Genetic Alterations Affect Immune Contexture of Non-Small Cell Lung Cancer: Ukrainian Study

Denys Kozakov^1,2*Nazarii Kobyliak^1,3Sofiia Livshun¹Oleksii Seleznov^1,4Olena Koshyk^1,4Alina Matvieieva¹Yaroslav Shparyk⁵Oleksii Kolesnik^6,7Yuliia Moskalenko⁸Oleksandr Vynnychenko⁸Roman Moskalenko⁸Serhii Kropyvko²Anna Khmel⁹Bogdana Shkarupii^10,11Oksana Sulaieva^1,12,4

• ¹Medical Laboratory CSD, Kyiv, Ukraine
• ²Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, Kyiv, Kiev Oblast, Ukraine
• ³Bogomolets National Medical University, Kyiv, Ukraine
• ⁴Kyiv Medical University, Kyiv, Ukraine
• ⁵Lviv Regional Oncology Medical Diagnostic Centre, Lviv, Ukraine
• ⁶International European University, Киев, Ukraine
• ⁷Denys Clinics, Kyiv, Ukraine
• ⁸Sumy State University, Sumy, Ukraine
• ⁹Specialized Mammalogical Center, Kyiv, Ukraine
• ¹⁰Audubon Bioscience, Kyiv, Ukraine
• ¹¹Ukrainian Association of Research Biobanks, Kyiv, Ukraine
• ¹²Ukrainian Association for Precision Medicine, Kyiv, Ukraine

Although the role of various genetic alterations was highlighted among factors affecting the response to immunotherapy in non-small cell lung cancer (NSCLC), the relations between oncogenic driver variants and changes in the cancer immunity cycle are still unclear.The study aimed to discover the links between the molecular and immune context of NSCLC.Materials and methods: This cohort study included 254 cases of NSCLC (193 Lung Adenocarcinomas (LUAD; 76%), and 61 squamous cell carcinomas (SCC; 24%), with pathology reports and next-generation sequencing (NGS) data available. First, the rate and spectrum of genetic alterations were assessed in the Ukrainian cohort. Second, we uncovered the relationship between the oncogenic driver mutations and PD-L1 expression in NSCLC. Finally, T-cytotoxic lymphocytes (CD8 + ) and tumor-associated macrophages (CD163 + ) were evaluated in samples with EGFR and KRAS mutations, ALK rearrangements and LUAD with no genetic findings. Immune desert, immune excluded and inflamed types of tumor immune microenvironment (TME) were defined according to the cancer immunity cycle.Results: More than half (52%) of the observed NSCLC cases harbored single (48.03%) or concomitant (3.94%) genetic alterations in oncogenes. The Ukrainian cohort demonstrated a high rate of EGFR (18,5%) and ALK rearrangements (9.4%) with a relatively moderate frequency of KRAS mutations (16.9%). NSCLC tumors with alterations in EGFR and ALK demonstrated a high incidence of PD-L1 expression and specific immune contexture. The number of CD8 + cells varied significantly between oncogene-driven and wild-type LUAD (P=0.019). Non-oncogene-addicted NSCLC demonstrated the prevalence of Inflamed TME rich in CD163+ macrophages. In contrast, over half of EGFR mutant LUAD cases possessed immune desert TME type, while ALK-rearranged and KRAS mutant NSCLC showed mostly immune excluded TME.The high rate of PD-L1 expression in NSCLC driven by EGFR and ALK alterations was accompanied by a prevalence of low immunogenicity with a shift toward ID TME in EGFR mutant tumors and IE TME in ALK-rearranged and KRAS mutant NSCLC. Further discovery of mechanisms affecting tumor immune contexture is needed for tailoring patient management in line with particular mechanisms of immune evasion.