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Targeting platelets for improved outcome in KRAS-driven lung adenocarcinoma

Abstract

Elevated platelet count is associated with poor survival in certain solid cancers, including lung cancer. In addition, experimental transplantation of cancer cell lines has uncovered a role for platelets in blood-borne metastasis. These studies, however, do not account for heterogeneity between lung cancer subtypes. Subsequently, the role of platelets in the major subtypes of non-small cell lung cancer (adenocarcinoma (ADC) and squamous cell carcinoma (SqCC)) is not fully understood. We utilised an autochthonous KrasLSL-G12D/+;p53flox/flox mouse model of lung ADC together with genetic models of thrombocytopenia to interrogate the role of platelets in lung cancer growth and progression. While thrombocytopenia failed to impact primary tumour growth, in experimental metastatic models however, thrombocytopenic mice displayed significantly extended survival. Utilising a novel thrombocytopenic immunocompromised mouse, the importance of platelets in metastatic dissemination was confirmed with human KRAS-mutant ADC cell lines. Finally, retrospective analysis of a NSCLC patient cohort revealed thrombocytosis was predictive of poor survival in ADC patients with metastatic disease. Interestingly, this association was not apparent in SqCC patients. Overall, these data highlight the possibility of patient stratification using thrombocytosis as a biomarker, and indicates opportunities for potential novel treatment strategies that combine anti-platelet and lung cancer therapies.

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Fig. 1: Platelets influence metastasis but not primary growth of murine KRAS-mutant ADC.
Fig. 2: Platelet molecules released during activation influence ADC metastasis.
Fig. 3: Thrombocytosis is indicative of reduced survival in patients with metastatic ADC.

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Data availability

The datasets generated and/or analysed during the current study are available from the corresponding author on reasonable request. Additional methods can be found in the supplementary information.

Code availability

All code used for analysis of lung tumour burden and HMGA2 positive staining cells is available from GitHub (https://github.com/SR-H/Image-analysis-code.git).

References

  1. Chen Z, Fillmore CM, Hammerman PS, Kim CF, Wong KK. Non-small-cell lung cancers: a heterogeneous set of diseases. Nat Rev Cancer. 2014;14:535–46.

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Spiro SG, Silvestri GA. The treatment of advanced non-small cell lung cancer. Curr Opin Pulm Med. 2005;11:287–91.

    CAS  PubMed  Google Scholar 

  3. Coenen DM, Mastenbroek TG, Cosemans J. Platelet interaction with activated endothelium: mechanistic insights from microfluidics. Blood. 2017;130:2819–28.

    CAS  PubMed  Google Scholar 

  4. Hyslop SR, Josefsson EC. Undercover Agents: targeting tumours with modified platelets. Trends Cancer. 2017;3:235–46.

    CAS  PubMed  Google Scholar 

  5. Gay LJ, Felding-Habermann B. Contribution of platelets to tumour metastasis. Nat Rev Cancer. 2011;11:123–34.

    CAS  PubMed  PubMed Central  Google Scholar 

  6. Labelle M, Begum S, Hynes RO. Direct signaling between platelets and cancer cells induces an epithelial-mesenchymal-like transition and promotes metastasis. Cancer Cell. 2011;20:576–90.

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Kopp HG, Placke T, Salih HR. Platelet-derived transforming growth factor-beta down-regulates NKG2D thereby inhibiting natural killer cell antitumor reactivity. Cancer Res. 2009;69:7775–83.

    CAS  PubMed  Google Scholar 

  8. Schumacher D, Strilic B, Sivaraj KK, Wettschureck N, Offermanns S. Platelet-derived nucleotides promote tumor-cell transendothelial migration and metastasis via P2Y2 receptor. Cancer Cell. 2013;24:130–7.

    CAS  PubMed  Google Scholar 

  9. Lucotti S, Cerutti C, Soyer M, Gil-Bernabe AM, Gomes AL, Allen PD, et al. Aspirin blocks formation of metastatic intravascular niches by inhibiting platelet-derived COX-1/thromboxane A2. J Clin Investig. 2019;130:1845–62.

