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Tumor Microenvironment Modulates Lineage Plasticity in Lung Squamous Cell Carcinoma

Fujibayashi, Y.;Ogawa, H.;Li, Q.;Navab, R.;Koga, T.;Inoue, Y.;Pham, N.;Hinokuma, H.;Bernards, N.;Sakane, T.;Matsumura, K.;Hiraishi, Y.;Yokote, F.;Yanagihara, T.;Aoi, T.;Maniwa, Y.;Radulovich, N.;Tsao, M.;Yasufuku, K.

2026-06-23 Cancer Biology
10.64898/2026.06.22.733895 bioRxiv
Show abstract

Lung squamous cell carcinoma (LUSC) is the second most common type of lung cancer, yet therapeutic options remain limited. A deeper understanding of its biology and molecular pathogenesis is essential for developing new treatment strategies. Here, we investigated the mechanisms of phenotypic plasticity in LUSC by comparing organoid-derived orthotopic lung models (ODOLs) and subcutaneous xenograft models (ODXs). ODXs showed greater tumor growth, squamous differentiation, and extracellular matrix (ECM) organization compared to ODOLs. Transcriptomic analyses revealed upregulation of multiple HIF1 and SOX2 target genes together with enhanced hypoxia signaling in ODXs. CRISPR/Cas9-mediated HIF1-knockout ODXs showed reduced SOX2 expression, tumor growth, and ECM organization, whereas SOX2-knockout ODXs reduced tumor growth without affecting HIF1 and ECM organization. These results indicate that HIF1 regulates squamous lineage maintenance through SOX2 and ECM remodeling. Spatial transcriptomics revealed enrichment of basal cell-like and squamous-differentiated tumor states in ODXs, whereas ODOLs displayed less differentiated phenotypes. These findings identify the tumor microenvironment as a critical determinant of lineage plasticity in LUSC and provide mechanistic insight into how hypoxia shapes tumor differentiation.

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