VCOM Louisiana Research Day Program

Biomedical Research

Kristina McLeod-van Amstel, OMS-II; Dr. Rebecca Morrow, PhD Edward Via College of Osteopathic Medicine-Louisiana 08 CHARACTERIZING THE ROLE OF AMYLOID BETA IN PULMONARY ENDOTHELIUM AS PART OF THE INFLAMMATORY IMMUNE RESPONSE.

Context: Pseudomonas aeruginosa has been identified as a key infectious organism in the development of pneumonia and sepsis. During infection, the immune system causes pulmonary endothelium to release inflammatory cytokines along with amyloid proteins. certain strains of P. aeruginosa can disrupt the host cell defense and disable the pulmonary endothelium’s autophagy ability, allowing for pathogenic tau and amyloid proteins to accumulate without recycling. Of the amyloid proteins involved, levels of amyloid beta have been shown to increase in concert with the infection as well as decrease upon remission. While tau is known to be cytotoxic to cells and contributory to the pathology of the infection, the role of amyloid beta as it contributes to the inflammatory immune response has yet to be characterized within the pulmonary system. As such, this study aims to understand its role as it contributes to innate host defense within pulmonary health using rat pulmonary vascular endothelial cells and by targeting its precursor protein APP. Objective and/or Hypothesis: The goal of this study is to determine the role of amyloid beta in pulmonary endothelial cells by deleting its precursor protein APP and determining if there is any change in behavior between control and knockout cells in both the normal and

physiological challenge states.

Methods: Pulmonary vascular endothelial cells came from the rat cell line MVR1D. Two clones of cells with CRISPR/Cas9 deleted APP will be compared to the wild type MVR1D and a gRNA cell line that will act as a CRISPR control. The expression of IL-6, TNF, CCL5, and IL-1 β genes in each cell line will be determined using RT-PCR. Cell lines will be cultured in standard medium and assessed for growth through an MTT assay. Later endeavors include infecting the cells with P. aeruginosa and assessing their inflammatory reaction via a cellular cytokine assay. Results/Anticipated Results: The study is currently in progress and results are not yet complete. It is anticipated that the APPKO cell lines will grow at similar rates as the wild type; however, they may lack the robust inflammatory reaction that the wild type would. Conclusion(s): Conclusions not yet reached due to ongoing data analysis.

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