Clinicopathological implications of TIM3+ tumor-infiltrating lymphocytes and the miR-455-5p/Galectin-9 axis in skull base chordoma patients
Chordoma is difficult to eradicate due to high local recurrence rates. The immune microenvironment is closely associated with tumor prognosis; however, its role in skull base chordoma is unknown. The expression of Galectin-9 (Gal9) and tumor-infiltrating lymphocyte (TIL) markers was assessed by immunohistochemistry. Kaplan–Meier and multivariate Cox analyses were used to assessing local recurrence-free survival (LRFS) and overall survival (OS) of patients. MiR-455-5p was identified as a regulator of Gal9 expression. Immunopositivity for Gal9 was associated with tumor invasion (p = 0.019), Karnofsky performance status (KPS) score (p = 0.017), and total TIL count (p < 0.001); downregulation of miR-455-5p was correlated with tumor invasion (p = 0.017) and poor prognosis; and the T-cell immunoglobulin and mucin-domain 3 (TIM3)+ TIL count was associated with chordoma invasion (p = 0.010) and KPS score (p = 0.037). Furthermore, multivariate analysis indicated that only TIM3+ TIL density was an independent prognostic factor for LRFS (p = 0.010) and OS (p = 0.016). These results can be used to predict clinical outcome and provide a basis for immune therapy in skull base chordoma patients.
KeywordsSkull base chordoma TIM3 Galectin–9 CD8 FOXp3 miR-455-5p
American Type Culture Collection
C-Casitas B lineage lymphoma
Cluster of differentiation 8
Cytolytic T lymphocyte-associated Ag-4
Hematoxylin and eosin
Karnofsky performance status
Local recurrence-free survival
Quantitative real-time polymerase chain reaction
Sex-determining region Y (SRY)-box 9
T-cell immunoglobulin and mucin-domain 3
ZJ and YZ conceived and designed the study; JZ and YJ performed the experiments and collected the data; HZ, JZ, LC, PL, LL, JZ, and YJ produced the figures and tables; all authors performed the analysis and analyzed the data. JZ, YJ, YZ, and ZJ interpreted results and wrote the manuscript. FL, DZ, and HZ modified the manuscript. All authors read and approved the final version of the manuscript.
This work was supported by the National Natural Science Foundation of China (Nos. 81101917, 81270036, 30901736), the Liaoning Province Natural Science Foundation (No. 20170541022), the Plan to Focus on Research and Development from Science and Technology project of Liaoning Province (No. 2017225029), the Science and Technology Plan Project of Shenyang City (No. 18-014-4-11), and the Fund for Scientific Research of The First Hospital of China Medical University (No. FHCMU-FSR).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interests.
The study was approved by the Research Ethics Committee of the First Hospital of China Medical University and was in accordance with the ethical standards of the institutional committees and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The study approval number is AF-SOP-07-01.
Informed consent was obtained from all individual participants included in the study. With approval from the National Science Foundation of China (81101917), we designed informed consent forms that were signed by eligible patients before recruitment into the study and admission to the hospital. The patients agreed to the use of their specimens and clinical data for research purposes only.
Cell line authentication
The human chordoma cell line UM-Chor1 was obtained as a gift from Professor Yazhuo Zhang, Department of Neurosurgery, Beijing Tiantan Hospital. The origin of UM-Chor1 was human clivus chordoma tissue. The American Type Culture Collection (ATCC) performed authentication of cell line UM-Chor1 via cytochrome C oxidase I assay and short tandem repeat analysis. We obtained a cell line authentication certificate from the ATCC.
