PhAIge Therapy Center: AI-Guided Tailspike Engineering and Molecular Typing for Rapid Phage Design and Testing Against ESKAPE Pathogens

PROJECT SUMMARY/ABSTRACT The Gladstone PhAIge Therapy Center seeks to address the critical threat of multidrug-resistant ESKAPE pathogens by developing next-generation preclinical assays and computational tools. Our overarching goal is to establish a robust, quantitative, and predictive preclinical framework that accelerates the pipeline from phage discovery to clinical application, with a primary focus on Klebsiella pneumoniae. Specifically, Project 1 develops high-throughput, sequencing-based assays to measure rate constants for each component of the phage infection cycle (recognition, entry, replication, packaging, and lysis). By capturing granular, quantitative data, this work will enable the rational selection and engineering of phages that exhibit superior efficacy. Project 2 employs multiplexed assays to characterize K. pneumoniae strains rapidly by their capsule types and susceptibility profiles. Integrating capsule-typing with systematic phage compatibility testing will allow for the creation of optimized, off-the-shelf phage cocktails tailored to individual clinical isolates. The Center's strategy includes two specialized Cores to ensure translational relevance and efficiency. The Advanced Computational Core combines bioinformatics pipelines, AI-driven modeling, and machine learning to predict phage-host interactions and guide phage selection. The Organoid and Human Cell Culture Core incorporates 3D human tissue models and primary cell cultures into early screening, capturing host-pathogen dynamics that are overlooked by standard in vitro assays. An Administrative Core fosters collaboration, manages the Development and Research Program (DRP) to support innovative pilot projects, and liaises closely with clinical and industry advisors. By uniting modern molecular methods, computational analytics, and human- relevant models, the Gladstone PhAIge Therapy Center will advance systematic, clinically ready phage therapies to combat antibiotic resistance.