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Variant-to-Function Mapping of Late Onset Alzheimer's Disease GWAS Loci in Human Microglial Models Implicates RTFDC1 at the CASS4 Locus Elizabeth Anne Burton
- Format:
- Book
- Thesis/Dissertation
- Author/Creator:
- Burton, Elizabeth Anne, author.
- Language:
- English
- Subjects (All):
- Genetics.
- Cellular biology.
- Neurosciences.
- Molecular biology.
- Immunology.
- 0369.
- 0379.
- 0982.
- 0317.
- 0307.
- Local Subjects:
- Genetics.
- Cellular biology.
- Neurosciences.
- Molecular biology.
- Immunology.
- 0369.
- 0379.
- 0982.
- 0317.
- 0307.
- Physical Description:
- 1 electronic resource (166 pages)
- Contained In:
- Dissertations Abstracts International 86-07B
- Place of Publication:
- Ann Arbor : ProQuest Dissertations and Theses, 2024
- Language Note:
- English
- Summary:
- Late-onset Alzheimer's disease (LOAD) research has principally focused on neurons over the years due to their known role in the production of amyloid beta plaques and neurofibrillary tangles. In contrast, recent genomic studies of LOAD have implicated microglia as culprits of the prolonged inflammation exacerbating the neurodegeneration observed in patient brains. Indeed, recent LOAD genome-wide association studies (GWAS) have reported multiple loci near genes related to microglial function, including TREM2, ABI3, and CR1. However, GWAS alone cannot pinpoint underlying causal variants or effector genes at such loci, as most signals reside in non-coding regions of the genome and could presumably confer their influence frequently via long-range regulatory interactions. The goal of this thesis was to identify novel LOAD GWAS-associated variants and their associated regulatory elements, and characterize the effects of one variant-regulatory element pair in a microglial cell model. To this end, we carried out a combination of ATAC-seq and high-resolution promoter-focused Capture-C in two human microglial cell models (iPSC-derived microglia and HMC3) in order to physically map interactions between LOAD GWAS-implicated candidate causal variants and their corresponding putative effector genes. Notably, we observed consistent evidence that rs6024870 at the GWAS CASS4 locus contacted the promoter of nearby gene, RTFDC1. We subsequently observed a directionally consistent decrease in RTFDC1 expression with the protective minor A allele of rs6024870 via both luciferase assays in HMC3 cells and expression studies in primary human microglia. Through CRISPR-Cas9-mediated deletion of the putative regulatory region harboring rs6024870 in HMC3 cells, we observed increased pro-inflammatory cytokine secretion and decreased DNA double strand break repair related, at least in part, to RTFDC1 expression levels. This variant-to-function approach therefore reveals that the rs6024870-harboring regulatory element at the LOAD 'CASS4' GWAS locus influences both microglial inflammatory capacity and DNA damage resolution, along with cumulative evidence implicating RTFDC1 as a novel candidate effector gene
- Notes:
- Source: Dissertations Abstracts International, Volume: 86-07, Section: B.
- Includes supplementary digital materials
- Advisors: Grant, Struan F. A.; Brown, Christopher D. Committee members: Wang, Li-San; Aquizu-Lopez, Naiara; Lee, Edward; Ritchie, Marylyn
- Ph.D. University of Pennsylvania 2024
- Local Notes:
- School code: 0175
- ISBN:
- 9798302184054
- Access Restriction:
- Restricted for use by site license
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