Common phenotypic dynamics of tumor-infiltrating lymphocytes across different histologies upon checkpoint inhibition: impact on clinical outcome

doi:10.1016/j.jcyt.2020.01.010

Cytotherapy

Volume 22, Issue 4, April 2020, Pages 204-213

https://doi.org/10.1016/j.jcyt.2020.01.010 Get rights and content

Abstract

Background

Immune checkpoint inhibitors (ICIs) have revolutionized the cancer therapeutic landscape and our perception of interactions between the immune system and tumor cells. Despite remarkable progress, disease relapse and primary resistance are not uncommon. Understanding the biological processes that tumor-infiltrating lymphocytes (TILs) undergo during ICI, how this affects the tumor microenvironment (TME) and, ultimately, clinical outcome is, therefore, necessary to further improve treatment efficacy.

Aim

In the current study, we sought to characterize TILs from patients with metastatic solid tumors undergoing ICI correlating flowcytometric findings with clinical outcome.

Methods

In total, 20 patients with 10 different metastatic solid tumors treated with ICIs targeting programmed-cell death-1 (PD-1)/PD-L1 axis were included in this study. The phenotype of T cells deriving from biopsies obtained prior to treatment initiation and on-treatment was investigaded. Analyses were focused on T cells’ degree of differentiation and activity and how they correlate with transcriptomic changes in the TME.

Results

Data indicate that patients benefitting from ICIs accumulate CD8+central memory T cells. TILs developed an effector-like phenotype over time, which was also associated with a cytolytic gene signature. In terms of modulation of T-cell responses, we observed that high expression of checkpoint molecules pre-treatment (i.e., PD-1, lymphocyte activation gene-3 [LAG-3], B and T-lymphocyte attenuator [BTLA] and T-cell immunoglobulin and mucin domain containing-3 [TIM-3]) was associated with similar gene signature and correlated to treatment benefit. Increasing expression of LAG-3 and BTLA in the CD8 compartment and their co-expression with PD-1 during treatment were, however, a common feature for patients who failed to respond to ICIs.

Conclusions

Besides identifying immune profiles suggestive of response to ICI, our results provide a more nuanced picture regarding expression of checkpoint molecules that goes beyond T-cell anergy.

Graphical Abstract

Section snippets

Background

The presence of immune cells in the tumor microenvironment (TME) has been long known [1], but it is only over the past decades that targeting the immune system as treatment strategy against cancer has gained popularity, especially due to remarkable results yielded by immune checkpoint inhibitors (ICIs) [2], [3], [4].

The rationale behind ICIs is to revert the state of T-cell dysfunctionality where continuous TCR stimulation by chronic antigen exposure in an immune suppressive milieu has led to

Study design

We designed a basket study to investigate shared T-cell phenotypic features and their correlation to genetic changes across different tumor types. The presented study has, therefore, an explorative character and hypothesis-generating intent.

The protocol related to the study was approved by The Danish National Committee on Health Research Ethics (H-16046968) and the Danish Data Protection Agency (RH-2018-44). Written informed consent was obtained prior to any collection and storage of biological

Patients characteristics

From February 2017 to December 2018, 37 patients referred to our service for treatment with ICIs were screened for inclusion in this study. In total, eight patients were excluded due to withdrawal of informed consent (n = 3) and change of therapeutic strategy after first visit (n = 5). The remaining 29 patients were referred for radiologically guided tumor biopsies. The screening process and number of samples used in the study are illustrated in the consort diagram (Supplementary Figure 2). The

Discussion

Despite the remarkable response rates yielded by ICI in cohorts of melanoma [2,27] and non–small cell lung cancer [28], a considerable fraction of patients still do not respond to the therapy [5]. A better understanding of such variable outcomes requires insight into the biological basis for treatment success and failure.

In our study, we explored the functional repertoire of TILs isolated from patients undergoing ICI, prior to treatment start and on-treatment, searching for common elements,

Funding

This study was only possible due to financial support granted by The Danish Cancer Society and Preben & Anna Simonsens Fond.

Disclosure of Competing Interest

The authors have no commercial, proprietary, or financial interest in the products or companies described in this article.

