Regulatory Network Modeling of Cellular Transitions in Aging Epithelia and Capillaries

During epithelial wound healing, senescence makes short-term use of damaged cells by blocking their division while they boost survival, proliferation, and migration in their neighbors. Meanwhile, partial Epithelial to Mesenchymal Transition (EMT) creates migrating and proliferating cells that close the wound. Aging is known to disrupt wound healing, but designing effective interventions is limited by a lack of quantitative computational tools that reproduce and predict the mechanisms by which aging disrupts tissue healing. This requires multi- scale models in which individual cell behaviors are driven by detailed molecular regulatory networks. The objective of this proposal is to build and experimentally validate single-cell network models that reproduce the environment-dependent commitment of epithelial cells to senescence vs. hybrid E/M vs. mesenchymal state as a function of age. The central hypothesis is that aging micro-environments disrupt the coordination between hybrid E/M and senescence in healing epithelia by promoting full EMT, blocking its reversal, and speeding senescence in proliferating cells. This ultimately delays wound healing in epithelial sheets (Aim 1) and disrupts spouting angiogenesis (Aim 2). The hypothesis is based on results from preliminary models that reproduce partial vs. full EMT determined by biomechanical cues and synthesize mechanisms of senescence in response to DNA damage. The rationale is that unified regulatory models of the full repertoire of cell behaviors observed in aging wounds will transform our qualitative understanding of aging mechanisms into a tool to predict new interventions. The main hypothesis will be tested in three Specific Aims designed to build and validate models that reproduce 1) mutually exclusive commitment to senescence vs. the mesenchymal state in age-dependent epithelial healing, and 2) angiogenic pattern formation, EMT, and senescence in healthy vs. aging capillaries. 3) A third aim integrates model validation into a Course-Based Undergraduate Research Experience (CURE), where students learn Boolean modeling and contribute to a database of model /published experiment comparisons. Our approach centers dynamically modular Boolean modeling, innovative due to its focus on regulatory switches within larger networks that trigger discrete cell-state changes. This resulted in software that translates molecular events into changing combinations of cell behaviors and can automate validation against molecular and phenotypic data. The proposed work is significant for undergraduate research at Wooster, as the funding will help scaffolds computational biology research into our curriculum: introduction via CURE labs, in-depth training via sophomore research, and publishable research in senior Independent Study. The project strengthens research collaboration and professional opportunities for College of Wooster students across the life sciences. The project is significant for biomedical research because it is expected to generate in silico cells for tissue models, which can aid therapeutic design in COPD, lung cancer, and diseases with pathological angiogenesis. As tissues pay for healing by aging, the ultimate goal is to speed healing while lowering its cost. Project Number: 1R15AG083621-01A1 | Fiscal Year: 2026 | NIH Institute/Center: National Institute on Aging (NIA) | Principal Investigator: Erzsebet Regan | Institution: COLLEGE OF WOOSTER, WOOSTER, OH | Award Amount: $482,620 | Activity Code: R15 | Study Section: Special Emphasis Panel[ZRG1 BBBT-M (84)] View on NIH RePORTER: https://reporter.nih.gov/project-details/11291738