Modeling Alzheimer's Disease in Hispanic Latino populations using human cortical organoids

使用人类皮质类器官模拟西班牙裔拉丁裔人群的阿尔茨海默病

• 批准号: 10680168
• 负责人: Karina K Meyer-Acosta
• 金额: $ 3.89万
• 依托单位: UNIVERSITY OF TEXAS SAN ANTONIO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10680168
• 关键词: 3-Dimensional; Abeta synthesis; Address; Alleles; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease related dementia; Alzheimer' s disease risk; Biological Assay; Biological Models; Biology; Brain; Cell Death; Cells; Central America; Cerebrum; Cessation of life; Characteristics; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Disease; Disease model; Dorsal; Enzyme-Linked Immunosorbent Assay; Ethnic Origin; Ethnic Population; Fellowship; Gene Expression; Genes; Genetic; Genetic Risk; Genetic Transcription; Genetic Variation; Genetic study; Genome; Genotype; Geography; Growth; Heritability; Hispanic; Hispanic ancestry; Human; Immunohistochemistry; Impaired cognition; Impairment; In Vitro; Individual; Latino Population; Learning; Measures; Mediating; Methods; Mexican; Mexican Americans; Molecular; Neuroglia; Neurons; Not Hispanic or Latino; Organoids; Pathologic; Pathology; Pathway interactions; Patients; Pattern; Phenotype; Physiological; Population; Population Heterogeneity; Proliferating; Proteins; Risk; Role; Scientist; South American; Support Groups; Testing; Training; Variant; Western Blotting; apolipoprotein E-3; apolipoprotein E-4; cell type; differential expression; disease phenotype; disorder risk; ethnic diversity; excitatory neuron; experimental study; genetic manipulation; genetic risk factor; genome wide association study; health disparity; high risk; induced pluripotent stem cell; induced pluripotent stem cell technology; inhibitory neuron; insight; multi-ethnic; new therapeutic target; novel; protective effect; protein expression; risk variant; skills; stem; stem cell technology; tau-1; therapeutic target; transcriptome sequencing

Project Summary/Abstract Hispanic Latinos (HLs) have the highest risk of developing Alzheimer’s disease (AD) and related dementias, the highest proportion of any ethnic group in the US. In genome-wide association studies (GWAS) conducted mostly in non-Hispanic white (NHW) individuals, Apolipoprotein E4 (APOE4) was established as the leading AD genetic risk factor, increasing risk 3-12 fold compared to the non-risk allele, APOE3. In HLs however, there is discordant evidence about the association of APOE4 with AD which is at least partially attributed to the genetic diversity of HLs stemming from widespread geographic origins. GWAS in large HL populations identified ancestry native to Central America, the primary ancestry of Hispanic Mexican Americans (HMA), have little to no AD association with APOE4. Studies further suggest that inheritance of APOE4 allele from native ancestor may not increase AD risk to the same extent as that observed in NHW. It is postulated that genetic variation, possibly regulatory, either local to APOE allele, or across the genome, modifies APOE4 biology. Recently, genetic variation local to APOE was shown to dominantly influence gene transcription and APOE expression independent of APOE genotype. Gaining insight on the molecular basis by which HMA background modifies APOE4 biology would identify protective mechanisms against AD. My central hypothesis is that APOE4 will be associated with AD-related phenotypes in cortical organoids with NHW ancestry, but not with HMA ancestry, or to a lesser extent. Since AD develops decades before cognitive decline and ethnic variation is human-specific, I will test my hypothesis in 3D cortical organoids. I will generate excitatory neuron-enriched human cortical organoids (hCO), and inhibitory neuron-enriched human subpallial organoids (hSO) to explore neuron subtype-specific differences in AD-related phenotypes. To investigate cellular phenotypes mediated by APOE4 with respect to genetic background, I will 1) characterize the effect of APOE4 on AD-related cellular phenotypes within NHW background, and 2) determine the effect of HMA background on APOE4 mediated phenotypes and gene transcription. My preliminary data suggests that the APOE4 allele reduces growth and upregulates APOE expression in both hCO and hSO compared to organoids carrying the non-risk allele, APOE3. Aim 1 will explore the cellular basis contributing to size (differentiation, maturation, and cell death) and characterize AD-related pathology by immunostaining and protein expression assays. To control for patient background and assess the specific effects of APOE4 genotype and genetic background, I will gene-edit APOE3 to APOE4 in iPSCs with NHW and HMA backgrounds. In Aim 2, I will determine the phenotypic and transcriptional changes associated with HMA background on APOE4- mediated AD-related phenotypes. Under this fellowship, I will expand my molecular toolkit and in vitro disease modeling and hone my skills as a molecular neuroscientist. I will also strengthen my analytical skills and learn new methods to probe gene expression by training under my sponsor and a supporting group of scientists.

项目摘要/摘要 拉美裔拉美裔(HLS)患阿尔茨海默病(AD)和相关痴呆的风险最高， 这是美国所有种族中比例最高的。在主要进行的全基因组关联研究中 在非西班牙裔白人(Nhw)中，载脂蛋白E4(APOE4)被确定为AD的主要基因 风险因素，与非风险等位基因APOE3相比，风险增加3-12倍。然而，在HLS中，存在着不和谐 载脂蛋白4与阿尔茨海默病关联的证据至少部分归因于 HLS起源于广泛的地理来源。在大量的HL人群中发现了原产于 中美洲是西班牙裔墨西哥裔美国人(HMA)的主要祖先，几乎没有AD的联系 使用APOE4。研究进一步表明，APOE4等位基因遗传自原生祖先可能不会增加AD 风险程度与在NHW观察到的程度相同。据推测，基因变异，可能是调节性的，或者 APOE等位基因的局部或整个基因组改变了APOE4的生物学特性。最近，APOE的局部遗传变异 对基因转录和APOE表达的影响不受APOE基因型的影响。 深入了解HMA背景修饰APOE4生物学的分子基础将识别 对AD的保护机制。我的中心假设是，APOE4将与AD相关 皮质细胞器的表型具有nhw血统，但不具有hma血统，或者程度较小。自公元后 在认知衰退和种族差异是人类特有的之前几十年就会发生，我将在3D中测试我的假设 皮质类器官。我会产生兴奋性神经元丰富的人类皮质有机化合物(HCO)，并抑制 神经元富集人大脑皮层下有机体(HSO)对AD相关神经元亚型特异性差异的探讨 表型。为了研究APOE4介导的细胞表型与遗传背景的关系，我将 1)在nhw背景下表征APOE4对AD相关细胞表型的影响；2) 确定HMA背景对APOE4介导的表型和基因转录的影响。我的初选 数据表明，APOE4等位基因降低了HCO和HSO的生长速度，上调了APOE的表达 与携带非危险等位基因APOE3的有机化合物相比。目标1将探索细胞基础对 大小(分化、成熟和细胞死亡)，并通过免疫染色和 蛋白质表达分析。对患者背景进行对照并评估APOE4基因的特异性效应 和遗传背景，我将在nhw和hma背景的ipscs中编辑APOE3到APOE4的基因。在AIM 2，我将确定与APOE4上HMA背景相关的表型和转录变化- 介导的AD相关表型。在这个奖学金的帮助下，我将扩展我的分子工具箱和体外疾病 作为一名分子神经学家，我的建模和技能得到了磨练。我也会加强我的分析能力，学习 在我的赞助人和一群科学家的支持下，通过培训来探索基因表达的新方法。