[R21] Screening for small-molecule inhibitors of respiratory syncytial viral infection in infant bronchial epithelial barrier
Ente: National Institute of Allergy and Infectious Diseases
Scadenza: 2028-08-31
Importo max: $233,250
Paese: US
Descrizione
PROJECT SUMMARY
Respiratory syncytial virus (RSV) primarily infects ciliated cells in the human bronchial epithelium. RSV infection
can cause severe disease in infants and children younger than 2 years of age. So far, RSV vaccine
development has been unsuccessful and treatment of severe RSV disease in young children remains
supportive care. In contrast, RSV-infected adults have only mild symptoms. In a recent study, we found that the
human bronchial epithelial barrier plays a critical role in determining severity of RSV infection with age.
Preliminary single-nuclear RNA sequencing of RSV-infected, neonatal and adult ciliated cells has identified
multiple signaling pathways in response to RSV that differ between the two age groups. These preliminary
findings lead to the central hypothesis that small-molecule inhibitors of the signaling pathways regulated by
age in infant ciliated cells in response to RSV infection can be identified by high-throughput screening as
potential therapeutics. To this end, we have established a high-throughput screening platform using RSVGFP
pre-infected neonatal bronchial epithelial cells generated from basal stem cells (BSCs) in a 96-transwell format.
The proposal leverages this platform to screen a L2100 stem cell signaling pathway compound library that
features 1243 compounds with bioactivities overlapping age-related signaling responses to RSV in neonatal
ciliated cells. Our screening is facilitated by a robust method of BSC derivation from tracheal aspirate, which
circumvents technical difficulties in accessing infant lung tissues required for conventional BSC isolation. To
complement human bronchial epithelial cells as an in vitro model, precision-cut lung slices (PCLSs) prepared
from infant donor lungs will be employed as an ex vivo model. We have designed a stage-screening strategy
including primary, counter, and secondary screenings followed by mechanistic investigation of top-ranked
compounds. This project is led by Dr. Ai (PI, lung development and airway hyperactive disease in children
(asthma and RSV infection)) in consultation with Dr. Krishnan (airway physiology and high-throughput drug
screening). Dr. Ai and Dr. Krishnan are long-term collaborators and together have established several methods
and established the imaging platform employed for the proposed library screening. The project is expected to
identify candidate compounds for future therapeutic testing using pre-clinical animal models of infant RSV
disease. The findings of epithelium-derived mechanisms of RSV infection may be translatable to other
respiratory viruses that elicit similar age-related infection and thus inform the fundamental biology of age in
susceptibility and severity of human respiratory diseases.
Istituzione: WASHINGTON UNIVERSITY
PI: Xingbin Ai
Progetto: 1R21AI196505-01A1
Settori: National Institute of Allergy and Infectious Diseases
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