Phenotypic profle of monocyte-macrophage lineage cells as a function of respiratory epithelium status

The mechanism of the relationship between pretumor changes in the bronchial respiratory epithelium and the risk of progression of non-small cell lung cancer (NSCLC) remains unclear. It has been suggested that the relationship between reactive changes in the bronchial mucosa and NSCLC progression may be caused by the functional status of monocytic-macrophage cells as important participants in infammation, which determines both the risk of premalignant changes in the epithelium and malignant progression.
The purpose of the study was to investigate the phenotypic profle of peripheral blood monocytes and macrophages induced from monocytes in vitro depending on the state of respiratory epithelium in NSCLC patients.
Material and Methods. The study included 39 patients with newly diagnosed NSCLC. Based on the morphological examination of small bronchi taken at the distance of 3–5 cm from the tumor, patients were divided into the following groups depending on the type of pretumor changes: no pretumor changes (n=6), isolated basal cell hyperplasia (BCH) (n=13), combination of BCH and squamous metaplasia (SM) (n=3), combination of unchanged epithelium and focal BCH (n=17). The phenotypic features of peripheral blood monocytes and in vitro-induced macrophages were assessed before treatment using fow cytometry.
Results. The state of the respiratory epithelium in NSCLC patients prior to the start of anticancer treatment was associated with the phenotypic features of peripheral blood monocytes, but not with the profle of macrophages induced from them. Distortion of the response of induced macrophages to the polarizing stimuli was observed in NSCLC patients: the cultured cells responded to both M1 and M2 inducers (LPS and IL-4, respectively) with a phenotype shift to M2, while the CD206 marker expression varied depending on the presence and type of pretumor changes.
Conclusion. The phenotypic profle of peripheral blood monocytes was associated with the state of the respiratory epithelium in NSCLC patients before anti-tumor treatment, but not with the phenotypic features of induced macrophages.

1. Majem M., Juan O., Insa A., Reguart N., Trigo J.M., Carcereny E., García-Campelo R., García Y., Guirado M., Provencio M. SEOM clinical guidelines for the treatment of non-small cell lung cancer (2018). Clin Transl Oncol. 2019; 21(1): 3–17. doi: 10.1007/s12094-018-1978-1.

2. Duma N., Santana-Davila R., Molina J.R. Non-Small Cell Lung Cancer: Epidemiology, Screening, Diagnosis, and Treatment. Mayo Clin Proc. 2019; 94(8): 1623–40. doi: 10.1016/j.mayocp.2019.01.013.

3. Pankova O.V., Denisov E.V., Ponomaryova A.A., Gerashchenko T.S., Tuzikov S.A., Perelmuter V.M. Recurrence of squamous cell lung carcinoma is associated with the co-presence of reactive lesions in tumor-adjacent bronchial epithelium. Tumour Biol. 2016; 37(3): 3599–607. doi: 10.1007/s13277-015-4196-2.

4. Yao Y., Xu X.H., Jin L. Macrophage Polarization in Physiological and Pathological Pregnancy. Front Immunol. 2019; 10: 792. doi: 10.3389/fmmu.2019.00792.

5. 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(1): 3. doi: 10.1186/s12865-018-0241-4.

6. Yao Y., Xu X.H., Jin L. Macrophage Polarization in Physiological and Pathological Pregnancy. Front Immunol. 2019; 10: 792. doi: 10.3389/fmmu.2019.00792.

7. Cassetta L., Pollard J.W. Targeting macrophages: therapeutic approaches in cancer. Nat Rev Drug Discov. 2018; 17(12): 887–904. doi: 10.1038/nrd.2018.169.

8. Poschke I., Mao Y., Adamson L., Salazar-Onfray F., Masucci G., Kiessling R. Myeloid-derived suppressor cells impair the quality of dendritic cell vaccines. Cancer Immunol Immunother. 2012; 61(6): 827–38. doi: 10.1007/s00262-011-1143-y.

9. Cao X., Yakala G.K., van den Hil F.E., Cochrane A., Mummery C.L., Orlova V.V. Diferentiation and Functional Comparison of Monocytes and Macrophages from hiPSCs with Peripheral Blood Derivatives. Stem Cell Reports. 2019; 12(6): 1282–97. doi: 10.1016/j.stemcr.2019.05.003.

