Kaili Coenen

Document Type

Honors Thesis

Degree Name

Bachelor of Science

Date of Award

Spring 5-1-2025

Abstract

X-linked chronic granulomatous disease (XCGD) is a debilitating immunodeficiency disorder primarily affecting men. The disease results from mutations in the CYBB gene on the X chromosome, which encodes the GP91^PHOX protein responsible for electron transfer from NADPH to molecular oxygen. Despite the identification of the CYBB mutation, the pathological mechanism underlying the reduction in reactive oxygen species (ROS) generation in XCGD remains unclear. We hypothesized that comparative gene expression analysis, combined with pathway analysis, could identify genes and signaling pathways contributing to reduced reactive oxygen species (ROS) levels in XCGD. In this study, we analyzed gene expression data from four healthy subjects and eight XCGD patients. Five genes—IGKC, ID2, RAB11FIP3, APOC1, and FCER1A—were identified as significantly altered (≥10-fold change, p < .05), suggesting their potential involvement in XCGD. Notably, ID2 expression exhibited a 4.21-fold reduction, implying a role in ROS production. To further investigate this, we compared ID2 and RAB11FIP3 expressions in ROSlow and ROS-high cells. In qRT-PCR analysis, ID2 mRNA levels were reduced 1.8-fold in lowROS MCF-7/G1P3 cells compared to vector controls. Additionally, treatment with N-Acetylcysteine (NAC), a known ROS scavenger, significantly increased ID2 mRNA expression in MCF-7/Vector cells (p = .0031). RAB11FIP3 levels were upregulated in both high-ROS and low-ROS cell lines. Taken together, our findings suggest that in XCGD, ROS regulates the expression of ID2 and RAB11FIP3, which in turn play a role in ROS generation. Restoring ID2 and RAB11FIP3 expressions may enhance ROS production and T-cell proliferation, offering potential therapeutic targets for managing XCGD.

Advisor

Venu Cheriyath

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Keywords

Reactive oxygen species (ROS), ID2, RAB11FIP3, APOC1, X-linked Chronic Granulomatous Disease (XCGD), CYBB, NADPH, gp91phox, Gene analysis, cDNA Synthesis, Reverse transcriptase, Primer Preparation, Quantitative Reverse Transcriptase-Polymerase Chain Reaction (qRT-PCR) Analysis, Nacetylcysteine (NAC), PBS, Complete media, trypsin

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