The Role of Perilipin 2 in Macrophages after Experimental Spinal Cord Injury

Perilipin 2 在实验性脊髓损伤后巨噬细胞中的作用

• 批准号: 10683728
• 负责人: Ethan Phares Glaser
• 金额: $ 5.27万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-09 至 2026-08-08
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10683728
• 关键词: Ablation; Acute; Alzheimer' s Disease; Arterial Fatty Streak; Atherosclerosis; Bone Marrow; Cells; Cholesterol; Cholesterol Homeostasis; Chronic; Cirrhosis; Custom; Cytosol; Data; Disease; Environment; Enzyme-Linked Immunosorbent Assay; Enzymes; Extramural Activities; Family; Fluorescent in Situ Hybridization; Foam Cells; Foundations; Functional disorder; Gene Expression; Genes; Genetic; Grant; Health Care Costs; Immunofluorescence Immunologic; In Vitro; Individual; Infiltration; Inflammation; Inflammatory; Injury; Interleukin-1 beta; Interleukin-6; Lesion; Leukocytes; Lipid-Laden Macrophage; Lipids; Locomotor Recovery; Macrophage; Mediating; Metabolism; Microscopy; Modeling; Morphology; Motor; Mus; Myelin; Myeloid Cells; Nervous System Trauma; Paralysed; Pathology; Pathway interactions; Phagocytosis; Phenotype; Physicians; Process; Production; Publishing; Quantitative Reverse Transcriptase PCR; Recovery; Recovery of Function; Research Support; Role; Scientist; Sensory; Site; Sorting; Spinal Cord; Spinal cord injury; Stains; Stimulus; System; Testing; Time; Tissues; Training; Transgenic Model; Up-Regulation; Visualization; Western Blotting; Work; collected works; cytokine; design; experimental study; histological stains; improved; in vitro Model; inflammatory milieu; injured; injury recovery; insight; lipid metabolism; molecular sequence database; monocyte; mouse model; nano-string; neuroinflammation; new therapeutic target; nonalcoholic steatohepatitis; novel; perilipin; prevent; repaired; response; responsible research conduct; single-cell RNA sequencing; skills; therapeutic target; uptake

PROJECT SUMMARY/ABSTRACT Spinal cord injury (SCI) leads to permanent motor and sensory loss that is exacerbated by persistent inflammation months after the injury. After SCI, monocyte-derived macrophages (MDMs) infiltrate the lesion to aid in cellular debris clearance. However, this response is emerging as a double- edged sword. Lingering debris inhibits repair and plasticity while clearance of the debris by infiltrating MDMs induces a proinflammatory and damaging phenotype. Clearance of cholesterol-rich myelin debris causes MDMs to resemble proinflammatory foam cells both phenotypically and morphologically. Foam cells are lipid- laden macrophages with numerous lipid droplets (LDs) that form when excessive cholesterol uptake overwhelms cholesterol metabolism. Foam cells also drive chronic inflammation in atherosclerosis and non- alcoholic steatohepatitis (NASH). Foam cells persist chronically in the injured cord and may contribute to the sustained proinflammatory lesion environment. However, their role in the chronic inflammatory state is poorly understood. Therefore, elucidating the effect of foam cell formation after SCI is integral to understanding the contribution of MDMs to injury pathology. Preliminary and published data identify Perilipin 2 (Plin2) as a key differentially upregulated gene in both infiltrating MDMs after SCI and foam cells in atherosclerosis. Plin2 is essential in regulating the formation and storage of cholesterol (i.e., lipid droplets (LDs)) in macrophages as lipid metabolites are either exported from these cells or stored in LDs. Plin2 coats LDs, sequesters them in the cytosol and prevents lipolytic enzymes from accessing their cargo. Reducing Plin2, may, therefore, reduce LD accumulation and mitigate foam cell formation after SCI. In models of atherosclerosis and NASH, Plin2 deficient mice produced fewer foam cells and were protected from atherosclerotic plaque formation and cirrhosis, respectively. To understand the effects of Plin2 upregulation in MDMs after SCI a novel targeted deletion of Plin2 using a LysMCre/lox system will be employed. Using this mouse model, this project will determine (1) the role of Plin2 in myelin-induced foam cell formation using an in vitro approach and (2) the effects of targeted Plin2 deletion in MDMs on foam cell formation, inflammation, and locomotor recovery after experimental. These experiments will test the hypothesis that cell-specific Plin2 ablation in MDMs suppresses foam cell formation after SCI, inhibits inflammation, and promotes functional recovery. If successful, this project will provide novel insight into how CNS macrophages metabolize myelin and may provide new therapeutic targets to improve SCI recovery by resolving chronic intraspinal inflammation. Through the completion of this training grant, the PI will receive ample training regarding the implementation of project management, responsible conduct of research, and the dissemination of findings. This training will provide the PI with the skillset necessary to carry out the specific aims and lay the foundation for the PI to become an independent physician-scientist with an extramurally supported research portfolio in the field of neurotrauma.

