NSCLC, comprising LUAD and LUSC, remains a leading cause of cancer mortality worldwide [20]. The tumor microenvironment significantly influences NSCLC progression, with M2 macrophages, a subtype of tumor-associated macrophages, playing a pivotal role in promoting immunosuppression and tumor growth [21, 22]. MRO Maestro, a gene implicated in various physiological processes, including the modulation of protein–protein interactions via HEAT repeat motifs, has shown potential relevance in the tumor microenvironment. However, its specific role and prognostic value in NSCLC, particularly in relation to M2 macrophages and immune infiltration, remain unclear. This study investigates the expression of MRO in NSCLC and its potential as a prognostic biomarker and therapeutic target.

Our analysis revealed significant expression of MRO in NSCLC, particularly in LUAD and LUSC. This differential expression suggests that MRO may play a unique role in the pathophysiology of these NSCLC subtypes. The strong correlation between MRO expression and M2 macrophage presence underscores its potential involvement in the immunosuppressive and tumor-promoting functions of these cells. M2 macrophages facilitate tumor progression by secreting anti-inflammatory cytokines, promoting tissue remodeling, and supporting angiogenesis [23]. MRO has emerged as a critical factor in M2 macrophage polarization, particularly within the contexts of cancer and inflammatory diseases. Studies suggest that M2 macrophages, which are associated with tumor progression and immune suppression, are modulated by a variety of genes, including MRO. This highlights the potential role of MRO in shaping the tumor microenvironment through its influence on macrophage behavior [24]. Thus, MRO could be a key regulator of these processes in the NSCLC tumor microenvironment.

Further supporting its role in the tumor microenvironment, the ESTIMATE algorithm demonstrated a positive correlation between MRO expression and stromal, immune, and ESTIMATE scores. Higher MRO expression is associated with increased immune cell infiltration and stromal content. Pathway enrichment analyses revealed significant associations between MRO expression and key immune pathways, including TNFα signaling via NFκB [25], inflammatory response [26], and IL6 JAK STAT3 signaling [27]. These pathways are critical for orchestrating immune responses and inflammation, suggesting that MRO may modulate these processes to influence tumor progression and immune evasion.

Building on these associations, our survival analyses highlighted the prognostic significance of MRO in NSCLC. High MRO expression was associated with poorer OS, indicating its potential as a prognostic biomarker. The correlation between high MRO levels and adverse outcomes may reflect its role in promoting an immunosuppressive tumor microenvironment, facilitating tumor growth and resistance to therapies. These findings suggest that MRO could be instrumental in shaping the tumor microenvironment in ways that promote tumor progression and reduce the efficacy of conventional therapies. In addition to its prognostic significance, our genomic analyses revealed significant correlations between MRO expression and key genomic instability markers, such as TMB, MATH, and MSI. Notably, MRO showed strong negative correlations with MATH and MSI, suggesting it may play a role in maintaining genomic stability [28, 29]. Additionally, mutation landscape analysis identified specific mutations associated with MRO expression, further elucidating its role in NSCLC. The differential mutation profiles between high and low MRO expression groups highlight the genetic complexity and heterogeneity of NSCLC.

The implications of our findings extend to potential therapeutic strategies. TIDE and Submap analyses suggested that MRO expression may influence the response to immunotherapy. Low MRO expression was associated with lower TIDE scores and potentially better responses to immunotherapy in LUSC patients, whereas no significant difference was observed in LUAD patients. This differential response highlights the need for further investigation into the context-specific roles of MRO in NSCLC subtypes. Targeting MRO in combination with existing therapies could enhance treatment efficacy by modulating the immune microenvironment and improving patient responses to immunotherapy.

In conclusion, our study underscores the multifaceted role of MRO in NSCLC, particularly its association with M2 macrophages, immune infiltration, and prognosis. MRO emerges as a promising biomarker for prognostication and a potential target for therapeutic intervention. Future studies should aim to validate these findings through experimental and clinical investigations, exploring the mechanistic underpinnings of MRO's role in NSCLC. Understanding how MRO modulates immune pathways and interacts with the tumor microenvironment will be crucial for developing targeted therapies that can improve patient outcomes in NSCLC.

This study has several limitations that warrant consideration. Firstly, our reliance on bioinformatics and publicly available datasets introduces potential biases, underscoring the need for experimental validation to confirm our findings. Additionally, the inherent heterogeneity of NSCLC and the influence of various molecular factors were not fully accounted for, which may restrict the generalizability of our conclusions. Furthermore, while computational tools such as ESTIMATE, TIDE, and SubMap offer valuable insights into tumor microenvironments, their inherent limitations and underlying assumptions may affect the interpretation of results. Therefore, we recognize that our findings should be viewed as preliminary and suggestive rather than definitive. In light of these limitations, we acknowledge the critical need for future research that integrates multi-omics data and performs functional assays. Such approaches would provide a more comprehensive understanding of the role of MRO in NSCLC and its potential as a therapeutic target. Validation through prospective experimental studies is essential to establish any causal relationships and further elucidate the mechanisms involved.