Engineering Fetal Cell Exosomes to contain HMGB1: Its trafficking and role as an inflammatory activator in uterine cells

工程化胎儿细胞外泌体以含有 HMGB1：其运输及其在子宫细胞中作为炎症激活剂的作用

• 批准号: 9883717
• 负责人: RAMKUMAR MENON
• 金额: $ 14.55万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-03-01 至 2022-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9883717
• 关键词: Amniotic Fluid; Animal Model; Animals; Biochemical; Biogenesis; Biological; Birth; Cell Aging; Cell Culture Techniques; Cell membrane; Cells; Communication; Complex; Cytoplasm; Data; Data Reporting; Decidua; Distant; Dose; Encapsulated; Endocrine; Endosomes; Engineering; Epithelial Cells; Feedback; Fetal Membranes; Fetal Tissues; Fetus; Future; Generations; Genes; HMGB1 Protein; HMGB1 gene; Image; In Vitro; Induced Labor; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injections; Injury; Intervention; Label; Labor Onset; Laboratories; Leukocytes; Light; Liquid substance; Maternal-Fetal Exchange; Mediating; Membrane; Methods; Modeling; Molecular; Mothers; Mus; Myometrial; NF-kappa B; Nuclear; Nuclear Protein; Optics; Oxidative Stress; Paracrine Communication; Pathway interactions; Pattern; Phosphorylation; Physiological; Plasma; Pregnancy; Premature Birth; Premature Labor; Process; Production; Property; Proteins; Publishing; Reporting; Reproduction; Risk; Role; Sampling; Series; Side; Signal Transduction; Site; Sterility; Stimulus; Technology; Testing; Tissues; Uterus; Vesicle; amnion; base; chemokine; cytokine; engineered exosomes; exosome; extracellular; extracellular vesicles; fetal; fetus cell; maternal serum; monocyte; mouse model; myometrium; new technology; novel; p38 Mitogen Activated Protein Kinase; particle; pregnant; prenatal stress; protein protein interaction; response; senescence; therapeutic protein; tissue stress; trafficking; vesicular release

ABSTRACT Feto-maternal paracrine signaling, an inflammatory process, is one of the key initiators of parturition. Recently we reported that senescent fetal tissues at term, specifically fetal membrane cells, generate damage- associated molecular pattern markers (DAMPs) with proinflammatory properties, locally and at distant sites. DAMPs, along with other inflammatory cytokines and chemokines, enhance the inflammatory load to transition quiescent uterine tissues to an active state for parturition. DAMPs can reach distant sites through exosomes (30–150-nm extracellular vesicles) released by senescent fetal cells and cause inflammatory changes by increasing cytokine and chemokine productions in the recipient cells. One of the well-studied DAMPs in term and preterm parturition is high-mobility group box (HMGB)1 protein. HMGB1 is a non-histone nuclear protein, released to the cytoplasm because of nuclear injury due to cellular senescence or other insults. Senescence of fetal amnion epithelial cells (AEC) leads to packaging of HMGB1 in exosomes. HMGB1 concentration is higher in senescent-cell-derived exosomes, which led us to hypothesize that AEC exosomes containing HMGB1 can either systemically or by traversing through tissues reach maternal uterine cells (decidua and myometrium), cause functional changes by enhancing their inflammatory load (NF-kB activation and increased production of cytokines), and contribute to the parturition process. Current studies have not been able to elucidate the functional capabilities of exosome-encapsulated HMGB1 because senescent AEC exosomes traffic contents other than HMGB1. To get a precise functional property of HMGB1 in exosomes, we plan to engineer exosomes to contain a novel protein via the Exosomes for Protein Loading Via Optically Reversible Protein- Protein Interactions (EXPLORs) method. This method is developed to overcome the limitations of conventional exosome-based protein delivery. Using these HMGB1-encapsulated exosomes, we will test HMGB1's functional role in vitro in decidual and myometrial cell cultures and in animal models of pregnancy. Our aims will be Specific Aim #1: to engineer AEC-derived exosomes to specifically contain HMGB1 by integrating a reversible protein-protein interaction module controlled by blue light with the endogenous process of exosome biogenesis. Specific Aim #2: to test the functional role of HMGB1-rich AEC-derived exosomes in in vitro and pregnant animal models. We expect to generate exosomes containing specific cargo and test its functional role in promoting inflammation and parturition. Successful generation of exosomes loaded with specific cargo will introduce a new technology to load exosomes with functionally viable proteins. Future studies will test the usefulness of exosomes with therapeutic proteins that can be used for interventional purposes in adverse pregnancies.

摘要 胎儿-母体旁分泌信号是一种炎症过程，是分娩的关键启动子之一。最近 我们报道了足月时衰老的胎儿组织，特别是胎膜细胞，会产生损伤， 与局部和远端部位的促炎特性相关的分子模式标志物（DAMP）。 DAMPs，沿着其他炎性细胞因子和趋化因子，增强炎症负荷， 从静止的子宫组织到分娩的活跃状态。DAMP可以通过外泌体到达遥远的位点 （30 - 150-nm的细胞外囊泡）由衰老的胎儿细胞释放，并通过 增加受体细胞中细胞因子和趋化因子的产生。长期以来研究得很好的DAMP之一 和早产是高迁移率族蛋白（HMGB）1蛋白。HMGB1是一种非组蛋白核蛋白， 由于细胞衰老或其他损伤引起的核损伤而释放到细胞质中。衰老 胎儿羊膜上皮细胞（AEC）导致HMGB1包装在外泌体中。HMGB1浓度较高 在衰老细胞来源的外泌体中，这使我们假设含有HMGB1的AEC外泌体可以 或者全身地或者通过穿过组织到达母体子宫细胞（蜕膜和子宫肌层）， 通过增强其炎症负荷（NF-kB活化和增加的 细胞因子），并有助于分娩过程。目前的研究还不能阐明 外泌体包裹的HMGB1的功能能力，因为衰老的AEC外泌体运输内容物 除了HMGB1。为了获得HMGB1在外泌体中的精确功能特性，我们计划工程化 外泌体通过用于通过光学可逆蛋白质进行蛋白质装载的外泌体含有新蛋白质- 蛋白质相互作用（EXPLORs）方法。这种方法是为了克服传统方法的局限性而开发的。 基于外泌体的蛋白质递送。使用这些HMGB1包裹的外泌体，我们将测试HMGB1的 在体外蜕膜和子宫肌层细胞培养物和妊娠动物模型中的功能作用。我们的目标 将 具体目标#1：通过整合可逆的HMGB1基因，工程化AEC衍生的外泌体以特异性地含有HMGB1。 蓝光调控的蛋白质-蛋白质相互作用模块与exosome的内源过程 生物起源 具体目标#2：测试富含HMGB1的AEC衍生的外泌体在体外和妊娠中的功能作用。 动物模型 我们期望产生含有特定货物的外泌体，并测试其在促进细胞增殖中的功能作用。 炎症和分娩。装载有特定货物的外泌体的成功产生将引入一种新的免疫系统。 一种新技术，将功能可行的蛋白质装载到外泌体上。未来的研究将测试 具有治疗性蛋白质的外泌体可用于不良妊娠的干预目的。

项目成果

Extracellular vesicles from maternal uterine cells exposed to risk factors cause fetal inflammatory response.

• DOI: 10.1186/s12964-021-00782-3
• 发表时间: 2021-10-07
• 期刊: Cell communication and signaling : CCS
• 作者: Shepherd MC;Radnaa E;Tantengco OA;Kechichian T;Urrabaz-Garza R;Kammala AK;Sheller-Miller S;Menon R
• 通讯作者: Menon R