Is this project an undergraduate, graduate, or faculty project?
Undergraduate
Project Type
individual
Campus
Daytona Beach
Authors' Class Standing
Riley Dienna, Senior
Lead Presenter's Name
Riley Dienna
Lead Presenter's College
DB College of Arts and Sciences
Faculty Mentor Name
Dr. Amber Paul
Abstract
During spaceflight, the immune system of astronauts becomes impaired due to microgravity, resulting in increased susceptibility to infections caused by pathogens such as Candida albicans. One of the most common clinical treatments for aggressive candidiasis is amphotericin B, a polyene antimycotic that targeted ergosterol in the fungal cell membrane and compromised the stability of the membrane. However, due to the chemical similarities between ergosterol and cholesterol, this treatment also affects the somatic cells of patients it is administered to. A novel treatment is therefore required to combat the aggressive fungal infection while minimizing damage to the host’s cells. This project aimed to create a novel treatment for C. albicans by utilizing an AAV1 plasmid vector to knock in endo-alpha-mannosidase, an enzyme from Bacteroides thetaiotaomicron—a member of the gut microbiome that regulated the growth of C. albicans in healthy individuals—into macrophage-like cells differentiated from the U937 cell line. The cells were exposed to simulated microgravity and then differentiated with 100 nM of phorbol 12-myristate 13-acetate (PMA), followed by transfection with the donor vector. Evaluation of cell surface markers, reactive oxygen species, and phagocytic activity was completed via flow cytometry. The cells exposed to simulated microgravity and transfected with the donor vector were hypothesized to exhibit increased expression of cell surface markers associated with phagocytosis as well as enhanced phagocytic activity. Data analysis has revealed that there were not statistically significant changes in cell surface expression or phagocytic activity in the transfected cells. Future directions of this project aim to examine the possibility of utilizing bacteriophage technology in the gut microbiome to engineer an immune response to prevent the dysregulation of the microbiome that allows for commensal organisms, such as C. albicans, to become dangerous opportunistic pathogens.
Did this research project receive funding support (Spark, SURF, Research Abroad, Student Internal Grants, Collaborative, Climbing, or Ignite Grants) from the Office of Undergraduate Research?
No
Gene Therapy Strategies to Enhance Antifungal Immunity in Microgravity
During spaceflight, the immune system of astronauts becomes impaired due to microgravity, resulting in increased susceptibility to infections caused by pathogens such as Candida albicans. One of the most common clinical treatments for aggressive candidiasis is amphotericin B, a polyene antimycotic that targeted ergosterol in the fungal cell membrane and compromised the stability of the membrane. However, due to the chemical similarities between ergosterol and cholesterol, this treatment also affects the somatic cells of patients it is administered to. A novel treatment is therefore required to combat the aggressive fungal infection while minimizing damage to the host’s cells. This project aimed to create a novel treatment for C. albicans by utilizing an AAV1 plasmid vector to knock in endo-alpha-mannosidase, an enzyme from Bacteroides thetaiotaomicron—a member of the gut microbiome that regulated the growth of C. albicans in healthy individuals—into macrophage-like cells differentiated from the U937 cell line. The cells were exposed to simulated microgravity and then differentiated with 100 nM of phorbol 12-myristate 13-acetate (PMA), followed by transfection with the donor vector. Evaluation of cell surface markers, reactive oxygen species, and phagocytic activity was completed via flow cytometry. The cells exposed to simulated microgravity and transfected with the donor vector were hypothesized to exhibit increased expression of cell surface markers associated with phagocytosis as well as enhanced phagocytic activity. Data analysis has revealed that there were not statistically significant changes in cell surface expression or phagocytic activity in the transfected cells. Future directions of this project aim to examine the possibility of utilizing bacteriophage technology in the gut microbiome to engineer an immune response to prevent the dysregulation of the microbiome that allows for commensal organisms, such as C. albicans, to become dangerous opportunistic pathogens.