Investigating the Impact of Soat1 Knock-Out on Visceral Adipose Tissue Regulatory T-Cell Accumulation in TCR Transgenic Mice. Restricted; Files Only

Abstract

Visceral adipose tissue (VAT) regulatory T cells (Tregs) are a subset of FOXP3⁺, CD4⁺ T cells localized in the visceral adipose tissue. These cells help maintain tissue homeostasis within the VAT and regulate organismal metabolism. Our research attempts to shed light on how modulating membrane cholesterol levels would influence the accumulation of VAT Tregs, which play important roles in limiting obesity-associated inflammation and metabolic syndrome. We attempt to answer this question by knocking out the Soat1 gene, which catalyzes the formation of cholesterol-esters from free cholesterol, in TCR transgenic Tregs and performing a competitive transfer assay into recipient mice to measure the Soat1 KO Treg accumulation in the visceral adipose tissue. We hypothesize that knocking out Soat1 will increase Treg accumulation in the visceral adipose tissue due to higher free cholesterol levels in the cell membrane, which will result in more T-cell receptor clustering and signaling. However, our results showed no significant accumulation of Soat1 KO donor Tregs within the VAT and suggests that Soat1 function of sequestering free cholesterol into storage as cholesterol esters may not be substantial enough by itself to alter the lipid raft composition of these cells. 

Table of Contents

Introduction: 1

Materials and Methods: 4

Table 1. Soat1-targeting sgRNAs’ sequences: 4

Results: 7

Figure 1. Flow cytometry gating strategy: 7

Figure 2. Soat1 sgRNA percent knock-out efficiencies: 9

Figure 3. Percentage of donor-derived Tregs in recipient mice: 10

Figure 4. Expression levels of Soat1 and Soat2 in Tregs of vTreg53 TCR-tg mice: 11

Discussion: 11

References: 14

About this Honors Thesis

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School	Emory College
Department	Biology
Degree	B.S.
Submission	Honors Thesis
Language	English
Research Field	Biology, Cell
Keyword	Immunology Regulatory T cells
Committee Chair / Thesis Advisor	Li, Chaoran, Emory University
Committee Members	Catano, Laura, Emory University Jacob, Nitya, Emory University

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