iNOS Expression by Tumor-Infiltrating Lymphocytes, PD-L1 and Prognosis in Non-Small-Cell Lung Cancer
Abstract
:Simple Summary
Abstract
1. Introduction
2. Results
2.1. Expression of iNOS in Normal Lung
2.2. Expression of iNOS in Cancer
2.3. Association of iNOS Expression with Histopathological Parameters
2.4. Correlation of iNOS Expression with Hypoxic Markers
2.5. Correlation of iNOS Expression with Immunological Parameters
2.6. Survival Analysis
3. Discussion
4. Materials and Methods
4.1. Patient and Disease Characteristics
4.2. Ethical Considerations
4.3. iNOS Immunohistochemistry
4.4. Assessment of Other Immunohistochemical Markers
4.5. Scoring of TIL and iNOS+TIL Density
4.6. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Conflicts of Interest
References
- Michel, T.; Feron, O. Nitric oxide synthases: Which, where, how, and why? J. Clin. Investig. 1997, 100, 2146–2152. [Google Scholar] [CrossRef] [Green Version]
- Alderton, W.K.; Cooper, C.E.; Knowles, R.G. Nitric oxide synthases: Structure, function and inhibition. Biochem. J. 2001, 357, 593–615. [Google Scholar] [CrossRef]
- Aranda, E.; López-Pedrera, C.; De La Haba-Rodriguez, J.R.; Rodriguez-Ariza, A. Nitric oxide and cancer: The emerging role of S-nitrosylation. Curr. Mol. Med. 2012, 12, 50–67. [Google Scholar] [CrossRef] [PubMed]
- Garrido, P.; Shalaby, A.; Walsh, E.M.; Keane, N.; Webber, M.; Keane, M.M.; Sullivan, F.J.; Kerin, M.J.; Callagy, G.; Ryan, A.E.; et al. Impact of inducible nitric oxide synthase (iNOS) expression on triple negative breast cancer outcome and activation of EGFR and ERK signaling pathways. Oncotarget 2017, 8, 80568–80588. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ambs, S.; Hussain, S.P.; Harris, C.C. Interactive effects of nitric oxide and the p53 tumor suppressor gene in carcinogenesis and tumor progression. FASEB J. 1997, 11, 443–448. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Ridnour, L.A.; Thomas, D.D.; Switzer, C.; Flores-Santana, W.; Isenberg, J.S.; Ambs, S.; Roberts, D.D.; Wink, D.A. Molecular mechanisms for discrete nitric oxide levels in cancer. Nitric Oxide 2008, 19, 73–76. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Xiao, L.; Eneroth, P.H.; Qureshi, G.A. Nitric oxide synthase pathway may mediate human natural killer cell cytotoxicity. Scand. J. Immunol. 1995, 42, 505–511. [Google Scholar] [CrossRef] [PubMed]
- Palmieri, E.M.; Gonzalez-Cotto, M.; Baseler, W.A.; Davies, L.C.; Ghesquière, B.; Maio, N.; Rice, C.M.; Rouault, T.A.; Cassel, T.; Higashi, R.M.; et al. Nitric oxide orchestrates metabolic rewiring in M1 macrophages by targeting aconitase 2 and pyruvate dehydrogenase. Nat. Commun. 2020, 11, 698. [Google Scholar] [CrossRef] [PubMed]
- Xue, Q.; Yan, Y.; Zhang, R.; Xiong, H. Regulation of iNOS on Immune Cells and Its Role in Diseases. Int. J. Mol. Sci. 2018, 19, 3805. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Lisanti, M.P.; Tsirigos, A.; Pavlides, S.; Reeves, K.J.; Peiris-Pagès, M.; Chadwick, A.L.; Sanchez-Alvarez, R.; Lamb, R.; Howell, A.; Martinez-Outschoorn, U.E.; et al. JNK1 stress signaling is hyper-activated in high breast density and the tumor stroma: Connecting fibrosis, inflammation, and stemness for cancer prevention. Cell Cycle 2014, 13, 580–599. [Google Scholar] [CrossRef] [Green Version]