项目摘要/摘要脊髓损伤（SCI）导致永久电机和感官损失是 受伤几个月后的几个月后，由于持续的炎症而加剧。 SCI之后，单核细胞衍生的巨噬细胞 （MDMS）浸润病变以帮助细胞碎片清除。但是，这种反应正在成为双重的 边剑。挥之不去的碎屑抑制修复和可塑性，同时通过渗透MDMS清除碎屑 诱导促炎和破坏性表型。清除胆固醇富含髓鞘碎片的原因 MDM在表型和形态上类似于促炎泡沫细胞。泡沫细胞是脂质的 当胆固醇摄取过多时，载有许多脂质液滴（LD）的巨噬细胞 压倒性胆固醇代谢。泡沫细胞还驱动动脉粥样硬化和非 - 酒精性脂肪性肝炎（NASH）。泡沫细胞长期持续在受伤的绳索中，可能有助于 持续促炎病变环境。但是，它们在慢性炎症状态下的作用很差 理解。因此，阐明SCI后泡沫细胞形成的作用是理解 MDMS对损伤病理学的贡献。初步和已发表的数据将Perilipin 2（PLIN2）识别为关键 SCI和动脉粥样硬化中的SCI和泡沫细胞后浸润MDM的差异上调基因。 plin2是 调节胆固醇的形成和存储至关重要 脂质代谢物要么从这些细胞中导出，要么存储在LDS中。 plin2涂层LDS，将它们隔离在 细胞质并防止脂解酶进入其货物。因此，减少PLIN2，可能会减少LD SCI后积累并减轻泡沫细胞形成。在动脉粥样硬化和纳什的模型中，PLIN2 缺乏小鼠产生的泡沫细胞较少，并受到动脉粥样硬化斑块的形成和 肝硬化。要了解SCI后MDMS中PLIN2上调的影响 将使用lysmcre/lox系统删除PLIN2。使用此鼠标模型，此项目将 确定（1）使用体外方法和（2） MDM中有针对性的PLIN2缺失对泡沫细胞形成，炎症和运动恢复后的影响 实验。这些实验将检验以下假设：MDMS中的细胞特异性PLIN2消融抑制 SCI后的泡沫细胞形成，抑制炎症并促进功能恢复。如果成功，这个项目 将提供有关CNS巨噬细胞如何代谢髓磷脂的新颖洞察力，并可能提供新的治疗性 通过解决慢性脊柱内炎症来改善SCI恢复的靶标。通过完成 培训补助金，PI将获得有关实施项目管理的充分培训， 负责任的研究和发现的传播。该培训将为PI提供 执行特定目标所必需的技能，并为PI成为独立的基础 医师科学家在Neurotrauma领域具有外部支持的研究组合。

项目成果

Effects of Acute Ethanol Intoxication on Spinal Cord Injury Outcomes in Female Mice.

• DOI: 10.1089/neu.2023.0077
• 发表时间: 2023-12
• 期刊: JOURNAL OF NEUROTRAUMA
• 影响因子: 4.2
• 作者: Glaser, Ethan P.;Stewart, Andrew N.;Jagielo-Miller, Julia E.;Bailey, Caleb S.;Prendergast, Mark A.;Gensel, John C.
• 通讯作者: Gensel, John C.

Ethanol sustains phosphorylated tau protein in the cultured neonatal rat hippocampus: Implications for fetal alcohol spectrum disorders.

• DOI: 10.1016/j.alcohol.2022.07.007
• 发表时间: 2022-09
• 期刊: ALCOHOL
• 影响因子: 2.3
• 作者: Bailey, Caleb Seth;Jagielo-Miller, Julia Elaine;Keller, Peggy Sue;Glaser, Ethan Phares;Wilcox, Abigail Lowe;Prendergast, Mark Alan
• 通讯作者: Prendergast, Mark Alan