Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2024 Jul 5;29(7):619-628.
doi: 10.1093/oncolo/oyae054.

Automated Quantitative CD8+ Tumor-Infiltrating Lymphocytes and Tumor Mutation Burden as Independent Biomarkers in Melanoma Patients Receiving Front-Line Anti-PD-1 Immunotherapy

Dylan Fortman  1 , Arivarasan Karunamurthy  2   3 , Douglas Hartman  3 , Hong Wang  4 , Lindsey Seigh  3 , Ibrahim Abukhiran  3 , Yana G Najjar  5 , Liron Pantanowitz  6 , Hassane M Zarour  5 , John M Kirkwood  5 , Diwakar Davar  5
Affiliations

Automated Quantitative CD8+ Tumor-Infiltrating Lymphocytes and Tumor Mutation Burden as Independent Biomarkers in Melanoma Patients Receiving Front-Line Anti-PD-1 Immunotherapy

Dylan Fortman et al. Oncologist. .

Abstract

Background: CD8+ tumor-infiltrating lymphocyte (TIL) predicts response to anti-PD-(L)1 therapy. However, there remains no standardized method to assess CD8+ TIL in melanoma, and developing a specific, cost-effective, reproducible, and clinically actionable biomarker to anti-PD-(L)1 remains elusive. We report on the development of automatic CD8+ TIL density quantification via whole slide image (WSI) analysis in advanced melanoma patients treated with front-line anti-PD-1 blockade, and correlation immunotherapy response.

Methods: Seventy-eight patients treated with PD-1 inhibitors in the front-line setting between January 2015 and May 2023 at the University of Pittsburgh Cancer Institute were included. CD8+ TIL density was quantified using an image analysis algorithm on digitized WSI. Targeted next-generation sequencing (NGS) was performed to determine tumor mutation burden (TMB) in a subset of 62 patients. ROC curves were used to determine biomarker cutoffs and response to therapy. Correlation between CD8+ TIL density and TMB cutoffs and response to therapy was studied.

Results: Higher CD8+ TIL density was significantly associated with improved response to front-line anti-PD-1 across all time points measured. CD8+ TIL density ≥222.9 cells/mm2 reliably segregated responders and non-responders to front-line anti-PD-1 therapy regardless of when response was measured. In a multivariate analysis, patients with CD8+ TIL density exceeding cutoff had significantly improved PFS with a trend toward improved OS. Similarly, increasing TMB was associated with improved response to anti-PD-1, and a cutoff of 14.70 Mut/Mb was associated with improved odds of response. The correlation between TMB and CD8+ TIL density was low, suggesting that each represented independent predictive biomarkers of response.

Conclusions: An automatic digital analysis algorithm provides a standardized method to quantify CD8+ TIL density, which predicts response to front-line anti-PD-1 therapy. CD8+ TIL density and TMB are independent predictors of response to anti-PD-1 blockade.

Keywords: PD-1; PD-L1; TIL; TMB/Mb; biomarker; immunotherapy; melanoma; metastatic; tumor mutation burden.

PubMed Disclaimer

Conflict of interest statement

Hassane M. Zarour: grants/research support (institutional): NIH/NCI, Bristol-Myers Squibb and GlaxoSmithKline. Consultant: Checkmate Pharmaceuticals, GlaxoSmithKline, Bayer, and Vedanta. Intellectual Property: US Patent 63/124,231, “Compositions and Methods for Treating Cancer,” Dec 11, 2020. US Patent 63/208,719, “Compositions and Methods For Responsiveness to Immune Checkpoint Inhibitors (ICI), Increasing Effectiveness of ICI and Treating Cancer,” June 9, 2021. John M. Kirkwood: Grants/Research Support (institutional): Bristol-Myers Squibb, Amgen Inc. Consultant: Bristol-Myers Squibb, Checkmate Pharmaceuticals, Novartis, Amgen Inc., Checkmate, Castle Biosciences, Inc., Immunocore LLC, Iovance, Novartis. Diwakar Davar: Grants/Research Support (institutional): Arcus, CellSight Technologies, Immunocore, Merck, Regeneron Pharmaceuticals Inc., Tesaro/GSK. Consultant: ACM Bio, Ascendis Pharma, Clinical Care Options (CCO), Gerson Lehrman Group (GLG), Merck, Medical Learning Group (MLG), Xilio Therapeutics. CE Speakers’ Bureau: Castle Biosciences. Stockholder: None. Intellectual Property: US Patent 63/124,231, “Compositions and Methods for Treating Cancer,” Dec 11, 2020. US Patent 63/208,719, “Compositions and Methods For Responsiveness to Immune Checkpoint Inhibitors (ICI), Increasing Effectiveness of ICI and Treating Cancer,” June 9, 2021. The other authors indicated no financial relationships.

Figures

Figure 1.
Figure 1.
Correlation between WSI and tumor CD8+ TIL infiltrate in PD-1 treated melanoma patients, and derivation of 222.9 cells/mm2 Cutoff.
Figure 2.
Figure 2.
Relationship between elevated CD8+ TIL and PFS and OS in PD-1 treated melanoma.
Figure 3.
Figure 3.
Derivation of 14.7 Mut/Mb TMB cutoff in anti-PD-1 treated melanoma patients.
Figure 4.
Figure 4.
Relationship between elevated TMB and PFS and OS in PD-1 treated melanoma.
See this image and copyright information in PMC

References

    1. Dong H, Zhu G, Tamada K, Chen L.. B7-H1, a third member of the B7 family, co-stimulates T-cell proliferation and interleukin-10 secretion. Nat Med. 1999;5(12):1365-1369. 10.1038/70932 - DOI - PubMed
    1. Freeman GJ, Long AJ, Iwai Y, et al.. Engagement of the PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med. 2000;192(7):1027-1034. 10.1084/jem.192.7.1027 - DOI - PMC - PubMed
    1. Latchman Y, Wood CR, Chernova T, et al.. PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol. 2001;2(3):261-268. 10.1038/85330 - DOI - PubMed
    1. Tseng SY, Otsuji M, Gorski K, et al.. B7-DC, a new dendritic cell molecule with potent costimulatory properties for T cells. J Exp Med. 2001;193(7):839-846. 10.1084/jem.193.7.839 - DOI - PMC - PubMed
    1. Fourcade J, Kudela P, Sun Z, et al.. PD-1 is a regulator of NY-ESO-1-specific CD8+ T cell expansion in melanoma patients. J Immunol. 2009;182(9):5240-5249. 10.4049/jimmunol.0803245 - DOI - PMC - PubMed

Substances