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dc.contributor.advisorMichael J. Leeen_US
dc.contributor.authorHoneywell, Megan E
dc.date.accessioned2024-01-04T19:07:36Z
dc.date.available2024-01-04T19:07:36Z
dc.date.issued2023-12-18
dc.identifier.doi10.13028/d4ph-k012
dc.identifier.urihttp://hdl.handle.net/20.500.14038/52909
dc.description.abstractA common approach to understanding how drugs induce their therapeutic effect is identifying the genetic determinants of drug sensitivity. This can be achieved following systematic loss- or gain-of-function genetic perturbations with CRISPR/Cas9. Because these “chemo-genetic profiles” are generally performed in a pooled format, inference of gene function is subject to several confounding influences, including variation in growth rates between clones or variation in the degree of coordination between growth and death. To overcome these issues, we developed an analysis method called MEDUSA (Method for Evaluating Death Using a Simulation-assisted Approach). MEDUSA uses time-resolved measurements and model driven constraints to reveal the combination of growth and death rates that generated the drug-treated clonal abundance. We find that MEDUSA is uniquely effective at identifying death regulatory genes, and we apply MEDUSA to determine how DNA damage-induced lethality varies in the presence and absence of p53. We find that loss of p53 switches the mechanism of DNA damage-induced death from apoptosis to a non-apoptotic form of death called MPT-driven necrosis. We find that activation of MPT by DNA damage requires high respiration, and that cell death can be exacerbated by modulating NAD+ in p53-deficient cells. These findings demonstrate the accuracy and utility of MEDUSA, both for determining the genetic dependencies of lethality and for revealing opportunities to promote the lethality of chemotherapies in a cancer specific manner.en_US
dc.language.isoen_USen_US
dc.publisherUMass Chan Medical Schoolen_US
dc.rightsCopyright © 2023 Megan E. Honeywellen_US
dc.rights.uriAll Rights Reserveden_US
dc.subjectCell deathen_US
dc.subjectChemo-genetic profilingen_US
dc.subjectCRISPR screeningen_US
dc.subjectMEDUSAen_US
dc.subjectMEDUSA(GM)en_US
dc.subjectp53en_US
dc.subjectMPT-dependent necrosisen_US
dc.subjectApoptosisen_US
dc.subjectNon-apoptotic deathen_US
dc.subjectELP complexen_US
dc.subjectEGFR inhibitorsen_US
dc.subjectMCL1en_US
dc.titleExperimental and Computational Methods for Identifying Death-Regulatory Genes from Chemo-Genetic Profilesen_US
dc.typeDoctoral Dissertationen_US
atmire.contributor.authoremailmegan.honeywell@umassmed.eduen_US
dc.contributor.departmentSystems Biologyen_US
dc.description.thesisprogramInterdisciplinaryen_US
dc.identifier.orcid0000-0002-4894-8672en_US


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