- 1.McMaster ML, Goldstein AM, Bromley CM, Ishibe N, Parry DM (2001) Chordoma: incidence and survival patterns in the United States, 1973–1995. Cancer Causes Control 12(1):1–11Google Scholar
- 2.Heffelfinger MJ, Dahlin DC, MacCarty CS, Beabout JW (1973) Chordomas and cartilaginous tumors at the skull base. Cancer 32(2):410–420Google Scholar
- 5.Chibbaro S, Cornelius JF, Froelich S, Tigan L, Kehrli P, Debry C, Romano A, Herman P, George B, Bresson D (2014) Endoscopic endonasal approach in the management of skull base chordomas—clinical experience on a large series, technique, outcome, and pitfalls. Neurosurg Rev 37(2):217–224. https://doi.org/10.1007/s10143-013-0503-9 (discussion 224–215) Google Scholar
- 8.Koutourousiou M, Gardner PA, Tormenti MJ, Henry SL, Stefko ST, Kassam AB, Fernandez-Miranda JC, Snyderman CH (2012) Endoscopic endonasal approach for resection of cranial base chordomas: outcomes and learning curve. Neurosurgery 71(3):614–624. https://doi.org/10.1227/neu.0b013e31825ea3e0 (discussion 624–615) Google Scholar
- 10.Dougan M, Dranoff G (2009) Immune therapy for cancer. Annu Rev Immunol 27:83–117. https://doi.org/10.1146/annurev.immunol.021908.132544 Google Scholar
- 12.Garber ST, Hashimoto Y, Weathers SP, Xiu J, Gatalica Z, Verhaak RG, Zhou S, Fuller GN, Khasraw M, de Groot J, Reddy SK, Spetzler D, Heimberger AB (2016) Immune checkpoint blockade as a potential therapeutic target: surveying CNS malignancies. Neurooncology 18(10):1357–1366. https://doi.org/10.1093/neuonc/now132 Google Scholar
- 16.Hiraoka K, Miyamoto M, Cho Y, Suzuoki M, Oshikiri T, Nakakubo Y, Itoh T, Ohbuchi T, Kondo S, Katoh H (2006) Concurrent infiltration by CD8+ T cells and CD4+ T cells is a favourable prognostic factor in non-small-cell lung carcinoma. Br J Cancer 94(2):275–280. https://doi.org/10.1038/sj.bjc.6602934 Google Scholar
- 18.Seo AN, Lee HJ, Kim EJ, Kim HJ, Jang MH, Lee HE, Kim YJ, Kim JH, Park SY (2013) Tumour-infiltrating CD8+ lymphocytes as an independent predictive factor for pathological complete response to primary systemic therapy in breast cancer. Br J Cancer 109(10):2705–2713. https://doi.org/10.1038/bjc.2013.634 Google Scholar
- 19.Li G, Wang Z, Zhang C, Liu X, Cai J, Wang Z, Hu H, Wu F, Bao Z, Liu Y, Zhao L, Liang T, Yang F, Huang R, Zhang W, Jiang T (2017) Molecular and clinical characterization of TIM-3 in glioma through 1,024 samples. Oncoimmunology 6(8):e1328339. https://doi.org/10.1080/2162402X.2017.1328339 Google Scholar
- 22.Monney L, Sabatos CA, Gaglia JL, Ryu A, Waldner H, Chernova T, Manning S, Greenfield EA, Coyle AJ, Sobel RA, Freeman GJ, Kuchroo VK (2002) Th1-specific cell surface protein Tim-3 regulates macrophage activation and severity of an autoimmune disease. Nature 415(6871):536–541. https://doi.org/10.1038/415536a Google Scholar
- 29.Komita H, Koido S, Hayashi K, Kan S, Ito M, Kamata Y, Suzuki M, Homma S (2015) Expression of immune checkpoint molecules of T cell immunoglobulin and mucin protein 3/galectin-9 for NK cell suppression in human gastrointestinal stromal tumors. Oncol Rep 34(4):2099–2105. https://doi.org/10.3892/or.2015.4149 Google Scholar
- 39.Zou MX, Peng AB, Lv GH, Wang XB, Li J, She XL, Jiang Y (2016) Expression of programmed death-1 ligand (PD-L1) in tumor-infiltrating lymphocytes is associated with favorable spinal chordoma prognosis. Am J Transl Res 8(7):3274–3287Google Scholar
