Mechanobiology of Viral Infections: Exploring Strain-Activated Calcium Channels in Lung Fibroblasts

Research funded in association with this project aims study how mechanical forces, such as changes in tissue stiffness and cyclic strain, impact viral infections. Using lung tissue as a model, it examines how physical changes from breathing or fibrosis affect virus-cell interactions. This research supports the national interest in advancing public health, as findings may guide safe activity levels during infections and suggest possible new treatments. The project also benefits society by offering research experiences for undergraduate and graduate students, with a focus on diversity. Outreach activities for K-12 students and the community will further support science education and inspire interest in research and health. The study will address two primary objectives to uncover mechanisms driving virus-host interactions under mechanical influences. Objective 1 investigates how matrix stiffness and cyclic strain affect viral infection rates and cellular responses in lung fibroblasts. Using custom-fabricated polydimethylsiloxane (PDMS) chambers with stiffness levels that mimic healthy and fibrotic lung tissue, CRL-4058 lung fibroblasts will be exposed to controlled mechanical strains that simulate breathing. Lung fibroblasts infected with vesicular stomatitis virus will be analyzed for viral infection rates (through RFU measurement, virus growth curve assays, Nucleocapsid (N) expression) and cellular behavior changes (through mRNA-seq, RT-qPCR, ELISA, Western Blot). Objective 2 examines the role of strain-activated calcium channels, such as TRP and Piezo channels in viral infection processes. By applying pharmacological inhibitors and monitoring calcium influx (Calbryte 520 AM) in response to mechanical strain, the study will first identify strain-activated calcium channels in CRL-4058 lung fibroblasts and then assess whether modulation of these identified channels affects viral replication and cell behavior. This investigation into the mechanobiology of viral infections seeks to provide new insights into the complex interplay between mechanical forces and viral pathogenesis in lung tissue. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.