Abstract Cancer-associated fibroblasts (CAFs) display significant phenotypic heterogeneity driven by dynamic interactions within the tumor microenvironment. To investigate motility-driven CAF heterogeneity, we utilized a 3D culture system and the photoconvertible fluorescent protein Dendra2, to isolate ‘motile’ and ‘static’ CAF subpopulations. Transcriptomic analyses revealed that motile CAFs upregulated hypoxia- and glycolysis-related genes, which are downstream targets of HIF-1α. In line with these findings, HIF-1α activation enhanced CAF motility, while the knockdown of its downstream targets, ALDOA and LDHA, significantly reduced CAF motility. Notably, mesenchymal-like lung cancer cells enhanced both HIF-1α activity and motility in CAFs through secretory factors. Using mass spectrometry, we identified ceruloplasmin (CP) as a key secretory protein from these lung cancer cells, whose transcription was regulated by the EMT-inducing transcription factor ZEB1. Functionally, CP knockdown abolished the ability of lung cancer cells to promote CAF motility in vitro and to metastasize in vivo. Collectively, these findings demonstrate a novel mechanism where lung cancer cell-secreted CP facilitates HIF-1α signaling in CAFs, thereby driving their motility and promoting metastasis. Citation Format: Jihye Park, Sieun Lee, Jonathan M. Kurie, Young-Ho Ahn. Cancer cell-driven HIF-1α activation enhances the motility of cancer-associated fibroblasts in the lung tumor microenvironment abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 6039.
Park et al. (Fri,) studied this question.