Modeling of Huntington's Disease Astrocytes from Non-human Primate Neural Progenitor Cells Pubblico

Abstract

Huntington's disease (HD) is an autosomal dominant disease characterized by an overexpansion of CAG repeats in the mutant huntingtin gene (mHTT). Recently, increasing evidence has shown that astrocytes, a type of glial cells responsible for the mediation of neural and glial functions and the structural integrity of the blood brain barriers, can accumulate mutant huntingtin protein and compromise their neural supportive functions. Animal models have provided a useful approach to study the pathological mechanism of HD. However, deficiencies still exist in these models, causing incomplete or erroneous recapitulation of the HD phenotype. To further minimize the difference between animal models and HD patients, this study employed neural progenitor cells (NPCs) reprogrammed from rhesus monkeys' induced pluripotent cells (iPSCs) as a base-stone to derive astrocytes in vitro. To create a HD monkey model, monkeys were transfected with exon1 with 72 CAG repeats in mHTT. In comparison to iPSCs, NPCs exhibited a greater capacity to generate a homogenous neural population and mirror the HD characteristics. Results of this study implied that HD astrocytes derived from monkey NPCs could recapitulate the down-regulation of PGC1-alpha, GLT-1, and SOD2 found in HD patients, as well as the up-regulation of glutamate receptors and the existence of intranuclear inclusions, which indicated the presence of poly-glutamine aggregates as an essential HD phenotype. A dramatic alteration in astrocyte morphology was also perceived. Nonetheless, this HD astrocyte model revealed phenotypes unobserved in HD patients in terms of increased BDNF expression, which might be attributed to calcium-induced dysregulation of gene transcription.

Table of Contents

Chapter 1: Background

1.1 Overview of Huntington's Disease (HD)...1

1.2 Genetics and Pathological Mechanisms of Huntington's Disease (HD)...3

1.3 Astrocytes in Neurodegenerative Disease...7

1.4 Animal Models to Recapitulate HD Symptoms and Pathogenic Mechanism...12

Chapter 2: HD Astrocyte Modeling

2.1 Introduction...15

2.1 Hypotheses...18

2.2 Material and Methods

2.2.1 Preparation of A NPC Monolayer in Vitro...19

2.2.2 Astrocyte Differentiation from NPCs...19

2.2.3 Immunocytochemistry...20

Table 1 A-B...21

2.2.4 Real-time PCR to Measure Astrocytic Gene Expression...22

Table 2...22

2.2.5 Oxidative Challenge with H2O2...23

2.3 Results

2.3.1 Apoptotic and Morphological Differences in HD and wild type NPC derived astrocytes...24

Figure 1...25

2.3.2 Expression of Astrocytic Markers...26

Figure 2-Figure 6...28-29

2.3.3 Real-time PCR on Classic Markers for Astrocyte Function...30

Figure 7...31

2.3.4 Oxidative Challenge with H2O2...32

Figure 8...33

Figure 9-Figure 10...34

2.4 Discussion...35

2.5 Conclusion...41

Reference...42

About this Honors Thesis

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School	Emory College
Department	Biology
Degree	BS
Submission	Honors Thesis
Language	English
Research Field	Biology, Neuroscience
Parola chiave	Huntington's Disease
Committee Chair / Thesis Advisor	Chan, Anthony, Emory University
Committee Members	Eisen, Arri, Emory University Cafferty, Patrick William, Emory University

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