Date of Submission

5-2026

Document Type

Thesis

Degree Name

Master of Science in Cellular and Molecular Biology

Department

Biology and Environmental Sciences

Advisor

Anna Kloc, Ph.D.

Committee Member

Saumya Ramanathan, Ph.D.

Committee Member

Alireza Senejani, Ph.D.

Keywords

Epstein-Barr Virus (EBV), Cardiovascular damage, Viral Lytic Pathways, EBV Encoded Viral Protein BHFR1, Transfection, Viral Sensor Protein NLRP3

MeSH

Herpesvirus 4, Human, Epstein-Barr Virus Infections, Cardiovascular Infections, Viral Proteins, Transfection, NLR Family, Pyrin Domain-Containing 3 Protein

LCSH

Epstein-Barr virus, Cardiovascular system, Viral proteins, Transfection

Abstract

Epstein-Barr Virus (EBV) is a ubiquitous gammaherpes virus characterized by its ability to establish viral persistence within host cells1-5. EBV is oncogenic and has the ability to shift between lytic and latent cycles1,10-13. EBV is implicated in a variety of disease states related to autoimmune conditions, mononucleosis, system lupus erythematosus (SLE), and multiple sclerosis (MS)1,2. Recent literature has sought to establish a mechanistic relationship between EBV and cardiovascular disease as EBV’s capacity to establish viral persistence within host cells seems to induce cardiovascular damage2, 5,. The underlying mechanism of this damage seems to be either directly through viral lytic pathways or indirectly through immune activation2. BHFR1 is a homolog of BCL2, a pro-survival protein in the human genome6. There is increasing evidence that BHFR1 drives chemoresistance via pro-apoptotic inhibition in cancers6; while promoting inflammasome activation to promote lytic activation6. Thus, this project hypothesizes that BHFR1 is sufficient to alter cell physiology by either driving unnatural cell proliferation (as seen in EBV related carcinomas) or promoting viral persistence and inducing indirect damage (in models of EBV induced cardiovascular disease). A cell culture model system of HEK293 cells was used to explore gene expression related to cell death and/or inflammation in BHFR1 transfected cells in comparison to non-BHFR1 bearing cells. Fluorescent imaging was taken at 24, 48, and 72 hours post transfection, and each timepoint was processed for nucleic acid extraction; cDNA synthesis of extracted mRNA; and qPCR analysis of target genes: NLRP3, Caspase 1, and Caspase 3. Gene expression for NLRP3 was downregulated in two biological replicates. One dataset showcased outlying NLRP3 expression at 24 and 72 hours post transfection. Caspase 3 expression was lowered across datasets; with higher expression at 24 hours. This coincides with analyzed BCL2 expression; which was expressed in all groups (the GFP only group was higher at 24 hours). Caspase 1 expression was temporal with decreased expression at 24 hours and an increase at 72 hours. These findings suggest that the protective effects commonly associated with BHFR1 might potentially be overcome, or rather that cells have some alternative way of rescuing the pyroptotic pathway in instances of decreased NLRP3 expression.

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