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Principal Investigator: Lily Charpentier
Organization: DARTMOUTH COLLEGE
Fiscal Year: 2024
Award: $48,974
Funding agency: National Heart Lung and Blood Institute
ABSTRACT
Cystic fibrosis (CF) is characterized by the formation of thick mucus in the lung leading to chronic
infection of various pathogens that are prone to developing antibiotic tolerance. Despite effective
modulator therapy (HEMT) that improves lung function and many clinical outcomes, HEMT does
not eliminate chronic, antibiotic-tolerant lung infections, the major cause of morbidity and mortality
in CF. Although most people with CF (pwCF) have multispecies lung infections, almost all
published studies focus on single-species infection models. Thus, polymicrobial-host interactions
in CF are not completely understood, especially under physiologically relevant conditions such as
anoxia typical of mucus plugs.
The overarching goal of our work is to use in vitro models to gain insight into how polymicrobial
infections develop in pwCF. Bacterial pathogens have been shown to secrete membrane vesicles
(bEVs) that diffuse through the CF mucus to deliver virulence factors such as DNA, RNA, and
proteins to their targets. As recent studies have shown that the first exposure to bacterial products
alters DNA methylation and gene silencing that affect subsequent exposures, our hypothesis is
that pwCF develop chronic infections in part due to epigenetic changes caused by preexposure
to CF pathogens that reduce the HBEC immune response to infection over time. This application
aims to characterize the effects of an in vitro model of CF polymicrobial infection on human
bronchial epithelial cells (HBEC) with preexposure to bEVs secreted by the common CF pathogen
Staphylococcus aureus. The polymicrobial culture contains four prevalent and abundant CF
pathogens: Pseudomonas aeruginosa, S. aureus, Streptococcus sanguinis, and Prevotella
melanogenica. Using transcriptomic and proteomic analysis, cytokine analysis, ATAC-Seq, and
DNA methylation analysis, this study aims to elucidate the host HBEC response to treatment with
bEVs. This study will contribute to our understanding of host-pathogen interactions in the CF lung
and potentially identify novel therapeutic targets during infection.
This project will provide the applicant with a broad range of both bioinformatic and lab techniques
that will provide a strong foundation for her long-term goal of becoming an academic researcher.
Terms: <ATAC sequencing><ATAC-seq><ATACseq><Affect><Anoxia><Antibiotic Agents><Antibiotic Drugs><Antibiotic Resistance><Antibiotics><Assay for Transposase-Accessible Chromatin using sequencing><Bacteria><Bacteroides melaninogenicus><Bio-Informatics><Bioinformatics><CF lung disease><CF mucus><CFTR><CFTR Protein><Cell secretion><Cellular Secretion><Chronic><Clinical><Clinical Data><Collaborations><Communication><Cystic Fibrosis><Cystic Fibrosis Transmembrane Conductance Regulator><DNA><DNA Methylation><DNA analysis><Data><Deoxyribonucleic Acid><Development><Diffuse><Environment><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Epithelial Cells><Exposure to><Foundations><Gene Expression><Gene Inactivation><Gene Silencing><Gene Transcription><Generalized Growth><Genetic Transcription><Goals><Growth><Human><Immune><Immune response><Immunes><Immunological response><Infection><Investigators><Life><Lung><Lung Respiratory System><Lung infections><Measures><Membrane><Miscellaneous Antibiotic><Modeling><Modern Man><Morbidity><Morbidity - disease rate><Mucins><Mucociliary Clearance><Mucociliary Transport><Mucous body substance><Mucoviscidosis><Mucus><Mucus Glycoprotein><Non-Polyadenylated RNA><O element><O2 element><Outcome><Oxygen><P aeruginosa><P melaninogenicus><P. aeruginosa><P. melaninogenicus><Pathogenicity Factors><Persons><Phenotype><Physiologic><Physiological><Prevotella><Prevotella melaninogenica><Prevotella melaninogenicus><Proteins><Proteome><Proteomics><Pseudomonas aeruginosa><Pseudomonas pyocyanea><Publishing><Pulmonary Cystic Fibrosis><RNA><RNA Expression><RNA Gene Products><Research Design><Research Personnel><Researchers><Resistance to antibiotics><Resistant to antibiotics><Ribonucleic Acid><S aureus><S sanguinis><S sanguis><S. aureus><S. aureus infection><S. sanguinis><S. sanguis><Secondary to><Short interfering RNA><Small Interfering RNA><Sputum><Staph aureus><Staph aureus infection><Staphylococcus aureus><Staphylococcus aureus infection><Streptococcus sanguinis><Streptococcus sanguis><Study Type><Techniques><Testing><Thick><Thickness><Time><Tissue Growth><Training><Transcription><Vesicle><Virulence><Virulence Factors><Work><analyze DNA><antibiotic drug resistance><antibiotic resistant><antibiotic tolerance><assay for transposase accessible chromatin followed by sequencing><assay for transposase accessible chromatin seq><assay for transposase accessible chromatin sequencing><assay for transposase-accessible chromatin with sequencing><bacteria pathogen><bacterial pathogen><bronchial epithelium><chronic infection><cystic fibrosis lung><cystic fibrosis lung disease><cystic fibrosis mucus><cystic fibrosis transmembrane regulator><cytokine><deliver short interfering RNA><deliver siRNA><deliver small interfering RNA><delivery system for siRNA><delivery system for small interfering RNA><delivery vectors for siRNA><deprivation><developmental><epigenetically><experience><exposed human population><extracellular vesicles><host response><human exposure><immune system response><immunoresponse><improved><in vitro Model><infected with S. aureus><infected with Staph aureus><infected with Staphylococcus aureus><insight><lung function><membrane structure><methylation pattern><mortality><mucoid><mucous><new drug target><new drug treatments><new druggable target><new drugs><new pharmacological therapeutic><new pharmacotherapy target><new therapeutic target><new therapeutics><new therapy><new therapy target><next generation therapeutics><novel><novel drug target><novel drug treatments><novel druggable target><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel pharmacotherapy target><novel therapeutic target><novel therapeutics><novel therapy><novel therapy target><ontogeny><pathogen><pathogenic bacteria><persistent infection><pulmonary><pulmonary function><pulmonary infections><response><response to therapy><response to treatment><secondary infection><short interfering RNA delivery><siRNA><siRNA delivery><skills><small interfering RNA delivery><study design><therapeutic response><therapy response><tolerance to antibiotics><tolerate antibiotics><transcriptional silencing><transcriptomics><treatment response><treatment responsiveness>