    Google Scholar 

  10. Rothwell PM, Fowkes FG, Belch JF, Ogawa H, Warlow CP, Meade TW. Effect of daily aspirin on long-term risk of death due to cancer: analysis of individual patient data from randomised trials. Lancet. 2011;377:31–41.

    CAS  PubMed  Google Scholar 

  11. McNeil JJ, Nelson MR, Woods RL, Lockery JE, Wolfe R, Reid CM, et al. Effect of aspirin on all-cause mortality in the healthy elderly. N. Engl J Med. 2018;379:1519–28.

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Wang JJ, Wang YL, Ge XX, Xu MD, Chen K, Wu MY, et al. Prognostic values of platelet-associated indicators in resectable lung cancers. Technol Cancer Res Treat. 2019;18:1533033819837261.

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Pedersen LM, Milman N. Prognostic significance of thrombocytosis in patients with primary lung cancer. Eur Respir J. 1996;9:1826–30.

    CAS  PubMed  Google Scholar 

  14. Tomita M, Shimizu T, Hara M, Ayabe T, Onitsuka T. Prognostic impact of thrombocytosis in resectable non-small cell lung cancer. Interact Cardiovasc Thorac Surg. 2008;7:613–5.

    PubMed  Google Scholar 

  15. Levin J, Conley CL. Thrombocytosis associated with malignant disease. Arch Intern Med. 1964;114:497–500.

    CAS  PubMed  Google Scholar 

  16. Roselli M, Mineo TC, Martini F, Mariotti S, Ambrogi V, Spila A, et al. Soluble selectin levels in patients with lung cancer. Int J Biol Markers. 2002;17:56–62.

    CAS  PubMed  Google Scholar 

  17. Jain R, Tabor DC, Engle JC. Plasma beta-thromboglobulin levels in lung cancer. South Med J. 1983;76:1380–2.

    CAS  PubMed  Google Scholar 

  18. Roselli M, Mineo TC, Basili S, Martini F, Mariotti S, Aloe S, et al. Soluble CD40 ligand plasma levels in lung cancer. Clin Cancer Res. 2004;10:610–4.

    CAS  PubMed  Google Scholar 

  19. Jackson EL, Willis N, Mercer K, Bronson RT, Crowley D, Montoya R, et al. Analysis of lung tumor initiation and progression using conditional expression of oncogenic K-ras. Genes Dev. 2001;15:3243–8.

    CAS  PubMed  PubMed Central  Google Scholar 

  20. Jackson EL, Olive KP, Tuveson DA, Bronson R, Crowley D, Brown M, et al. The differential effects of mutant p53 alleles on advanced murine lung cancer. Cancer Res. 2005;65:10280–8.

    CAS  PubMed  Google Scholar 

  21. DuPage M, Dooley AL, Jacks T. Conditional mouse lung cancer models using adenoviral or lentiviral delivery of Cre recombinase. Nat Protoc. 2009;4:1064–72.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Mason KD, Carpinelli MR, Fletcher JI, Collinge JE, Hilton AA, Ellis S, et al. Programmed anuclear cell death delimits platelet life span. Cell. 2007;128:1173–86.

    CAS  PubMed  Google Scholar 

  23. Winslow MM, Dayton TL, Verhaak RG, Kim-Kiselak C, Snyder EL, Feldser DM, et al. Suppression of lung adenocarcinoma progression by Nkx2-1. Nature. 2011;473:101–4.

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Pucci F, Rickelt S, Newton AP, Garris C, Nunes E, Evavold C, et al. PF4 promotes platelet production and lung cancer growth. Cell Rep. 2016;17:1764–72.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Izaguirre-Avila R, De la Pena-Diaz A, Barinagarrementeria-Aldatz F, Gonzalez-Pacheco H, Ramirez-Gutierrez AE, Ruiz-Sandoval JL, et al. Effect of clopidogrel on platelet aggregation and plasma concentration of fibrinogen in subjects with cerebral or coronary atherosclerotic disease. Clin Appl Thromb Hemost. 2002;8:169–77.

