Improving Brain Recovery Through Glycoengineering

通过糖工程改善大脑恢复

基本信息

批准号：
10666616
负责人：
Xiaofeng Jia
金额：
$ 48.23万
依托单位：
UNIVERSITY OF MARYLAND BALTIMORE
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10666616
关键词：
Adhesions Affinity Animal Model Animals Apoptosis Behavior assessment Behavioral Biochemical Biological Biological Process Biology Brain Brain Injuries Cadherins Carbohydrates Cell Adhesion Cell Communication Cell surface Cells Cellular biology Central Nervous System Diseases Chemical Structure Chemicals Chemistry Complex Development Dose Electrophysiology (science)Engineering Event Extracellular Matrix Foundations Generations Glycoconjugates Glycoengineering Gold Healthcare Heart Arrest Human Implant In Vitro Intercept Ischemia Kinetics Lead Malignant Neoplasms Mass Spectrum Analysis Mediator Metabolic Metabolism Methodology Modeling Monosaccharides N-acetylmannosamine Nature Nerve Regeneration Neurites Neuronal Differentiation Neurons Neurophysiology - biologic function Organism Pathway interactions Pharmaceutical Preparations Physiological Polysaccharides Preclinical Testing Proteins Rattus Recovery Recovery of Function Reporting Rodent Model Safety Sialic Acids Signal Transduction Spatial Distribution Stimulus Structure-Activity Relationship Sulfhydryl Compounds Surface Synaptic plasticity Techniques Testing Therapeutic Time Tissues Translations Transplantation WNT Signaling Pathway Wnt proteins Work analog blastomere structure cell motility clinical application clinical translation conventional therapy cost effectiveness covalent bond design drug candidate embryo cell extracellular functional group functional improvement glycoproteomics healing immune cell infiltrate improved in vivo injured injury recovery innovation intercellular communication migration nerve stem cell neural neuroregulation novel receptor response scaffold stem cell biology stem cell differentiation stem cell fate stem cell therapy technology platform therapeutic candidate translational approach

项目摘要

Project Summary This project is based on recent advances in metabolic glycoengineering (MGE), a technology platform where non-natural monosaccharides intercept the biosynthetic pathways for cell surface-displayed glycans. As a result, chemical functionalities not naturally found in carbohydrates are installed in the glycalyx, which can alter cell adhesion, receptor activity, and downstream events (e.g., apoptosis, differentiation, and motility). In previous work, we developed the N-acetylmannosamine (ManNAc) analog “Ac5ManNTGc” to install thiol groups into sialic acids in human embryonic cells and found that – when the cells were grown on a “high affinity” surface (e.g., gold, which forms coordinate covalent bonds with thiols) – Wnt signaling was upregulated in the absence of extracellular Wnt proteins and neuronal differentiation was induced. In vivo translation of this approach, however, was hindered by the requirement for a non-degradable gold scaffold. We recently overcame this impediment by designing new ManNAc analogs with thiols presented on longer linkers, which extends this functional group further away from the core monosccharide and increases analog potency. Critically, the new analogs provide pro-neurogenic activity in the absence of a scaffold thereby simplifying in vivo translation. This project will explore analog mechanism in hNSCs in Specific Aim 1; this aim will define how the chemical structure, kinetics, and dose of thiol-modified ManNAc analogs (along with chemically inert size-matched controls) modulate cellular glycans in Aim 1a; evaluate changes to cell adhesion and motility in Aim 1b, and evaluate the differentiation of human neural stem cells (hNSCs) in Aim 1c. Next, in Specific Aim 2, we will apply the optimized analog-treatment conditions to improve neural regeneration in a rat cardiac arrest (CA) model of brain injury by transplanting MGE-modified into injured animals. We will compare hNSCs treated with our new thiol-modified analogs with appropriate controls on functional recovery after CA by evaluating survival, adhesion, distribution, and migration of transplanted hNSCs in rat brain. In Specific Aim 3, we will evaluate biochemical (Wnt signaling and cadherin involvement) and cellular (tissue infiltrating immune cells) level mechanisms we propose contribute to the healing effects of MGE in brain injury recovery (in Aim 3a). Finally, in Aim 3b we will characterize cell-wide “glycosites” by mass spectrometry and use glycobioinformatics analyses to identify unknown biochemical mediators of MGE. Specifically, we anticipate identifying new mediators of the beneficial effects of MGE in the implanted hNSCs as well as trans-acting host proteins. We hypothesize that thio-analogs modulate hNSC fate through a complex combination of receptor- specific effects on cell signaling and adhesion providing a pleiotropic suite of healing effects that cannot be achieved through conventional therapies. Accordingly, our innovative approach opens a new avenue to improve stem cell therapy with our new thiol-based MGE technique.

项目概要该项目基于代谢糖工程（MGE）的最新进展，这是一个技术平台，其中非天然单糖拦截细胞表面展示聚糖的生物合成途径。结果，碳水化合物中不存在的化学功能被安装在糖萼中，这可以改变细胞粘附、受体活性和下游事件（例如细胞凋亡、分化和运动）。在之前的工作中，我们开发了N-乙酰甘露糖胺（ManNAc）类似物“Ac5ManNTGc”来安装硫醇在人类胚胎细胞中分组为唾液酸，并发现当细胞以“高亲和力”生长时表面（例如金，与硫醇形成配位共价键）——Wnt 信号传导在细胞外 Wnt 蛋白的缺失和神经元分化被诱导。然而，这种方法受到了对不可降解金支架的需求的阻碍。通过设计新的 ManNAc 类似物，其硫醇存在于较长的连接体上，克服了这一障碍，将该功能组扩展至远离核心单糖的位置并增加模拟效力。重要的是，新的类似物在没有支架的情况下提供了促神经发生活性，从而简化了该项目将在具体目标 1 中探索 hNSC 中的模拟机制；硫醇修饰的 ManNAc 类似物（以及化学惰性的）的化学结构、动力学和剂量如何大小匹配的对照）调节目标 1a 中的细胞聚糖；评估细胞粘附和运动的变化目标 1b，并在目标 1c 中评估人类神经干细胞 (hNSC) 的分化。 2，我们将应用优化的模拟治疗条件来改善大鼠心脏的神经再生通过将 MGE 修饰移植到受伤动物中来建立脑损伤的逮捕 (CA) 模型我们将比较 hNSC。用我们新的硫醇修饰类似物进行治疗，并对 CA 后的功能恢复进行适当的控制在特定目标 3 中评估移植 hNSC 的存活、粘附、分布和迁移。我们将评估生化（Wnt 信号传导和钙粘蛋白参与）和细胞（组织浸润免疫）我们建议在脑损伤恢复中促进 MGE 的愈合作用（在 Aim 最后，在目标 3b 中，我们将通过质谱分析和使用来表征细胞范围的“糖位点”。具体而言，我们预计将通过糖生物信息学分析来识别未知的 MGE 生化介质。确定 MGE 在植入的 hNSC 以及反式作用宿主中的有益作用的新介质我们研究硫代类似物通过受体的复杂组合来调节 hNSC 的命运。对细胞信号传导和粘附的特定作用提供了一系列无法比拟的多效性治疗效果因此，我们的创新方法开辟了一条新途径。利用我们新的基于硫醇的 MGE 技术改善干细胞治疗。