Endothelial-Pericyte Crosstalk in Diabetic Stroke

糖尿病中风的内皮-周细胞串扰

• 批准号: 10400506
• 负责人: Nabil J Alkayed
• 金额: $ 20.48万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10400506
• 关键词: Acute; Anti-Inflammatory Agents; Blood Vessels; Blood capillaries; Blood flow; Brain; CSPG4 gene; Cells; Chronic Phase; Complement; Cytometry; Dementia; Development; Diabetes Mellitus; Disease; DsRed; Endothelium; Female; GPR39 gene; Gene Expression Profiling; Heterogeneity; Impaired cognition; Inflammatory; Injury; Ischemia; Label; Lead; Morphology; Mus; National Institute of Neurological Disorders and Stroke; Neuronal Dysfunction; Outcome; Parents; Pericytes; Phase; Phenotype; Play; Process; Proliferating; Recovery; Reporting; Research; Role; Signal Transduction; Stroke; Testing; Tissue-Specific Gene Expression; Work; brain cell; diabetic; insight; male; neuroinflammation; non-diabetic; parent project; post stroke; post stroke cognitive impairment; post stroke dementia; preservation; prevent; repaired; single-cell RNA sequencing; stroke recovery; stroke risk; therapy development; transcriptome sequencing

Abstract Work proposed in the current supplement complements research proposed in the parent NINDS R01 NS108501- 01 “Endothelial-Pericyte Crosstalk in Diabetic Stroke”. Studies in the parent R01 were focused on the acute vascular changes that are common after diabetic stroke (i.e., microvascular no-reflow). The current supplement will investigate the long-term cognitive impairment that follows stroke and that is also more common after diabetic stroke (post-stroke cognitive impairment and dementia, PSCID). The parent project specifically investigated the role of endothelial-pericyte signaling (endothelial-derived epoxyeicosatrienoates (EETs) acting on GPR39 on peri-capillary pericytes) in microvascular no-reflow after diabetic stroke. We tested the hypothesis that EETs/CPR39 crosstalk preserves capillary blood flow in brain after ischemia, and that diabetes reduces microvascular endothelial EETs, leading to pericyte contraction and injury and subsequent capillary occlusion (no-reflow). During the course of the study, we made the observation that in the delayed phase after stroke, pericytes detach from capillaries, proliferate and migrate away from capillaries. We have also observed that these pericytes undergo morphological transformation that is consistent with pericyte activation. Activated pericytes have been reported to play a pro-inflammatory as well anti-inflammatory role under different disease conditions. However, the role of pericyte activation during the chronic phase of recovery from stroke and in post- stroke cognitive impairment under diabetic conditions is not clear. We will test the hypothesis that in normal brain, pericyte activation promotes recovery and functional repair after stroke. Under diabetic conditions, activated pericytes switch to a pro-inflammatory phenotype that contributes to neuroinflammation and neuronal dysfunction underlying PSCID. We propose to conduct single-cell RNAseq (scRNAseq) and single-cell mass cytometry (scCyTOF) of pericytes isolated from male and female, diabetic and no-diabetic NG2-DsRed mice, which label pericytes in red, at 1 day (when pericytes are still attached to capillaries) and 7 days after stroke (when pericytes are activated and detached from capillaries). Single-cell RNAseq will provide new information on pericyte heterogeneity after stroke under diabetic and non-diabetic conditions (on a spectrum ranging from quiescent, capillary attached pericytes to activated, migrating and proliferating pericytes). Furthermore, differential gene expression analysis of pericyte subpopulations from diabetic and non-diabetic brains will provide insight into the role of pericyte heterogeneity in recovery from stroke and post-stroke dementia. We will confirm RNAseq results and further phenotype pericyte subpopulations using scCyTOF.

