Deuterium Concentration as a Dual Regulator: Depletion Suppresses While Enrichment Amplifies Oncogenic Hallmarks in Lung Adenocarcinoma

Background: Deuterium abundance has emerged as a metabolic regulator of oncogenic transcriptional programs, yet its impact on cancer gene networks remains incompletely defined. Methods: We profiled A549 lung adenocarcinoma cells cultured under graded deuterium concentrations (40, 80, 150, 300 ppm) using NanoString nCounter technology. Expression data were subjected to multistep filtering, symbolic trajectory encoding, density‑based spatial clustering (DBSCAN) to identify sentinel outliers, and Gaussian mixture modeling (GMM‑6) to resolve coherent transcriptional modules. Results: DBSCAN analysis identified 11 extreme responder genes that define specific vulnerabilities under deuterium depletion, including suppression of multidrug resistance ( ABCB1 , −42% at 80 ppm), proliferative signaling ( FGFR4 , −19%), and transcriptional amplification ( MYCN , −24%). In contrast, enrichment at 300 ppm drove a global activation of oncogenic pathways (mean +44%), particularly amplifying tumor‑promoting inflammation ( IL6 , TGFBR2 ) and invasion/metastasis ( MMP9 ). GMM‑6 clustering of the remaining core network resolved six functional modules, revealing that depletion selectively disengages the high‑energy programs required for phenotypic plasticity (Cluster 5: TGFB1 , S100A4 ) while leaving basal survival reserves (Cluster 6: BIRC5 , RET ) relatively intact. Conclusions: These findings establish deuterium concentration as a dual regulator of oncogenic transcription. Enrichment fuels a hyper‑proliferative, invasive state, whereas moderate depletion exposes stress‑responsive vulnerabilities by selectively suppressing drug resistance, growth factor signaling, and invasive plasticity in the A549 lung adenocarcinoma model.