- Müerköster, S.; Wegehenkel, K.; Arlt, A.; Witt, M.; Sipos, B.; Kruse, M.-L.; Sebens, T.; Klöppel, G.; Kalthoff, H.; Fölsch, U.R.; et al. Tumor stroma interactions induce chemoresistance in pancreatic ductal carcinoma cells involving increased secretion and paracrine effects of nitric oxide and interleukin-1beta. Cancer Res. 2004, 64, 1331–1337. [Google Scholar] [CrossRef] [Green Version]
- Liao, W.; Ye, T.; Liu, H. Prognostic value of inducible nitric oxide synthase (iNOS) in human cancer: A systematic review and meta-analysis. BioMed Res. Int. 2019, 2019, 6304851. [Google Scholar] [CrossRef] [Green Version]
- Honkanen, T.J.; Tikkanen, A.; Karihtala, P.; Mäkinen, M.; Väyrynen, J.P.; Koivunen, J.P. Prognostic and predictive role of tumour-associated macrophages in HER2 positive breast cancer. Sci. Rep. 2019, 9, 10961. [Google Scholar] [CrossRef] [Green Version]
- Jackute, J.; Zemaitis, M.; Pranys, D.; Sitkauskiene, B.; Miliauskas, S.; Vaitkiene, S.; Sakalauskas, R. Distribution of M1 and M2 macrophages in tumor islets and stroma in relation to prognosis of non-small cell lung cancer. BMC Immunol. 2018, 19, 3. [Google Scholar] [CrossRef] [Green Version]
- Jayasurya, A.; Dheen, S.T.; Yap, W.M.; Tan, N.G.; Ng, Y.K.; Bay, B.H. Inducible nitric oxide synthase and bcl-2 expression in nasopharyngeal cancer: Correlation with outcome of patients after radiotherapy. Int. J. Radiat. Oncol. Biol. Phys. 2003, 56, 837–845. [Google Scholar] [CrossRef]
- Anttila, M.A.; Voutilainen, K.; Merivalo, S.; Saarikoski, S.; Kosma, V.M. Prognostic significance of iNOS in epithelial ovarian cancer. Gynecol. Oncol. 2007, 105, 97–103. [Google Scholar] [CrossRef] [PubMed]
- Puhakka, A.; Kinnula, V.; Näpänkangas, U.; Säily, M.; Koistinen, P.; Pääkkö, P.; Soini, Y. High expression of nitric oxide synthases is a favorable prognostic sign in non- small cell lung carcinoma. APMIS 2003, 111, 1137–1146. [Google Scholar] [CrossRef]
- Rubio, L.; Vera-Sempere, F.J.; Lopez-Guerrero, J.A.; Padilla, J.; Moreno-Baylach, M.J. A risk model for non-small cell lung cancer using clinicopathological variables, angiogenesis and oncoprotein expression. Anticancer Res. 2005, 25, 497–504. [Google Scholar] [PubMed]
- Giatromanolaki, A.; Koukourakis, I.M.; Balaska, K.; Mitrakas, A.G.; Harris, A.L.; Koukourakis, M.I. Programmed death-1 receptor (PD-1) and PD-ligand-1 (PD-L1) expression in non-small cell lung cancer and the immune-suppressive effect of anaerobic glycolysis. Med. Oncol. 2019, 36, 76. [Google Scholar] [CrossRef] [PubMed]
- Jianjun, Y.; Zhang, R.; Lu, G.; Shen, Y.; Peng, L.; Zhu, C.; Cui, M.; Wang, W.; Arnaboldi, P.; Tang, M.; et al. T cell–derived inducible nitric oxide synthase switches off Th17 cell differentiation. J. Exp. Med. 2013, 210, 1447–1462. [Google Scholar]
- Scheller, L.F.; Green, S.J.; Azad, A.F. Inhibition of nitric oxide interrupts the accumulation of CD8+ T cells surrounding Plasmodium berghei-infected hepatocytes. Infect. Immun. 1997, 65, 3882–3888. [Google Scholar] [CrossRef] [Green Version]
- Bogen, B.; Fauskanger, M.; Haabeth, O.A.; Tveita, A. CD4(+) T cells indirectly kill tumor cells via induction of cytotoxic macrophages in mouse models. Cancer Immunol. Immunother. 2019, 68, 1865–1873. [Google Scholar] [CrossRef]
