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
. 2017 Dec 19;9(3):3805-3814.
doi: 10.18632/oncotarget.23454. eCollection 2018 Jan 9.

Similarity and difference in tumor-infiltrating lymphocytes in original tumor tissues and those of in vitro expanded populations in head and neck cancer

Lili Ren  1   2 , Tatsuo Matsuda  1 , Boya Deng  1 , Kazuma Kiyotani  3 , Taigo Kato  1 , Jae-Hyun Park  1 , Tanguy Y Seiwert  1 , Everett E Vokes  1 , Nishant Agrawal  4 , Yusuke Nakamura  1   4
Affiliations

Similarity and difference in tumor-infiltrating lymphocytes in original tumor tissues and those of in vitro expanded populations in head and neck cancer

Lili Ren et al. Oncotarget. .

Abstract

Though adoptive tumor-infiltrating lymphocyte (TIL) therapy has been explored in clinical trials for many years, there is little information for the clonotype composition between TILs in original tumor tissues and TILs that were in vitro expanded and infused to cancer patients. To investigate the similarity/difference in TILs in original tumor tissues and those of in vitro expanded populations in squamous cell carcinoma of head and neck (SCCHN) as well as their correlation with somatic mutations in cancer cells, we performed whole exome analysis, expression profile analysis of immune-related genes, and T cell receptor (TCR) analysis of original TILs and in vitro expanded TILs in 8 surgically-resected HPV-negative fresh tumors with SCCHN. We found an unusually high number of non-synonymous somatic mutations (4290, 1779 and 901 mutations) in three SCCHN tumors, in which we identified mutations in mismatch repair genes, MSH2 or MSH4, or a DNA polymerase gene, POLE. Interestingly, dominant TCR clonotypes of expanded CD8+ TILs derived from these three tumors revealed high similarity to those in original tumors while for remaining tumors with the lower mutational load, we found that T cell clonotypes between TILs in original tumor tissues and those expanded in vitro were almost entirely different. Our findings might provide clinically useful information for identification of tumor-antigen-specific T cell clones that may lead to further improvement of adoptive TIL therapy for SCCHN patients.

Keywords: T cell receptor; mismatch repair; non-synonymous mutation; squamous cell carcinoma of head and neck cancer; tumor-infiltrating lymphocytes.

PubMed Disclaimer

Conflict of interest statement

CONFLICTS OF INTEREST Y. N. is a stock holder and a scientific advisor of OncoTherapy Science, Inc. J-H. Park is a scientific advisor of OncoTherapy Science, Inc. No potential conflicts of interest were disclosed by the other authors.

Figures

Figure 1
Figure 1. Expression profiles of immune-related genes in tumor microenvironment
(A) Heatmap of mRNA expression levels of 11 immune-related genes, TRB, CD4, CD8, FOXP3, PD-1, OX40, GZMA, PRF1, PD-L1, IL10, and TIM3 normalized by GAPDH expression levels in 8 SCCHN tumors. Correlation analysis of expression levels: GZMA to FOXP3 (B) and OX40 to GZMA (C).
Figure 2
Figure 2
(A) Distribution of both TCR-α and TCR-β CDR3 unique clonotypes, the numbers of mutations in individual tumors as well as TCR-β DIs in original TILs in SCCHN tumors. (B) Distribution of both TCR-α and TCR-β CDR3 unique clonotypes and TCR-β DIs in expanded CD4+/CD8+ TILs. Pie charts depict clonotypes with 0.1% frequency or higher in each patient. Common colors among different pie charts do not represent identical clonotypes. The light orange portion of each pie chart contains clonotypes with less than 0.1% frequency.
Figure 3
Figure 3. Comparison of TCR-β repertoire between TILs in original tumors and those expanded in vitro
(A) Comparison of TCR-β clonotypes of TILs before and after expansion in 3 tumors with very high mutational load. (B) Comparison of TCR-β clonotypes of TILs before and after expansion in remaining 4 tumors. The blue bars represent the frequencies (ordered by the frequencies from highest to lowest) of the common CDR3 clonotypes (the frequency of 0.1% or higher in all mapped TCR reads) in TILs in the original tumors (a mixture of CD4+ and CD8+ cells). The orange bars represent the frequencies of CDR3 clonotypes in expanded CD8+ populations and the red bars represent those in the expanded CD4+ populations. (C) Heatmap showing changes between original top 20 clones before and after in vitro expansion. For each patient, TCR-β repertoire data of original TILs (represented by original), expanded CD8+ TILs (represented by EX-CD8), expanded CD4+ TILs (represented by EX-CD4) are listed. The frequencies of individual CDR3 clonotypes are indicated by graduation of colors and different colors on the bottom of the heatmap indicate individual patients.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

References

    1. Global Burden of Disease Cancer Collaboration. Fitzmaurice C, Allen C, Barber RM, Barregard L, Bhutta ZA, Brenner H, Dicker DJ, Chimed-Orchir O, Dandona R, Dandona L, Fleming T, Forouzanfar MH, et al. Global, regional, and national cancer incidence, mortality, years of life lost, years lived with disability, and disability-adjusted life-years for 32 cancer groups, 1990 to 2015: a systematic analysis for the global burden of disease study. JAMA Oncol. 2017;3:524–48. https://doi.org/10.1001/jamaoncol.2016.5688. - DOI - PMC - PubMed
    1. Chaturvedi AK, Anderson WF, Lortet-Tieulent J, Curado MP, Ferlay J, Franceschi S, Rosenberg PS, Bray F, Gillison ML. Worldwide trends in incidence rates for oral cavity and oropharyngeal cancers. J Clin Oncol. 2013;31:4550–9. https://doi.org/10.1200/JCO.2013.50.3870. - DOI - PMC - PubMed
    1. Kitano A, Ono M, Yoshida M, Noguchi E, Shimomura A, Shimoi T, Kodaira M, Yunokawa M, Yonemori K, Shimizu C, Kinoshita T, Fujiwara Y, Tsuda H, et al. Tumour-infiltrating lymphocytes are correlated with higher expression levels of PD-1 and PD-L1 in early breast cancer. ESMO Open. 2017;2:e000150. https://doi.org/10.1136/esmoopen-2016-000150. - DOI - PMC - PubMed
    1. Lechner A, Schlosser H, Rothschild SI, Thelen M, Reuter S, Zentis P, Shimabukuro-Vornhagen A, Theurich S, Wennhold K, Garcia-Marquez M, Tharun L, Quaas A, Schauss A, et al. Characterization of tumor-associated T-lymphocyte subsets and immune checkpoint molecules in head and neck squamous cell carcinoma. Oncotarget. 2017;8:44418–33. https://doi.org/10.18632/oncotarget.17901. - DOI - PMC - PubMed
    1. Tomioka N, Azuma M, Ikarashi M, Yamamoto M, Sato M, Watanabe KI, Yamashiro K, Takahashi M. The therapeutic candidate for immune checkpoint inhibitors elucidated by the status of tumor-infiltrating lymphocytes (TILs) and programmed death ligand 1 (PD-L1) expression in triple negative breast cancer (TNBC) Breast Cancer. 2017 https://doi.org/10.1007/s12282-017-0781-0. - DOI - PubMed