lacking the IFP. Conclusions: Fas/FasL signaling can mediate T cell death in the ovarian cancer microenvironment, as well as induce activation-induced cell death, an apoptotic mechanism responsible for regulating T cell expansion. Upregulation of FasL by tumor cells and tumor vasculature represents a mechanism for protecting growing tumors from attack by tumor-infiltrating lymphocytes. As many solid tumors overexpress FasL, an IFP that converts the Fas-mediated death signal into pro-survival and proliferative signals may provide an opportunity to enhance engineered adoptive T cell therapy against many malignancies.

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Analysis of T-Cell Receptor Variability in Tumour Infiltrating Lymphocytes

Lymphohaematopoietic Growth Factors in Cancer Therapy II ◽

10.1007/978-3-642-77801-8_6 ◽

1992 ◽

pp. 49-55

Author(s):

Frédéric Triebel

Keyword(s):

T Cell ◽

T Cell Receptor ◽

Cell Receptor ◽

Tumour Infiltrating Lymphocytes ◽

Infiltrating Lymphocytes

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Characterization of T cell repertoire of blood, tumor, and ascites in ovarian cancer patients using next generation sequencing

OncoImmunology ◽

10.1080/2162402x.2015.1030561 ◽

2015 ◽

Vol 4 (11) ◽

pp. e1030561 ◽

Cited By ~ 34

Author(s):

Miran Jang ◽

Poh-Yin Yew ◽

Kosei Hasegawa ◽

Yuji Ikeda ◽

Keiichi Fujiwara ◽

...

Keyword(s):

Ovarian Cancer ◽

Next Generation Sequencing ◽

T Cell ◽

Cancer Patients ◽

T Cell Repertoire ◽

Next Generation ◽

Cell Repertoire ◽

Generation Sequencing

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Qualitative Analysis of Tumor-Infiltrating Lymphocytes across Human Tumor Types Reveals a Higher Proportion of Bystander CD8+ T Cells in Non-Melanoma Cancers Compared to Melanoma

Cancers ◽

10.3390/cancers12113344 ◽

2020 ◽

Vol 12 (11) ◽

pp. 3344

Author(s):

Aishwarya Gokuldass ◽

Arianna Draghi ◽

Krisztian Papp ◽

Troels Holz Borch ◽

Morten Nielsen ◽

...

Keyword(s):

Ovarian Cancer ◽

T Cells ◽

T Cell ◽

Tumor Infiltrating Lymphocytes ◽

Autologous Tumor ◽

Epithelial Cancers ◽

Single Cell Rna Sequencing ◽

Tumor Types ◽

Infiltrating Lymphocytes

Background: Human intratumoral T cell infiltrates can be defined by quantitative or qualitative features, such as their ability to recognize autologous tumor antigens. In this study, we reproduced the tumor-T cell interactions of individual patients to determine and compared the qualitative characteristics of intratumoral T cell infiltrates across multiple tumor types. Methods: We employed 187 pairs of unselected tumor-infiltrating lymphocytes (TILs) and autologous tumor cells from patients with melanoma, renal-, ovarian-cancer or sarcoma, and single-cell RNA sequencing data from a pooled cohort of 93 patients with melanoma or epithelial cancers. Measures of TIL quality including the proportion of tumor-reactive CD8+ and CD4+ TILs, and TIL response polyfunctionality were determined. Results: Tumor-specific CD8+ and CD4+ TIL responses were detected in over half of the patients in vitro, and greater CD8+ TIL responses were observed in melanoma, regardless of previous anti-PD-1 treatment, compared to renal cancer, ovarian cancer and sarcoma. The proportion of tumor-reactive CD4+ TILs was on average lower and the differences less pronounced across tumor types. Overall, the proportion of tumor-reactive TILs in vitro was remarkably low, implying a high fraction of TILs to be bystanders, and highly variable within the same tumor type. In situ analyses, based on eight single-cell RNA-sequencing datasets encompassing melanoma and five epithelial cancers types, corroborated the results obtained in vitro. Strikingly, no strong correlation between the proportion of CD8+ and CD4+ tumor-reactive TILs was detected, suggesting the accumulation of these responses in the tumor microenvironment to follow non-overlapping biological pathways. Additionally, no strong correlation between TIL responses and tumor mutational burden (TMB) in melanoma was observed, indicating that TMB was not a major driving force of response. No substantial differences in polyfunctionality across tumor types were observed. Conclusions: These analyses shed light on the functional features defining the quality of TIL infiltrates in cancer. A significant proportion of TILs across tumor types, especially non-melanoma, are bystander T cells. These results highlight the need to develop strategies focused on the tumor-reactive TIL subpopulation.

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