    CAS  PubMed  Google Scholar 

  26. Zhang H, Xia H, Zhang L, Zhang B, Yue D, Wang C. Clinical significance of preoperative neutrophil-lymphocyte vs platelet-lymphocyte ratio in primary operable patients with non-small cell lung cancer. Am J Surg. 2015;210:526–35.

    PubMed  Google Scholar 

  27. Yu LX, Yan L, Yang W, Wu FQ, Ling Y, Chen SZ, et al. Platelets promote tumour metastasis via interaction between TLR4 and tumour cell-released high-mobility group box1 protein. Nat Commun. 2014;5:5256.

    CAS  PubMed  Google Scholar 

  28. Mammadova-Bach E, Zigrino P, Brucker C, Bourdon C, Freund M, De Arcangelis A, et al. Platelet integrin alpha6beta1 controls lung metastasis through direct binding to cancer cell-derived ADAM9. JCI Insight 2016;1:e88245.

    PubMed  PubMed Central  Google Scholar 

  29. Gay LJ, Felding-Habermann B. Platelets alter tumor cell attributes to propel metastasis: programming in transit. Cancer Cell. 2011;20:553–4.

    CAS  PubMed  Google Scholar 

  30. Plantureux L, Mege D, Crescence L, Carminita E, Robert S, Cointe S, et al. The interaction of platelets with colorectal cancer cells inhibits tumor growth but promotes metastasis. Cancer Res. 2019;80:291–303.

  31. Michael JV, Wurtzel JGT, Mao GF, Rao AK, Kolpakov MA, Sabri A, et al. Platelet microparticles infiltrating solid tumors transfer miRNAs that suppress tumor growth. Blood. 2017;130:567–80.

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Takagi S, Sato S, Oh-hara T, Takami M, Koike S, Mishima Y, et al. Platelets promote tumor growth and metastasis via direct interaction between Aggrus/podoplanin and CLEC-2. PLoS ONE. 2013;8:e73609.

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Cho MS, Bottsford-Miller J, Vasquez HG, Stone R, Zand B, Kroll MH, et al. Platelets increase the proliferation of ovarian cancer cells. Blood. 2012;120:4869–72.

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Miyata K, Takemoto A, Okumura S, Nishio M, Fujita N. Podoplanin enhances lung cancer cell growth in vivo by inducing platelet aggregation. Sci Rep. 2017;7:4059.

    PubMed  PubMed Central  Google Scholar 

  35. Zhang X, Ran Y. Prognostic role of elevated platelet count in patients with lung cancer: a systematic review and meta-analysis. Int J Clin Exp Med. 2015;8:5379–87.

    PubMed  PubMed Central  Google Scholar 

  36. Niki M, Yokoi T, Kurata T, Nomura S. New prognostic biomarkers and therapeutic effect of bevacizumab for patients with non-small-cell lung cancer. Lung Cancer (Auckl). 2017;8:91–9.

    CAS  Google Scholar 

  37. Osmani L, Askin F, Gabrielson E, Li QK. Current WHO guidelines and the critical role of immunohistochemical markers in the subclassification of non-small cell lung carcinoma (NSCLC): Moving from targeted therapy to immunotherapy. Semin Cancer Biol. 2018;52:103–9.

    CAS  PubMed  Google Scholar 

  38. Malhotra J, Jabbour SK, Aisner J. Current state of immunotherapy for non-small cell lung cancer. Transl Lung Cancer Res. 2017;6:196–211.

    CAS  PubMed  PubMed Central  Google Scholar 

  39. Rachidi S, Metelli A, Riesenberg B, Wu BX, Nelson MH, Wallace C, et al. Platelets subvert T cell immunity against cancer via GARP-TGFβ axis. Sci Immunol. 2017;2:eaai7911. https://doi.org/10.1126/sciimmunol.aai7911.

  40. Riesenberg BP, Ansa-Addo EA, Gutierrez J, Timmers CD, Liu B, Li Z. Cutting Edge: Targeting Thrombocytes to Rewire Anticancer Immunity in the Tumor Microenvironment and Potentiate Efficacy of PD-1 Blockade. J Immunol. 2019;203:1105–10.

