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.

摘要 当前补充中提出的工作补充了母NINDS R 01 NS 108501中提出的研究- 01“糖尿病卒中中的内皮细胞-周细胞串扰”。在母体R 01中的研究集中于急性 糖尿病中风后常见的血管变化（即，微血管无复流）。当前增补 将研究中风后的长期认知障碍，这在糖尿病患者中也更常见。 中风（中风后认知障碍和痴呆，PSCID）。父项目专门调查了 内皮-周细胞信号传导（内皮衍生的环氧二十碳三烯酸（ESTs）作用于GPR 39， 毛细血管周细胞）在糖尿病卒中后微血管无复流中的作用。我们测试的假设 缺血后EAE/CPR 39串扰保护脑中的毛细血管血流量，并且糖尿病降低了 微血管内皮细胞增生，导致周细胞收缩和损伤以及随后的毛细血管闭塞 （无复流）。在研究过程中，我们观察到在中风后的延迟期， 周细胞从毛细血管分离、增殖并从毛细血管迁移。我们还注意到， 这些周细胞经历与周细胞活化一致的形态学转化。激活 据报道，周细胞在不同的疾病中发挥促炎和抗炎作用 条件然而，周细胞活化在中风后恢复的慢性期和中风后恢复期的作用是不确定的。 糖尿病条件下的中风认知损害尚不清楚。我们将检验在正常情况下 脑，周细胞活化促进中风后的恢复和功能修复。在糖尿病条件下， 活化的周细胞转变为促炎表型， PSCID潜在的功能障碍。我们建议进行单细胞RNAseq（scRNAseq）和单细胞质量 从雄性和雌性、糖尿病和非糖尿病NG 2-DsRed小鼠分离的周细胞的细胞计数（scCyTOF）， 在中风后第1天（周细胞仍然附着在毛细血管上）和第7天， (when周细胞被激活并从毛细血管脱离）。单细胞RNAseq将提供新的信息 在糖尿病和非糖尿病条件下中风后周细胞异质性的影响（范围从 静止的、毛细血管附着的周细胞到活化的、迁移的和增殖的周细胞）。此外，委员会认为， 糖尿病和非糖尿病脑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恢复中的作用。我们将确认 使用scCyTOF的RNAseq结果和进一步表型周细胞亚群。

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

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.

GPR39缺乏症会损害记忆力并改变催产素和炎症细胞因子，而不会影响高脂饮食小鼠认知障碍模型中的脑血流。

• 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

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

载脂蛋白E4介导与胰岛素抵抗相关的脑血管功能障碍和餐后反应。

• 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

Plasma Oxylipins: A Potential Risk Assessment Tool in Atherosclerotic Coronary Artery Disease.

• DOI: 10.3389/fcvm.2021.645786
• 发表时间: 2021
• 期刊: Frontiers in cardiovascular medicine
• 影响因子: 3.6
• 作者: Le DE;García-Jaramillo M;Bobe G;Alcazar Magana A;Vaswani A;Minnier J;Jump DB;Rinkevich D;Alkayed NJ;Maier CS;Kaul S
• 通讯作者: Kaul S