Integrative transcriptomic and cistromic analyses reveal endothelial mechanotransduction–clock coupling and TEAD-dependent regulation of CRY1 and CLOCK in pulmonary arterial hypertension

Pulmonary arterial hypertension (PAH) is characterized by vascular remodeling within an abnormal mechanical microenvironment, but the relationship between mechanotransduction and clock-related transcription in the pulmonary vasculature remains poorly defined. We integrated three PAH lung transcriptomic cohorts, donor-aware endothelial pseudobulk, independent pulmonary vascular endothelial datasets, a high-resolution single-cell atlas, digital spatial transcriptomics, endothelial YAP/TAZ perturbation transcriptomics, and YAP/TAZ/TEAD1 ChIP-seq, followed by validation in human pulmonary artery endothelial cells (HPAECs). In GSE117261, YAP/TAZ–TEAD, FAK/SRC, and overall mechanotransduction scores correlated positively with the clock-related score. Donor-level endothelial analyses supported positive coupling in GSE169471 and GSE210248, whereas vascular-state and spatial analyses demonstrated marked context dependence. In human umbilical vein endothelial cells (HUVECs), constitutively active YAP or TAZ increased CRY1 and CLOCK, while TEAD-binding-deficient mutants attenuated these responses. Cistromic reanalysis identified convergent YAP, TAZ, and TEAD1 occupancy at candidate CRY1/CLOCK regulatory regions with nearby TEAD motifs and active/open-chromatin support. In HPAECs, YAP-S127A and TAZ-S89A increased CRY1/CLOCK mRNA and protein, whereas corresponding TEAD-binding-deficient mutants markedly blunted these effects. These findings indicate that mechanotransduction is coupled to clock-related transcription predominantly in endothelial contexts in human PAH, with this relationship varying across cellular and spatial states. They further support a requirement for an intact interface between YAP/TAZ and TEAD in the regulation of CRY1 and CLOCK, providing a candidate molecular link between mechanical signaling and clock-related transcriptional remodeling.