Lung Tissue Microbiome in NSCLC Patients - Metabarcoding Analysis Identifies Escherichia-Shigella as an Abundant Taxon

Sep 22, 2026

Non-small cell lung cancer (NSCLC) remains the leading cause of cancer-related mortality worldwide, and accumulating evidence links lung microbiome alterations to NSCLC development and progression. Findings have been inconsistent across studies due to heterogeneous biological materials and differing methods. This study characterized the lung tissue microbiome using paired tumor and adjacent tissue samples, applying 16S rRNA V3–V4 sequencing and QIIME2 bioinformatics. Tumors showed reduced microbial diversity and highlighted Escherichia-Shigella abundance as a candidate for further investigation.

LanguageEnglish

CategoryResearch & Report

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1. Introduction

Article Lung Tissue Microbiome in NSCLC Patients: Metabarcoding Analysis Identifies Escherichia-Shigella as an Abundant Taxon Piotr Machnicki 1, Karolina Czarnecka-Chrebelska 2, Jacek Kordiak 3, Krzysztof Lewandowski 3, Filip Bielec 1, Tomasz Płoszaj 4, Ewa Brzezia ´nska-Lasota 2 and Dorota Pastuszak-Lewandoska 1, * Academic Editors: Jeffrey A. Borgia and Maria Gazouli Received: 20 April 2026 Revised: 23 June 2026 Accepted: 26 June 2026 Published: 29 June 2026 Copyright: © 2026 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license. 1 Department of Microbiology and Laboratory Medical Immunology, Medical University of Lodz, 90-151 Lodz, Poland; [piotr.machnicki@umed.lodz.pl](piotr.machnicki@umed.lodz.pl) (P.M.); [filip.bielec@umed.lodz.pl](filip.bielec@umed.lodz.pl) (F.B.) 2 Department of Biomedicine and Genetics, Medical University of Lodz, Mazowiecka 5, 92-215 Lodz, Poland; [karolina.czarnecka@umed.lodz.pl](karolina.czarnecka@umed.lodz.pl) (K.C.-C.); [ewa.brzezianska@umed.lodz.pl](ewa.brzezianska@umed.lodz.pl) (E.B.-L.) 3 Department of Thoracic, General and Oncological Surgery, Medical University of Lodz, 90-151 Lodz, Poland; [jacek.kordiak@umed.lodz.pl](jacek.kordiak@umed.lodz.pl) (J.K.); [krzysztof.jerzy.lewandowski@umed.lodz.pl](krzysztof.jerzy.lewandowski@umed.lodz.pl) (K.L.) 4 Department of Clinical Genetics, Medical University of Lodz, 90-151 Lodz, Poland; tomasz.ploszaj@umed.lodz.pl * Correspondence: [dorota.pastuszak-lewandoska@umed.lodz.pl](dorota.pastuszak-lewandoska@umed.lodz.pl) Simple Summary A growing body of research suggests that microorganisms living in the human body may influence the development and progression of lung cancer, but existing findings are inconsistent due to differences in research designs. In this study, we aimed to address this issue by analyzing tumor tissue and adjacent lung tissue from the same patients, allowing for more accurate comparisons. Using advanced DNA sequencing technology, we investigated which bacteria are present in the lung tumor environment and how they interact. Our results showed that the microbial community is less diverse in cancerous tissue, and this simplification extends beyond the tumor itself. We also identified a potentially harmful, dominant genus Escherichia-Shigella, in both tumor and surrounding tissue, suggesting that it may serve as a useful marker for future research. These findings may help improve the understanding of the role of microorganisms in lung cancer and support the development of new diagnostic approaches. Abstract Background: Non-small cell lung cancer (NSCLC) remains the leading cause of cancerrelated mortality worldwide despite advances in diagnosis and treatment. Increasing evidence suggests that alterations in the lung microbiome may contribute to NSCLC development and progression; however, findings remain inconsistent due to heterogeneous biological materials and methodological differences among studies. Therefore, this study aimed to characterize the lung tissue microbiome in NSCLC using a paired tissue-based approach. Methods: Thirty-two patients with NSCLC were enrolled. For each patient, two samples were collected: primary tumor tissue and matched macroscopically unchanged adjacent lung tissue. The V3-V4 region of the 16S rRNA gene was amplified and sequenced, followed by bioinformatic analysis using the QIIME2 pipeline. Results: Tumor tissues demonstrated lower alpha (Shannon H = 9.60, q = 0.001) and beta (Jaccard pseudo-F = 1.26, q = 0.015) diversity compared with adjacent controls, indicating reduced microbial complexity within the tumor microenvironment. Escherichia-Shigella was the most abundant detected genus (~12%) in both groups, although without a statistically significant difference. Analysis of microbiome variation in relation to spatial distance between sample

FAQs

Why is a paired tissue-based approach used in this NSCLC study?

Paired sampling of primary tumor tissue and matched macroscopically unchanged adjacent lung tissue reduces uncontrolled differences and enables more accurate comparisons of microbiome features across the same patients.

How was the lung tissue microbiome analyzed?

The study amplified and sequenced the V3–V4 region of the 16S rRNA gene, followed by bioinformatic analysis using the QIIME2 pipeline.

What microbiome differences were observed between tumor and adjacent tissues?

Tumor tissues showed lower alpha and beta diversity, indicating reduced microbial complexity in the tumor microenvironment compared with adjacent controls.

Sep 22, 2026

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