Cell-specific changes of laminin expression in the CNS in Alzheimer’s disease

阿尔茨海默病中枢神经系统层粘连蛋白表达的细胞特异性变化

基本信息

批准号：
10283460
负责人：
Yao Yao
金额：
$ 41.11万
依托单位：
UNIVERSITY OF SOUTH FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-15 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10283460
关键词：
Address Affect Age Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease diagnosis Alzheimer&apos s disease pathology Amyloid beta-Protein Antibodies Area Astrocytes Basement membrane Blood - brain barrier anatomy Brain Brain region CRISPR/Cas technology Cells Communities Corpus striatum structure Correlation Studies Dementia Deposition Endothelial Cells Endothelium Event Extracellular Matrix Proteins Genetic Genetic Recombination Hematopoietic Hippocampus (Brain)Immunohistochemistry Individual Knock-in Mouse Knockout Mice Knowledge Label Laminin Leukocytes Maintenance Memory impairment Microglia Modeling Mus Neurodegenerative Disorders Neurons Oligodendroglia Pathogenesis Pathologic Pathology Pericytes Physiological Play Prognosis Protein Isoforms Reporter Research Research Personnel Role Series Severity of illness Source Techniques base blood-brain barrier disruption cell type conditional knockout disorder control in vivo imaging innovation normal aging success tau Proteins tool

项目摘要

Project Summary/Abstract The long-term objectives of this application are to: (1) determine how each individual cell type-derived laminin changes in the basement membrane (BM) under both physiological and pathological conditions, and (2) correlate these laminin alterations with AD pathology to screen for unique laminin changes that are useful in early AD diagnosis (before the onset of dementia or Aβ/tau pathology) and prognosis prediction. This proposal aims to generate an innovative laminin knock-in mouse line that enables accurate assessment of laminin expression in a cell-specific and Cre-dependent manner; and determine the temporary and spatial expression profile of each individual cell type-derived laminin in the BM in normal and AD brains. We have successfully generated the Laminin-mCherry/eGFP knock-in mouse line using CRISPR-Cas9 technique and further validated these mice in the presence and absence of Cre recombination. In Aim 1, we will cross these laminin knock-in mice with various Cre lines to generate a series of cell-specific laminin reporter (Lam-Rep) mice. Using these Lam-Rep mice, we will determine the cellular source of laminin in brain BM, estimate their relative abundance, and characterize the temporary and spatial expression profile of each individual cell type-derived laminin during normal aging. In Aim 2, cell-specific Lam-Rep mice will be crossed into the 5xFAD background. The temporary and spatial expression profile of each individual cell-derived laminin in the resulting Lam-Rep-5xFAD mice and their non-AD Lam-Rep littermates will be determined similarly as described in Aim 1. Successful completion of this study will fill the gap of knowledge in the field by elucidating the cellular source of laminin in brain BM and characterizing the temporary/spatial expression profile of each individual cell type-derived laminin under both normal and AD conditions. These findings will enable correlation studies between loss of specific cell type-derived laminin and AD pathology; and may identify unique laminin changes that are useful in early AD diagnosis and prognosis prediction. In addition, this proposal will also address a critical barrier to progress in the field by generated an innovative laminin knock-in mouse line, which allows labeling of laminin in a cell-specific and Cre- dependent manner. This genetic tool is also valuable to researchers in other fields (e.g. in vivo imaging and leukocyte transmigration). These studies will lead to a breakthrough in laminin/BM research and substantially move the field forward.

项目摘要/摘要该应用程序的长期目标是：（1）确定每个单独的单元类型衍生在物理和病理条件下，地下膜（BM）的层粘连蛋白变化，（2）将这些层粘连蛋白的改变与AD病理相关联，以筛选独特的层粘连蛋白变化在早期AD诊断（痴呆或Aβ/TAU病理学发作之前）和预后有用预言。该建议旨在产生创新的层粘连蛋白敲入鼠标线，以实现以细胞特异性和CRE依赖性方式准确评估层粘连蛋白表达；和确定每个细胞类型衍生的层粘连蛋白的临时和空间表达曲线正常和广告大脑中的BM。我们已经成功产生了层粘连蛋白 - 麦克利/EGFP敲击使用CRISPR-CAS9技术的鼠标线，并在存在的情况下进一步验证了这些小鼠没有CRE重组。在AIM 1中，我们将用各种CRE穿越这些层粘连蛋白敲入小鼠生成一系列细胞特异性层粘连蛋白报告基因（LAM-REP）小鼠的线。使用这些LAM-REP小鼠，我们将确定脑BM中层粘连蛋白的细胞来源，估计它们的相对丰度，并表征每个单个细胞类型衍生层粘连蛋白的临时和空间表达曲线在正常衰老期间。在AIM 2中，细胞特异性的LAM-REP小鼠将被跨入5xFAD背景。每个单独的细胞衍生的层粘连蛋白的临时和空间表达曲线 LAM-REP-5XFAD小鼠及其非AD LAM-REP文字将与所述相似地确定在AIM 1中。这项研究的成功完成将通过阐明该领域的知识差距脑BM中层粘连蛋白的细胞来源，并表征在正常和AD条件下，每个单独的细胞类型衍生的层粘连蛋白。这些发现会在特定细胞类型衍生的层粘连蛋白和AD病理学的丧失之间启用相关研究；和可以识别独特的层粘连蛋白变化，这些变化可用于早期AD诊断和预后预测。在此外，该建议还将通过生成的创新的层粘连蛋白敲入小鼠系，它允许在细胞特异性和Cre-中标记层粘连蛋白依赖方式。该遗传工具对于其他领域的研究人员也很有价值（例如体内成像和白细胞传播）。这些研究将导致层粘连蛋白/BM研究的突破实质上将田地向前推动。