Sex difference in conjunctival epithelial cell response and their profile of extracellular bioactive lipids duringallergic inflammation

过敏性炎症期间结膜上皮细胞反应的性别差异及其细胞外生物活性脂质的分布

• 批准号: 10749389
• 负责人: Changrim Lee
• 金额: $ 6.95万
• 依托单位: SCHEPENS EYE RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10749389
• 关键词: Affect; Allergens; Allergic; Allergic inflammation; Allergic rhinitis; American; Anabolism; Anterior eyeball segment structure; Asthma; Cell secretion; Cells; Conjunctival Epithelium; Cornea; Data; Dinoprostone; Disease; Eczema; Enzyme Activation; Enzymes; Eosinophilic Esophagitis; Epithelial Cells; Event; Extracellular Space; Eyelid structure; Female; Food Hypersensitivity; Goblet Cells; Health; Histamine; Histamine Release; Homeostasis; Human; Hypersensitivity; Incidence; Inflammation; Inflammation Mediators; Inflammatory; LOX gene; Leukotriene B4; Life; Link; Lipids; MAPK3 gene; Measures; Mediating; Mediator; Molecular; Mucins; PTGS2 gene; Phosphorylation; Phosphotransferases; Population; Prevalence; Publications; Reaction; Research; Research Proposals; Resolution; Role; Sclera; Severities; Sex Differences; Testing; Therapeutic; Time; Tissues; Vision; Visual; allergic response; comorbidity; comparison control; conjunctiva; extracellular; extracellular vesicles; human female; insight; lipid mediator; lipoxin A4; male; mast cell; novel; ocular surface; ocular surface disease; programmed cell death protein 1; response; sex; success

PROJECT SUMMARY Sex difference in the profile of extracellular bioactive lipid mediators, especially that of the specialized pro-resolving and proinflammatory lipid mediators (SPMs and PIMs, respectively), during inflammation is significantly correlated to sex differences in the cellular response, and prevalence and the severity of various inflammatory diseases, including ocular surface inflammation. As an ocular surface tissue, conjunctival epithelium lines the inside of the eyelids and covers the sclera, thus providing stability to the anterior segment of the eye. The conjunctival goblet cells (CGCs) secrete mucins to wet and protect the ocular surface, of which under- or over-secretion due to inflammation causes visual disturbance. Therefore, time-dependent resolution of conjunctival inflammation is critical for a transparent cornea and clear vision. Our exciting preliminary data show that CGCs secrete SPMs and PIMs as extracellular vesicle (EV) cargo to maintain homeostasis, and the amount of SPMs dramatically increases during a histamine-mediated allergic response. Interestingly, the increase was only observed in EVs of female CGCs but not males. EVs collected from males under the same condition showed a slight decrease in the amounts of SPMs compared to the control. To further investigate these novel findings, this research proposal aims to study the sex differences in the cellular response of human CGCs and the bioactive lipid mediators that CGCs secrete via EVs during histamine-mediated allergic inflammation. For Aim 1, we will identify sex differences in the cellular response by quantifying the magnitude of mucin secretion and change in intracellular Ca2+ concentration, and measuring the activation of intracellular enzymes involved in the allergic response and synthesis of bioactive lipid mediators. For Aim 2, we will determine the sex differences in the profile of extracellular bioactive lipid mediators during the allergic response by analyzing the type and amount of SPMs and PIMs loaded into EVs, and their time course of secretion. The success of this study will provide new insights into the bioactive lipid-mediated ocular surface protective mechanism generated by the conjunctiva and the basis for a tailored sex-dependent, lipid-based therapeutic approach in the management of vision-debilitating ocular surface diseases.

项目摘要 细胞外生物活性脂质介质谱的性别差异，特别是 促消退和促炎脂质介质（SPM和PIM，分别），在炎症过程中， 与细胞反应的性别差异显著相关，并且与各种 炎症性疾病，包括眼表炎症。 作为眼表组织，结膜上皮排列在眼睑内侧并覆盖巩膜， 从而为眼睛的前段提供稳定性。结膜杯状细胞分泌粘蛋白 湿润和保护眼表面，由于炎症引起的眼表面分泌不足或分泌过多会导致视力下降， 扰动因此，结膜炎症的时间依赖性消退对于透明角膜至关重要 清晰的视野。 我们令人兴奋的初步数据表明，CGCs分泌SPM和PIM作为细胞外囊泡（EV）货物 在组胺介导的过敏反应期间， 反应有趣的是，仅在女性CGCs的EV中观察到增加，而不是男性。收集的EV 在相同条件下从雄性中提取的样品与对照相比显示出SPM的量略有减少。 为了进一步研究这些新发现，本研究计划旨在研究 人CGC的细胞反应和CGC在EVS期间通过EV分泌的生物活性脂质介质 组胺介导的过敏性炎症。对于目标1，我们将通过以下方式确定细胞反应的性别差异： 定量粘蛋白分泌的大小和细胞内Ca2+浓度的变化，并测量细胞内Ca2+浓度， 参与过敏反应和生物活性脂质介质合成的细胞内酶的活化。 对于目标2，我们将确定在细胞外生物活性脂质介质的分布中的性别差异， 通过分析装载到EV中的SPM和PIM的类型和数量以及它们的时间过程， 分泌物。 这项研究的成功将为生物活性脂质介导的眼表提供新的见解 保护机制产生的结膜和基础上定制的性别依赖性，脂质为基础的 治疗方法在视力衰弱眼表疾病的管理。