- Jayaraman, P.; Alfarano, M.G.; Svider, P.F.; Parikh, F.; Lu, G.; Kidwai, S.; Xiong, H.; Sikora, A.G. iNOS expression in CD4+ T cells limits Treg induction by repressing TGFβ1: Combined iNOS inhibition and Treg depletion unmask endogenous antitumor immunity. Clin. Cancer Res. 2014, 20, 6439–6451. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Giatromanolaki, A.; Sivridis, E.; Arelaki, S.; Koukourakis, M.I. Expression of enzymes related to glucose metabolism in non-small cell lung cancer and prognosis. Exp. Lung Res. 2017, 43, 167–174. [Google Scholar] [CrossRef]
- Giatromanolaki, A.; Koukourakis, M.I.; Sivridis, E.; Pastorek, J.; Wykoff, C.C.; Gatter, K.C.; Harris, A.L. Expression of hypoxia-inducible carbonic anhydrase-9 relates to angiogenic pathways and independently to poor outcome in non-small cell lung cancer. Cancer Res. 2001, 61, 7992–7998. [Google Scholar]
Extent of Cancer Cell expression | ||
Pattern | % cancer cell reactivity | No pts (%) |
Negative | 0 | 55 (56.1%) |
Low | 1–9 | 24 (24.5%) |
Medium/High | 10–100 | 19 (19.4%) |
Extent of expression by Cancer-Associated Fibroblasts | ||
Pattern | % stroma reactivity | No pts (%) |
Negative | 0 | 83 (84.7) |
Low | 1–9 | 7 (7.1) |
Medium/High | 10–100 | 8 (8.2) |
Percentage of iNOS+TILs | ||
Pattern | % TILs | No pts (%) |
Negative | 0 | 42 (42.8%) |
Low | 1–9 | 33 (33.7%) |
Medium/High | 10–100 | 23 (23.5%) |
iNOS+TIL score | ||
Pattern | % TILs | No pts (%) |
0 | 0 | 42 (42.9%) |
1 | 0.04–0.1 | 9 (9.2%) |
2 | 0.15–0.3 | 23 (23.5%) |
3 | 0.4–3.6 | 24 (24.5%) |
Histology | ||||
Cell Type | S | A | L | p-value |
Cancer Cells | ||||
Negative (55) | 26 | 17 | 12 | 0.02 (Sq vs. other) |
Low (24) | 17 | 4 | 3 | |
Med/High (19) | 15 | 1 | 3 | |
Stroma CAFs | ||||
Neg (83) | 45 | 21 | 17 | 0.01 (Sq vs. other) |
Low (7) | 6 | 0 | 1 | |
Medium/High (8) | 7 | 1 | 0 | |
iNOS+TIL score | ||||
0/1 (51) | 25 | 14 | 12 | 0.03 (Sq vs. other) |
2/3 (47) | 33 | 8 | 6 | |
Stage | ||||
Cell Type | 1 | 2 | 3 | p-value |
Cancer Cells | ||||
Negative (55) | 27 | 10 | 18 | 0.61 |
Low (24) | 12 | 7 | 5 | |
Med/High (19) | 7 | 5 | 7 | |
Stroma CAFs | ||||
Neg (83) | 40 | 18 | 25 | 0.83 |
Low (7) | 3 | 2 | 2 | |
Medium/High (8) | 3 | 2 | 2 | |
iNOS+TIL score | ||||
0/1 (51) | 24 | 11 | 16 | 0.97 |
2/3 (47) | 22 | 11 | 14 |
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Giatromanolaki, A.; Tsolou, A.; Daridou, E.; Kouroupi, M.; Chlichlia, K.; Koukourakis, M.I. iNOS Expression by Tumor-Infiltrating Lymphocytes, PD-L1 and Prognosis in Non-Small-Cell Lung Cancer. Cancers 2020, 12, 3276. https://doi.org/10.3390/cancers12113276
Giatromanolaki A, Tsolou A, Daridou E, Kouroupi M, Chlichlia K, Koukourakis MI. iNOS Expression by Tumor-Infiltrating Lymphocytes, PD-L1 and Prognosis in Non-Small-Cell Lung Cancer. Cancers. 2020; 12(11):3276. https://doi.org/10.3390/cancers12113276
Chicago/Turabian StyleGiatromanolaki, Alexandra, Avgi Tsolou, Eleftheria Daridou, Maria Kouroupi, Katerina Chlichlia, and Michael I. Koukourakis. 2020. "iNOS Expression by Tumor-Infiltrating Lymphocytes, PD-L1 and Prognosis in Non-Small-Cell Lung Cancer" Cancers 12, no. 11: 3276. https://doi.org/10.3390/cancers12113276