Author Contributions

VL was responsible for conception and design of the study, idea development, inclusion of patients, collection of biological material and clinical data, flow cytometry acquiring/gating, statistical analysis and data visualization, manuscript writing and final approval of the manuscript. AB was responsible for bioinformatic analyses, interpretation of data, critical review and final approval of the manuscript. MH was responsible for assistance with flow cytometry data, critical review and final

References (52)

S.W. Lim et al.
Current status of immune checkpoint inhibitors in treatment of non-small cell lung cancer
Korean J Intern Med
(2019)
M.S. Rooney et al.
Molecular and genetic properties of tumors associated with local immune cytolytic activity
Cell
(2015)
F.S. Hodi et al.
Combined nivolumab and ipilimumab versus ipilimumab alone in patients with advanced melanoma: 2-year overall survival outcomes in a multicentre, randomised, controlled, phase 2 trial
Lancet Oncol
(2016)
N.R. Maimela et al.
Fates of CD8+ T cells in Tumor Microenvironment
Comput Struct Biotechnol J
(2019)
P. Sharma et al.
Primary, Adaptive, and Acquired Resistance to Cancer Immunotherapy
Cell
(2017)
M.C. Mihm et al.
Reflections on the Histopathology of Tumor-Infiltrating Lymphocytes in Melanoma and the Host Immune Response
Cancer Immunol Res
(2015)
F.S. Hodi et al.
Improved survival with ipilimumab in patients with metastatic melanoma
N. Engl. J. Med
(2010)
P. Ghatalia et al.
Approved checkpoint inhibitors in bladder cancer: which drug should be used when?
Ther Adv Med Oncol
(2018)
M. Yarchoan et al.
Tumor Mutational Burden and Response Rate to PD-1 Inhibition
N Engl J Med
(2017)
E.M. Van Allen et al.
Genomic correlates of response to CTLA-4 blockade in metastatic melanoma
Science
(2015)

Krueger F. Trim Galore [Internet];...

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FULL-LENGTH ARTICLEClinical StudiesCommon phenotypic dynamics of tumor-infiltrating lymphocytes across different histologies upon checkpoint inhibition: impact on clinical outcome

Abstract

Background

Aim

Methods

Results

Conclusions

Graphical Abstract

Section snippets

Background

Study design

Patients characteristics

Discussion

Funding

Disclosure of Competing Interest

Author Contributions

Korean J Intern Med

Cell

Lancet Oncol

Comput Struct Biotechnol J

Cell

Reflections on the Histopathology of Tumor-Infiltrating Lymphocytes in Melanoma and the Host Immune Response

Cancer Immunol Res

Improved survival with ipilimumab in patients with metastatic melanoma

N. Engl. J. Med

Approved checkpoint inhibitors in bladder cancer: which drug should be used when?

Ther Adv Med Oncol

Tumor Mutational Burden and Response Rate to PD-1 Inhibition

N Engl J Med

Genomic correlates of response to CTLA-4 blockade in metastatic melanoma

Science

Cancer immunology. Mutational landscape determines sensitivity to PD-1 blockade in non-small cell lung cancer

Science

Evolution of Neoantigen Landscape during Immune Checkpoint Blockade in Non-Small Cell Lung Cancer

Cancer Discov

PD-L1 expression in human cancers and its association with clinical outcomes

Onco Targets Ther

Mutations Associated with Acquired Resistance to PD-1 Blockade in Melanoma

N Engl J Med

Predictive correlates of response to the anti-PD-L1 antibody MPDL3280A in cancer patients

Nature

Elements of cancer immunity and the cancer-immune set point

Nature

Tumor and Microenvironment Evolution during Immunotherapy with Nivolumab

Cell

Chromatin states define tumour-specific T cell dysfunction and reprogramming

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Cancer immunoediting: integrating immunity's roles in cancer suppression and promotion

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Cancer immunoediting: from immunosurveillance to tumor escape

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Rescue of exhausted CD8 T cells by PD-1-targeted therapies is CD28-dependent

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Immune-checkpoint inhibitors for combating T-cell dysfunction in cancer

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A transcriptionally and functionally distinct PD-1(+) CD8(+) T cell pool with predictive potential in non-small-cell lung cancer treated with PD-1 blockade

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PD-1 identifies the patient-specific CD8(+) tumor-reactive repertoire infiltrating human tumors

J Clin Invest

Bystander CD8(+) T cells are abundant and phenotypically distinct in human tumour infiltrates

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FULL-LENGTH ARTICLE
Clinical Studies
Common phenotypic dynamics of tumor-infiltrating lymphocytes across different histologies upon checkpoint inhibition: impact on clinical outcome