    CAS  PubMed  Google Scholar 

  41. Han R, Hao S, Lu C, Zhang C, Lin C, Li L, et al. Aspirin sensitizes osimertinib-resistant NSCLC cells in vitro and in vivo via Bim-dependent apoptosis induction. Mol Oncol. 2020;14:1152–69.

  42. Hu X, Wu LW, Weng X, Lin NM, Zhang C. Synergistic antitumor activity of aspirin and erlotinib: Inhibition of p38 enhanced aspirin plus erlotinib-induced suppression of metastasis and promoted cancer cell apoptosis. Oncol Lett. 2018;16:2715–24.

    PubMed  PubMed Central  Google Scholar 

  43. Alexander WS, Roberts AW, Nicola NA, Li R, Metcalf D. Deficiencies in progenitor cells of multiple hematopoietic lineages and defective megakaryocytopoiesis in mice lacking the thrombopoietic receptor c-Mpl. Blood. 1996;87:2162–70.

    CAS  PubMed  Google Scholar 

  44. Lebois M, Dowling MR, Gangatirkar P, Hodgkin PD, Kile BT, Alexander WS, et al. Regulation of platelet lifespan in the presence and absence of thrombopoietin signaling. J Thromb Haemost. 2016;14:1882–7.

    CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Prof Benjamin Kile for generously providing Bcl-xPlt20 mice and A/Prof Marie-Liesse Asselin-Labat for kindly sharing cell lines. We thank Janelle Lochland, Dr Amanda Au, Nicole Lynch, Stephanie Bound, Keti Stoev, Rebekah Meeny and Shannon Oliver for outstanding assistance and Prof Doug Hilton for helpful discussions. Ann Officer facilitated data collection for the Thoracic Malignancies Cohort Study and Him Ahmad assisted with patient pathology data collation. This work was supported by Australian National Health and Medical Research Council Project, Ideas, and Program Grants (1079250 ECJ, 1113577, 1122783 APN, 1186575 MJH, 1159658 MJH), Fellowships (1058344 WSA, 1156095 MJH), an Independent Research Institutes Infrastructure Support Scheme Grant (9000220), and a Victorian State Government Operational Infrastructure Support Grant. ECJ is the recipient of a fellowship from the Lorenzo and Pamela Galli Charitable Trust. SRH is the recipient of an Australian Postgraduate Award from the University of Melbourne. KDS is supported by a Victorian Cancer Agency Mid-Career Fellowship (MCRF18003) and the Peter and Julie Alston Centenary Fellowship. The generation of the NSG Mpl−/− mice was supported by the Australian Phenomics Network, the Leukemia and Lymphoma Society (LLS SCOR 7001-13 MJH), and the Australian Government through the National Collaborative Research Infrastructure Strategy program.

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Authors

Contributions

SRH designed and performed research, analysed data, prepared figures, and wrote the paper. ECJ conceived, designed and supervised research, and wrote the paper. MA, AT, AK, AJK, MJH, JC, PG and APN conducted experiments and analysed data. KDS provided the inducible ADC model. WSA and KDS designed research and revised the paper. KDS, WSA, BS, MA, AT and APN contributed to intellectual discussions of the data. All authors reviewed the paper.

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Correspondence to Emma C. Josefsson.

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Conflict of interest

BJS has served on advisory boards for AstraZeneca, Roche/Genentech, Pfizer, Merck, Bristol Myers Squibb, Novartis, Gritstone Oncology, Loxo Oncology.

Ethical approval

Analysis of human clinical data was performed according to National Statement on Ethical Conduct in Human Research and was approved by the Peter MacCallum Cancer Centre ethics committee (approval no.17/136R). All mice were handled in accordance with approved protocols (Walter and Eliza Hall Institute Animal Ethics Committee).

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Hyslop, S.R., Alexander, M., Thai, A.A. et al. Targeting platelets for improved outcome in KRAS-driven lung adenocarcinoma. Oncogene 39, 5177–5186 (2020). https://doi.org/10.1038/s41388-020-1357-6

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