Wong, H. C. | Search bioRxiv

bioRxiv2025

Natural killer cell TGF- signaling regulates senolytic activity and vascular patterning in the postnatal lung

BackgroundBronchopulmonary dysplasia (BPD) is a disease of neonatal lung development that is linked to impaired pulmonary vascularization, dysregulated transforming growth factor-{beta} (TGF-{beta}) signaling and the accumulation of senescent cells. Despite the established role for TGF-{beta} signaling in promoting vascular remodeling and suppressing the senolytic activity of natural killer (NK) cells, the contribution of NK cell TGF-{beta} signaling to postnatal lung patterning and the pathogenesis of BPD remains unclear. MethodsMice bearing an NK cell-selective deletion of the type-II TGF-{beta} receptor (Tgfbr2NK-/-) were analyzed for vascular and alveolar structure, lung NK cell infiltration, senescence markers and lung function testing across neonatal and adult timepoints. Single-cell RNA sequencing of lung tissue from both neonatal mice and human infants with BPD was performed. The effect of enhanced NK cell activity in a hyperoxia-induced model of BPD was assessed in Tgfbr2NK-/-neonates, as well as pharmacologically, using the TGF-{beta} ligand trap/IL-15 superagonist, HCW9218. ResultsNeonatal Tgfbr2NK-/- mice exhibited a baseline reduction in distal arteriolar density, impaired alveolarization, and sex-specific deficits in long-term lung function. Single-cell RNA sequencing identified the excessive clearance of senescent endothelial cells by TGF-{beta} insensitive NK cells in the lungs of Tgfbr2NK-/- neonates, which served as a contributor of the BPD-like phenotype observed in naive animals. Tgfbr2NK-/- mice were protected from impaired lung development in the hyperoxia model. Sequencing from lung tissue from infants with BPD confirmed excessive TGF-{beta} signaling and cytotoxic impairment in NK cells. Treatment with HCW9218 prevented senescent cell accumulation and rescued lung development in the hyperoxia mouse model. ConclusionsThese findings identify TGF-{beta} as a tunable regulator of NK cell senolytic activity that is essential to normal postnatal lung development. Excessive NK cell TGF-{beta} signaling contributes to impaired lung development following exposure to neonatal hyperoxia and may serve as a viable therapeutic target for human BPD.

developmental biology↗

bioRxiv2025

Fluidic Programmable Gravi-maze Array for High Throughput Multiorgan Drug Testing

The high attrition rate of drug candidates in clinical trials underscores the urgent need for more predictive preclinical models that accurately replicate human physiology. Traditional 2D cell cultures and animal models often fail to predict human responses due to their limited physiological relevance, particularly for biologics and immunotherapies involving complex multicellular and cross-organ interactions. This highlights the need for modeling and measurements of multiorgan interactions at higher throughput, prompting the development of multiorgan-on-a-plate platforms. Here, we present OrganRX, a modular, gravity-driven recirculation-based platform designed to imitate human organ function, physiological flow, immune cells circulation, and inter-organ communication in vitro. The Fluidic Programmable Gravi-maze Array (FPGA) technology integrates multiple organ models, including gut, liver, kidney, brain, tumor, and vascular compartments, within a microfluidic architecture designed to reproduce physiologically relevant shear stresses and gravity-driven recirculating flow that facilitates inter-organ communication. Using computational fluid dynamics (CFD) simulations and impedance-based flow validation, we confirmed accurate shear control across organ compartments. Organ-specific and multiorgan models were constructed using 3D extracellular matrix hydrogels and assessed for metabolism, toxicity, and senescence. Liver-kidney co-cultures demonstrated metabolic interplay via differential albumin and urea production. In addition, the platform was evaluated for biologics testing using immune-oncology models incorporating tumor spheroids, endothelial barriers, and circulating immune cells. Antigen-specific T-cells, checkpoint inhibitors, bispecific antibody and antibody-drug conjugate (ADC) studies demonstrated the ability to measure on-target tumor killing, off-target toxicity, cytokine release, and bystander effects across interconnected tissue compartments under dynamic recirculating conditions. The system enabled longitudinal evaluation of immune-mediated cytotoxicity, tissue-selective responses, and cross-organ signaling not readily captured in conventional static assays. Overall, the OrganRX platform offers a physiologically relevant, scalable, and automation-compatible platform for preclinical drug evaluation, biologics safety assessment, and disease modeling. Its ability to capture complex, dynamic inter-organ effects position it as a powerful tool for advancing translational research, mechanistic toxicology, and precision medicine.

bioengineering↗