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.

项目概要/摘要该应用的长期目标是：(1) 确定每个单独的细胞类型如何衍生生理和病理条件下基底膜（BM）层粘连蛋白的变化， (2) 将这些层粘连蛋白改变与 AD 病理学相关联，以筛选独特的层粘连蛋白变化，对于早期 AD 诊断（痴呆或 Aβ/tau 病理发生前）和预后很有用该提案旨在产生一种创新的层粘连蛋白敲入小鼠品系，使其能够实现。以细胞特异性和 Cre 依赖性方式准确评估层粘连蛋白表达；确定每个单独细胞类型衍生的层粘连蛋白的临时和空间表达谱我们已经成功地生成了 Laminin-mCherry/eGFP 敲入。使用 CRISPR-Cas9 技术的小鼠品系，并在存在和存在的情况下进一步验证了这些小鼠在目标 1 中，我们将这些层粘连蛋白敲入小鼠与各种 Cre 杂交。使用这些 Lam-Rep 小鼠，产生一系列细胞特异性层粘连蛋白报告基因 (Lam-Rep) 小鼠。我们将确定大脑 BM 中层粘连蛋白的细胞来源，估计它们的相对丰度，并表征每个细胞类型衍生的层粘连蛋白的临时和空间表达谱在正常老化过程中，在目标 2 中，细胞特异性 Lam-Rep 小鼠将进入 5xFAD 背景。所得结果中每个细胞衍生的层粘连蛋白的临时和空间表达谱 Lam-Rep-5xFAD 小鼠及其非 AD Lam-Rep 同窝小鼠将按所述进行类似测定目标 1. 成功完成本研究将通过阐明以下内容来填补该领域的知识空白脑 BM 中层粘连蛋白的细胞来源并表征其临时/空间表达谱这些发现将在正常和 AD 条件下产生每种细胞类型衍生的层粘连蛋白。实现特定细胞类型来源的层粘连蛋白丢失与 AD 病理学之间的相关性研究；可以识别独特的层粘连蛋白变化，有助于早期 AD 诊断和预后预测。此外，该提案还将通过产生一个解决该领域进展的关键障碍创新的层粘连蛋白敲入小鼠系，允许在细胞特异性和 Cre- 中标记层粘连蛋白这种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）。和白细胞迁移）。这些研究将导致层粘连蛋白/BM 研究的突破。大大推动该领域向前发展。

项目成果

期刊论文数量（11）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

A dispensable role of oligodendrocyte-derived laminin-α5 in brain homeostasis and intracerebral hemorrhage

少突胶质细胞源性层粘连蛋白-α5 在脑稳态和脑出血中的不可或缺的作用

DOI：
10.1177/0271678x241228058
发表时间：
2024-01-19
期刊：
Journal of Cerebral Blood Flow & Metabolism
影响因子：
6.3
作者：
Minkyung Kang;A. Nirwane;Jingsong Ruan;Aravinthan Adithan;Marsilla Gray;Lingling Xu;Yao Yao
通讯作者：
Yao Yao

NR4A1 destabilizes TNF mRNA in microglia and modulates stroke outcomes.

NR4A1 破坏小胶质细胞中 TNF mRNA 的稳定性并调节中风结果。