Tissue MicroRNA Expression as a Predictor of Response to Immunotherapy in NSCLC Patients

Anna Grenda; Paweł Krawczyk; Justyna Błach; Izabela Chmielewska; Tomasz Kubiatowski; Stanisław Kieszko; Kamila Wojas-Krawczyk; Tomasz Kucharczyk; Bożena Jarosz; Iwona Paśnik; Małgorzata Borowiec-Bar; Małgorzata Frąk; Robert Kieszko; Michał Szczyrek; Katarzyna Reszka; Kinga Krukowska; Agnieszka Kolak; Sławomir Mańdziuk; Dariusz Kowalski; Marek Sawicki; Daria Świniuch; Elżbieta Starosławska; Rodryg Ramlau; Justyna Szumiło; Maciej Krzakowski; Janusz Milanowski

doi:10.3389/fonc.2020.563613

Tissue MicroRNA Expression as a Predictor of Response to Immunotherapy in NSCLC Patients

Front Oncol. 2021 Feb 8:10:563613. doi: 10.3389/fonc.2020.563613. eCollection 2020.

Authors

Anna Grenda¹, Paweł Krawczyk¹, Justyna Błach¹, Izabela Chmielewska¹, Tomasz Kubiatowski², Stanisław Kieszko², Kamila Wojas-Krawczyk¹, Tomasz Kucharczyk¹, Bożena Jarosz³, Iwona Paśnik⁴, Małgorzata Borowiec-Bar¹, Małgorzata Frąk¹, Robert Kieszko¹, Michał Szczyrek¹, Katarzyna Reszka⁵, Kinga Krukowska⁵, Agnieszka Kolak⁶, Sławomir Mańdziuk⁶, Dariusz Kowalski⁷, Marek Sawicki⁸, Daria Świniuch⁹, Elżbieta Starosławska², Rodryg Ramlau⁹, Justyna Szumiło⁴, Maciej Krzakowski⁷, Janusz Milanowski¹

Affiliations

¹ Department of Pneumonology, Oncology and Allergology, Medical University of Lublin, Lublin, Poland.
² Department of Clinical Oncology, Saint John of Dukla Oncology Centre of the Lublin Region, Lublin, Poland.
³ Department of Neurosurgery and Paediatric Neurosurgery, Medical University of Lublin, Lublin, Poland.
⁴ Department of Clinical Pathomorphology, Medical University of Lublin, Lublin, Poland.
⁵ Genetics and Immunology Institute of Lublin, Genim LLC, Lublin, Poland.
⁶ Department of Clinical Oncology and Chemotherapy, Medical University of Lublin, Lublin, Poland.
⁷ Department of Lung and Chest Cancer, The Maria Sklodowska-Curie National Research Institute of Oncology in Warsaw, Warsaw, Poland.
⁸ Department of Thoracic Surgery, Medical University of Lublin, Lublin, Poland.
⁹ Department of Oncology, Poznan University of Medical Sciences, Poznań, Poland.

Abstract

Introduction: Expression of PD-L1 protein on tumor cells, which is so far the only validated predictive factor for immunotherapy, is regulated by epigenetic and genetic factors. Among the most important ones that regulate gene expression are microRNAs.

Materials and methods: The study included 60 patients with NSCLC who underwent first or second line immunotherapy with pembrolizumab or nivolumab. FFPE materials were collected before the start of immunotherapy. We examined relative expression of microRNAs (miR-141, miR-200a, miR-200b, miR-200c, miR-429, miR-508-3p, miR-1184, miR-1255a) and PD-L1 mRNA expression. Copy number variation (CNV) of PD-L1 gene by qPCR and FISH methods were assessed. Two single nucleotide polymorphisms (SNPs) in promoter region of PD-L1 gene (rs822335 and rs822336) were examined. Expression of PD-L1 protein on tumor cells was assessed by immunohistochemistry (IHC). The response rate to immunotherapy and progression free survival (PFS) measured in weeks and overall survival (OS) measured in months from the start of immunotherapy were evaluated.

Results: Response to immunotherapy was observed in nine patients (15%, including one complete response), disease stabilization in 22 patients (36.7%), and progression in 29 patients (48.3%). Significantly higher (p=0.015) expression of miR-200b and significantly lower (p=0.043) expression of miR-429 were observed in responders compared to patients who did not respond to immunotherapy. The median PFS in the whole group of patients was 16 weeks, and the median OS was 10.5 month. In univariate analysis, the median PFS was significantly higher in patients with high miR-200b expression (HR=0.4253, 95%CI: 0.1737-1.0417, p=0.05) and high miR-508 expression (HR=0.4401, 95%CI: 0.1903-1.0178, p=0.05) and with low expression of miR-429 (HR=0.1288, 95%CI: 0.01727-0.9606, p=0.0456) compared to patients with low and high expression of these molecules, respectively. The median OS was higher in patients with low expression of miR-429 (HR=0,6288, 95%CI: 0,3053-1,2949, p=0.06) compared with patients with high expression of this microRNA. In multivariate analysis, we found that patients with PD-L1 expression on ≥1% of tumor cells compared to patients without PD-L1 expression on cancer cells had a significantly lower risk of progression (HR=0.3857, 95%CI: 0.1612-0.9226, p=0.0323) and death (HR=0.377, 95%CI: 0.1636-0.8688, p=0.022).

Conclusion: The miR-200b and miR-429 molecules in tumor cells seem to have greatest impact on the effectiveness of immunotherapy in NSCLC patients.

Keywords: PD-L1; SNP; copy number variation; immunotherapy; microRNA; non-small cell lung cancer.

Copyright © 2021 Grenda, Krawczyk, Błach, Chmielewska, Kubiatowski, Kieszko, Wojas-Krawczyk, Kucharczyk, Jarosz, Paśnik, Borowiec-Bar, Frąk, Kieszko, Szczyrek, Reszka, Krukowska, Kolak, Mańdziuk, Kowalski, Sawicki, Świniuch, Starosławska, Ramlau, Szumiło, Krzakowski and Milanowski.