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Molecular Diagnostics

Unveiling the origin and functions of diagnostic circulating microRNAs in lung cancer

British Journal of Cancer volume 132, pages 947–956 (2025)Cite this article

Subjects

Abstract

Background

Circulating microRNAs (c-miRs) were shown to be effective biomarkers for lung cancer early detection. However, the understanding of c-miRs origin and their biological functions still remains elusive.

Methods

We analysed miRNA expression in a large panel of lung cancer (LC) and hematopoietic cell lines (N = 252; CCLE database) coupled with c-miR profile of a large cohort of serum samples (N = 975), from high-risk subjects underwent annual LD-CT for 5 years. Furthermore, we examined intracellular and extracellular miR-29a-3p/223-3p expression profile in lung adenocarcinoma (LUAD) tissues, in matched serum samples and in LC and stromal cell lines. Lastly, through the modulation of expression of selected c-miRs by using mimic (OE) or antisense microRNA (KD), we explored their impact on lung cancer transcriptome and cancer and immune phenotypes.

Results

Here, we investigated the origin of an extensively validated 13 c-miRs signature diagnostics for asymptomatic lung cancer (LC) in high-risk subjects (smokers, >20 packs/y; >50 y old). Overall, we found a mixed origin of these c-miRs, originating both from tumour cells and the tumour microenvironment (TME). Intriguingly, we revealed that circulating miR-29a-3p and miR-223-3p are abundantly released from LC epithelial cells and immune cells, respectively. In particular, we found that miR-223-3p triggered several lung cancer related phenotypes such as invasion, migration and tumour-promoting inflammation.

Conclusions

Our study highlights a mixed tumour epithelial and stroma-associated origin of LC c-miRs with new evidences on the multifaceted role of miR-223-3p in LC pathogenesis and immune modulation.

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Fig. 1: Expression analysis of the 13 c-miRs LC diagnostic signature (miR-Test) across cell lines, patient samples and tissue compartments.
Fig. 2: Transcriptomic analysis of miR-29a-3p and miR-223-3p in lung cancer cells.
Fig. 3: Analysis of the regulatory roles of miR-29a-3p and miR-223-3p in lung cancer cells and T-cells activation.
Fig. 4: miR-223-3p shapes cytokine expression, immune cell composition, and clinical outcomes in LUAD patients.

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Data availability

Raw Data of this study (Affymetrix data) can be found in Gene Expression Omnibus database with the following accession number: GSE271130.

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Acknowledgements

We would like to thank Teresa Nittoli for helping us in experiments setting up as well as Orazio Palumbo for Affymetrix experiments.

Funding

The research leading to these results has received funding from the Italian Ministry of Health [GR-2016-02363975 to FB; RF-2021-12372433 to F.B.; GR-2019-12370460 to TC] and from Fondazione AIRC per la Ricerca sul Cancro ETS [IG 2019—ID. 22827 project to FB; IG2024 – ID. 30689]. FM was a recipient of a fellowship (Italy Pre-Doc) from AIRC ETS (ID. 28243). MKA is a recipient of a fellowship (Italy Pre-Doc) from AIRC ETS (ID. 29714).

Author information

Author notes

  1. Tommaso Colangelo

    Present address: Department of Medical and Surgical Sciences, University of Foggia, Foggia, Italy

  2. Paolo Graziano

    Present address: Department of Radiological, Oncological and Pathological Sciences, Sapienza University of Rome, Roma, Italy

  3. These authors contributed equally: Tommaso Colangelo, Francesco Mazzarelli, Roberto Cuttano.

Authors and Affiliations

  1. Unit of Cancer Biomarkers, Fondazione IRCCS Casa Sollievo della Sofferenza, San Giovanni Rotondo, Italy

    Tommaso Colangelo, Francesco Mazzarelli, Roberto Cuttano, Elisa Dama, Valentina Melocchi, Miriam Kuku Afanga, Rosa Maria Perrone & Fabrizio Bianchi

  2. Unit of Pathology, Fondazione IRCCS Casa Sollievo della Sofferenza, San Giovanni Rotondo, Italy

    Paolo Graziano

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Contributions

Conceptualisation: FB; methodology: TC, FM, RC, MKA, RMP, VM, ED, PG, FB; investigation: TC, FM, RC, RMP, VM, FB; visualisation: TC, FM, VM, FB; supervision: FB; writing (original draft): FB; writing (review and editing): TC, RC, FB.

Corresponding author

Correspondence to Fabrizio Bianchi.

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Competing interests

The study funders had no role in the design of the study, the collection, analysis and interpretation of the data, the writing of the manuscript and the decision to submit the manuscript for publication. All the authors declare no conflict of interest.

Ethics approval and consent to participate

Study was approved by internal IRB at IRCCS Casa Sollievo della Sofferenza Hospital (protocol name: BIOPOLMONE v1.0_08 Giu 16). Patients signed an informed consent in accordance with the Helsinki’s declaration.

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Colangelo, T., Mazzarelli, F., Cuttano, R. et al. Unveiling the origin and functions of diagnostic circulating microRNAs in lung cancer. Br J Cancer 132, 947–956 (2025). https://doi.org/10.1038/s41416-025-02982-x

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