- 40.Bellmunt J, Mullane SA, Werner L, Fay AP, Callea M, Leow JJ, Taplin ME, Choueiri TK, Hodi FS, Freeman GJ, Signoretti S (2015) Association of PD-L1 expression on tumor-infiltrating mononuclear cells and overall survival in patients with urothelial carcinoma. Ann Oncol 26(4):812–817. https://doi.org/10.1093/annonc/mdv009 Google Scholar
- 41.Hutterer M, Knyazev P, Abate A, Reschke M, Maier H, Stefanova N, Knyazeva T, Barbieri V, Reindl M, Muigg A, Kostron H, Stockhammer G, Ullrich A (2008) Axl and growth arrest specific gene 6 are frequently overexpressed in human gliomas and predict poor prognosis in patients with glioblastoma multiforme. Clin Cancer Res 14(1):130–138. https://doi.org/10.1158/1078-0432.ccr-07-0862 Google Scholar
- 43.Sato E, Olson SH, Ahn J, Bundy B, Nishikawa H, Qian F, Jungbluth AA, Frosina D, Gnjatic S, Ambrosone C, Kepner J, Odunsi T, Ritter G, Lele S, Chen YT, Ohtani H, Old LJ, Odunsi K (2005) Intraepithelial CD8+ tumor-infiltrating lymphocytes and a high CD8+/regulatory T cell ratio are associated with favorable prognosis in ovarian cancer. Proc Natl Acad Sci USA 102(51):18538–18543. https://doi.org/10.1073/pnas.0509182102 Google Scholar
- 50.Imaizumi T, Kumagai M, Sasaki N, Kurotaki H, Mori F, Seki M, Nishi N, Fujimoto K, Tanji K, Shibata T, Tamo W, Matsumiya T, Yoshida H, Cui XF, Takanashi S, Hanada K, Okumura K, Yagihashi S, Wakabayashi K, Nakamura T, Hirashima M, Satoh K (2002) Interferon-gamma stimulates the expression of galectin-9 in cultured human endothelial cells. J Leukoc Biol 72(3):486–491Google Scholar
- 51.Yoshida H, Imaizumi T, Kumagai M, Kimura K, Satoh C, Hanada N, Fujimoto K, Nishi N, Tanji K, Matsumiya T, Mori F, Cui XF, Tamo W, Shibata T, Takanashi S, Okumura K, Nakamura T, Wakabayashi K, Hirashima M, Sato Y, Satoh K (2001) Interleukin-1beta stimulates galectin-9 expression in human astrocytes. Neuroreport 12(17):3755–3758Google Scholar
- 52.Sabatos CA, Chakravarti S, Cha E, Schubart A, Sanchez-Fueyo A, Zheng XX, Coyle AJ, Strom TB, Freeman GJ, Kuchroo VK (2003) Interaction of Tim-3 and Tim-3 ligand regulates T helper type 1 responses and induction of peripheral tolerance. Nat Immunol 4(11):1102–1110. https://doi.org/10.1038/ni988 Google Scholar
- 53.Sanchez-Fueyo A, Tian J, Picarella D, Domenig C, Zheng XX, Sabatos CA, Manlongat N, Bender O, Kamradt T, Kuchroo VK, Gutierrez-Ramos JC, Coyle AJ, Strom TB (2003) Tim-3 inhibits T helper type 1-mediated auto- and alloimmune responses and promotes immunological tolerance. Nat Immunol 4(11):1093–1101. https://doi.org/10.1038/ni987 Google Scholar
- 54.Cheng G, Li M, Wu J, Ji M, Fang C, Shi H, Zhu D, Chen L, Zhao J, Shi L, Xu B, Zheng X, Wu C, Jiang J (2015) Expression of Tim-3 in gastric cancer tissue and its relationship with prognosis. Int J Clin Exp Pathol 8(8):9452–9457Google Scholar
- 55.Li H, Wu K, Tao K, Chen L, Zheng Q, Lu X, Liu J, Shi L, Liu C, Wang G, Zou W (2012) Tim-3/galectin-9 signaling pathway mediates T-cell dysfunction and predicts poor prognosis in patients with hepatitis B virus-associated hepatocellular carcinoma. Hepatology 56(4):1342–1351. https://doi.org/10.1002/hep.25777 Google Scholar
- 65.Fritzsching B, Fellenberg J, Moskovszky L, Sapi Z, Krenacs T, Machado I, Poeschl J, Lehner B, Szendroi M, Bosch AL, Bernd L, Csoka M, Mechtersheimer G, Ewerbeck V, Kinscherf R, Kunz P (2015) CD8(+)/FOXP3(+)-ratio in osteosarcoma microenvironment separates survivors from non-survivors: a multicenter validated retrospective study. Oncoimmunology 4(3):e990800. https://doi.org/10.4161/2162402X.2014.990800 Google Scholar