抽象的 在当前补充填充完成的研究中提出的工作在父母Ninds R01 NS108501-中提出的工作 01“糖尿病中风中的内皮 - 肽串扰”。在父r01中的研究集中于急性 糖尿病性中风后常见的血管变化（即微血管无流量）。当前补充 将研究中风后的长期认知障碍，这在糖尿病后也更为常见 中风（冲程后认知障碍和痴呆，PSCID）。家长项目专门调查了 内皮 - 周期性信号传导的作用（内皮衍生的环氧树阳离子（EET）作用于GPR39上的Eets（EET） 糖尿病性中风后微血管无流量的细胞周期周期）。我们检验了以下假设 EET/CPR39串扰可保留缺血后大脑中毛细血管血流，糖尿病减少 微血管内皮EET，导致周细胞收缩和受伤以及随后的毛细血管阻塞 （无回流）。在研究过程中，我们观察到，在中风后的延迟阶段， 周细胞脱离毛细血管，扩散并远离毛细血管。我们还观察到 这些周细胞经历与周细胞激活一致的形态转化。活性 据报道，周细胞在不同疾病下发挥促炎和抗炎作用 状况。但是，周细胞激活在中风恢复的慢性阶段和后 糖尿病状况下的中风认知障碍尚不清楚。我们将测试以下假设 大脑，周细胞激活可促进中风后的恢复和功能修复。在糖尿病状况下， 活化的周细胞切换到有助于神经炎症和神经元的促炎表型 PSCID的功能障碍。我们建议进行单细胞RNASEQ（SCRNASEQ）和单细胞质量 从男性和女性，糖尿病和无糖尿病NG2-DSRED小鼠分离的周细胞的细胞仪（SCCYTOF）， 在1天（毛细管附着周细胞时）和中风后7天，哪个标记为红色 （激活周细胞并与毛细血管脱离时）。单细胞RNASEQ将提供新信息 在糖尿病和非糖尿病状况下中风后的周细胞异质性（在光谱上 静止的，毛细管附着于活化，迁移和增殖的周细胞）。此外， 糖尿病和非糖尿病大脑的周细胞亚群的差异基因表达分析将提供 深入了解周细胞异质性在中风和中风后痴呆中恢复中的作用。我们将确认 RNASEQ使用SCCYTOF结果和进一步的表型细节亚群。

项目成果

Role of GPR39 in Neurovascular Homeostasis and Disease.

• DOI: 10.3390/ijms22158200
• 发表时间: 2021-07-30
• 期刊: International journal of molecular sciences
• 影响因子: 5.6
• 作者: Xu Y;Barnes AP;Alkayed NJ
• 通讯作者: Alkayed NJ

Apolipoprotein E4 mediates insulin resistance-associated cerebrovascular dysfunction and the post-prandial response.

• DOI: 10.1177/0271678x17746186
• 发表时间: 2019-05
• 期刊: Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism
• 作者: Johnson LA;Torres ER;Weber Boutros S;Patel E;Akinyeke T;Alkayed NJ;Raber J
• 通讯作者: Raber J

GPR39 Deficiency Impairs Memory and Alters Oxylipins and Inflammatory Cytokines Without Affecting Cerebral Blood Flow in a High-Fat Diet Mouse Model of Cognitive Impairment.

• DOI: 10.3389/fncel.2022.893030
• 发表时间: 2022
• 期刊: Frontiers in cellular neuroscience
• 影响因子: 5.3

Vascular Biology.

• DOI: 10.1161/strokeaha.121.033556
• 发表时间: 2021-07
• 期刊: Stroke
• 影响因子: 8.3
• 作者: Alkayed NJ;Cipolla MJ
• 通讯作者: Cipolla MJ

Age-dependent cognitive impairment, hydrocephalus and leukocyte infiltration in transgenic mice with endothelial expression of human EPHX2.

• DOI: 10.1038/s41514-022-00090-1
• 发表时间: 2022-07-05
• 期刊: NPJ AGING
• 作者: Davis, Catherine M;Zhang, Wenri H;Bah, Thierno M;Roese, Natalie E;Allen, Elyse M;Leung, Philberta;Boutros, Sydney J;Marzulla, Tessa;Patel, Esha;Nie, Xiao;Alkayed, Farah N;Huang, Justin H;Jensen, Michael A;Raber, Jacob;Pike, Martin M;Alkayed, Nabil J
• 通讯作者: Alkayed, Nabil J