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bioRxiv · 10.1101/2025.09.13.676018

Interspecies Functional Divergence: The Microbiome's Role in FLOT Response e Across Gastric Cancer Subtypes

Abstract

BackgroundGastric adenocarcinoma exhibits marked molecular and histological heterogeneity, which is reflected in distinct patterns of progression, therapeutic response, and prognosis. Although the FLOT regimen (5-fluorouracil, leucovorin, oxaliplatin, and docetaxel) represents the current standard for perioperative chemotherapy, its systemic effects on the tumor microenvironment, including the associated bacterial microbiome and host gene expression, remain poorly understood. MethodsThis study investigated the effects of FLOT on the functional and ecological structure of intestinal and diffuse subtype gastric tumors by assessing its simultaneous influence on the human transcriptome, the bacterial transcriptome, and inter-kingdom interactions. We analyzed 55 tumor samples (37 intestinal subtype; 18 diffuse subtype) and explored potential genetic-functional interactions between the bacterial microbiome and the human genome. ResultsThe results reveal a highly specific functional pattern in the diffuse subtype, absent in the intestinal subtype, demonstrating a unique ecological-transcriptional plasticity mediated by the microbiota under chemotherapeutic pressure. The interaction network was dominated by high-magnitude positive correlations. Notably, the bacterial gene leuS showed a robust association with the human gene HCN1 and processes such as potassium ion transmembrane transport, membrane depolarization, and regulation of postsynaptic membrane potential, indicating a coordinated activation of ion channels and neuroepithelial circuits. Bacterial species including Bacteroides uniformis, Faecalibacterium prausnitzii, Butyrivibrio crossotus, Prevotella copri, and Simiaoa sunii also converged functionally on HCN1. Additionally, bacterial genes mfd, nifJ, secY, rplF, and tet(Q) were associated with pathways related to cell adhesion, epithelial proliferation, membrane potential control, and synaptic transduction. DiscussionIntegrative analysis reveals that the FLOT regimen acts as a systemic remodeler of the gastric tumor microenvironment, exerting distinct effects according to the histological subtype. While the intestinal subtype responds more aligned with the cytotoxic goals of chemotherapy, the diffuse subtype exhibits a functional plasticity that favors the emergence of adaptive and possibly pro-tumoral phenotypes. We propose a mechanistic model where in chemotherapy selectively reshapes the microbial ecosystem, which in turn modulates host functional circuits, directly influencing tumor behavior. These findings open perspectives for combined therapeutic strategies that include targeted modulation of the microbiome as an adjuvant to chemotherapy.

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BibTeXRIS

Sarquis, D. P., Santos da Silva, V. C., Avelar, D. d. S., da Silva, J. M. C., Mourao, R. M. d. S., Araujo, T. M. T., Pereira, D., Araujo, P. S. d., Ishak, G., Moraes Casseb, S. M., Demachki, S., Barra, W. F., Moreira, F. C., Burbano, R. M. R., Assumpcao, P. P.. 2025-09-17. Interspecies Functional Divergence: The Microbiome's Role in FLOT Response e Across Gastric Cancer Subtypes. https://doi.org/10.1101/2025.09.13.676018

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