10. Chen H.J., Li Yim A.Y.F., Griffth G.R., de Jonge W.J., Mannens M.M.A.M., Ferrero E., Henneman P., de Winther M.P.J. Meta-Analysis of in vitro-Diferentiated Macrophages Identifes Transcriptomic Signatures That Classify Disease Macrophages in vivo. Front Immunol. 2019; 10: 2887. doi: 10.3389/fmmu.2019.02887.

11. Foulds G.A., Vadakekolathu J., Abdel-Fatah T.M.A., Nagarajan D., Reeder S., Johnson C., Hood S., Moseley P.M., Chan S.Y.T., Pockley A.G., Rutella S., McArdle S.E.B. Immune-Phenotyping and Transcriptomic Profling of Peripheral Blood Mononuclear Cells From Patients With Breast Cancer: Identifcation of a 3 Gene Signature Which Predicts Relapse of Triple Negative Breast Cancer. Front Immunol. 2018; 9: 2028. doi: 10.3389/fmmu.2018.02028.

12. Gurvich O.L., Puttonen K.A., Bailey A., Kailaanmäki A., Skirdenko V., Sivonen M., Pietikäinen S., Parker N.R., Ylä-Herttuala S., Kekarainen T. Transcriptomics uncovers substantial variability associated with alterations in manufacturing processes of macrophage cell therapy products. Scientifc reports. 2020; 10(1). doi: 10.1038/s41598-020-70967-2.

13. Hamidzadeh K., Belew A.T., El-Sayed N.M., Mosser D.M. The transition of M-CSF-derived human macrophages to a growth-promoting phenotype. Blood Adv. 2020; 4(21): 5460–72. doi: 10.1182/bloodadvances.2020002683.

14. Jin X., Kruth H.S. Culture of Macrophage Colony-stimulating Factor Diferentiated Human Monocyte-derived Macrophages. J Vis Exp. 2016; (112). doi: 10.3791/54244.

15. Lukic A., Larssen P., Fauland A., Samuelsson B., Wheelock C.E., Gabrielsson S., Radmark O. GM-CSF- and M-CSF-primed macrophages present similar resolving but distinct infammatory lipid mediator signatures. FASEB J. 2017; 31(10): 4370–81. doi: 10.1096/fj.201700319R.

16. Beane J.E., Mazzilli S.A., Campbell J.D., Duclos G., Krysan K., Moy C., Perdomo C., Schaffer M., Liu G., Zhang S., Liu H., Vick J., Dhillon S.S., Platero S.J., Dubinett S.M., Stevenson C., Reid M.E., Lenburg M.E., Spira A.E. Molecular subtyping reveals immune alterations associated with progression of bronchial premalignant lesions. Nat Commun. 2019; 10(1): 1856. doi: 10.1038/s41467-019-09834-2.

17. Chen S., Yang J., Wei Y., Wei X. Epigenetic regulation of macrophages: from homeostasis maintenance to host defense. Cell Mol Immunol. 2020; 17(1): 36–49. doi: 10.1038/s41423-019-0315-0.

18. Larionova I., Kazakova E., Patysheva M., Kzhyshkowska J. Transcriptional, Epigenetic and Metabolic Programming of Tumor-Associated Macrophages. Cancers (Basel). 2020; 12(6): 1411. doi: 10.3390/cancers12061411.

19. Gabrilovich D.I., Ostrand-Rosenberg S., Bronte V. Coordinated regulation of myeloid cells by tumours. Nat Rev Immunol. 2012; 12(4): 253–68. doi: 10.1038/nri3175.

20. Larionova I., Tuguzbaeva G., Ponomaryova A., Stakheyeva M., Cherdyntseva N., Pavlov V., Choinzonov E., Kzhyshkowska J. Tumor-Associated Macrophages in Human Breast, Colorectal, Lung, Ovarian and Prostate Cancers. Front Oncol. 2020; 10. doi: 10.3389/fonc.2020.566511.

21. Orecchioni M., Ghosheh Y., Pramod A.B., Ley K. Macrophage Polarization: Diferent Gene Signatures in M1(LPS+) vs. Classically and M2(LPS-) vs. Alternatively Activated Macrophages. Frontiers in immunology. 2019; 10: 1084. doi: 10.3389/fmmu.2019.01084.