Brain regeneration, an earthworm phenomenon: from molecular characterization to bioengineering a functional "brain in a dish"

大脑再生，一种蚯蚓现象：从分子表征到生物工程功能性“盘子里的大脑”

• 批准号: 1615891
• 金额: --
• 依托单位: King's College London
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-1615891
• 关键词: Brain; regeneration; earthworm; phenomenon; molecular

Strategic Research Priority: Bioscience for HealthAbstract The macro invertebrate earthworm is capable of fully regenerating its brain within 2-3 weeks of surgically removal. The molecular genetic basis of this biological phenomenon has not been studied in fine detail and the mechanistic drivers are largely elusive. The proposed project will redress this short-fall by identifying and characterizing key molecular transcripts and proteins involved in brain regeneration. These cellular and molecular cues will form the basis of the subsequent development of a bio-electrospray based approach to recreate a 3D functional earthworm brain ex vivo, i.e. in a dish.Project The proposed study brings together world-leading laboratories in earthworm molecular genetics (Prof Sturzenbaum, the supervisor at KCL), regenerative biology/medicine (Dr Jayasinghe, the supervisor at UCL) and earthworm immunology (Prof Laszlo Molnar, a project partner from the University of Pecs, Hungary). In concert, the two overarching points will be studied: 1) Molecular genetics will be applied to define what cells and molecules are involved during the development and regeneration of the earthworm brain. Specific molecular and cellular targets have been hypothesized to be involved in brain regeneration, namely the GABA-receptors, CAPA peptides, FVRIamides, Neuromacins, and coelomocytes (the immune cells), respectively. Furthermore the surgical removal and in vivo regeneration protocols are established and will be provided in liaison with Prof Molnár. However, many more drivers await discovery and characterization. Prof Sturzenbaum will supervise the target identification, which will be achieved by Subtractive Hybridization (e.g. using the PCR-Select cDNA Subtraction Kit, Clontech) and RNA-seq, the next-generation Whole Transcriptome Shotgun Sequencing (WTSS) approach. Identified biomarkers of regeneration will be validated by quantitative PCR (qPCR) in brains isolated at varying degrees of regeneration. 2) This above knowledge-base will be subsequently transferred to the UCL team. Individual cells and molecules will be mixed in a permutation / combination approach including a wide range of extracellular matrix (ECM) materials for bio-electrospraying. This will facilitate the subsequent formation of a compartmentalized bead containing all the constituents of the developing brain. The 3D proximity of the cells and molecules within these beads created at UCL will be used to study in detail the cellular and molecular cues at KCL, with the ultimate goal of developing, ex vivo, functional tissue of a regenerating brain.

战略研究重点：摘要大型无脊椎动物的大脑在手术切除后的2-3周内能够完全再生。这种生物现象的分子遗传学基础尚未得到详细研究，机械驱动因素在很大程度上是难以捉摸的。拟议的项目将通过识别和表征参与脑再生的关键分子转录本和蛋白质来弥补这一不足。这些细胞和分子线索将构成后续开发基于生物电喷雾的方法的基础，以在体外（即在培养皿中）重建3D功能性人脑。项目拟议的研究汇集了世界领先的生物分子遗传学实验室（Sturzenbaum教授，KCL的主管），再生生物学/医学（Jayasinghe博士，UCL的主管）和免疫学（Laszlo Molnar教授，匈牙利佩奇大学的项目合作伙伴）。在音乐会上，两个首要的点将被研究：1）分子遗传学将被应用于定义什么细胞和分子参与发育和再生的大脑。已经假设特定的分子和细胞靶点参与脑再生，即分别是GABA受体、CAPA肽、FVRI酰胺、Neuromacins和体腔细胞（免疫细胞）。此外，建立了手术切除和体内再生方案，并将与Molnár教授联系提供。然而，还有更多的驱动因素有待发现和表征。Sturzenbaum教授将监督目标识别，这将通过消减杂交（例如使用PCR-Select cDNA Subtraction Kit，Clontech）和RNA-seq（下一代全转录组鸟枪测序（WTSS）方法）实现。将在不同再生程度下分离的脑中通过定量PCR（qPCR）验证鉴定的再生生物标志物。 2)上述知识库随后将转移到UCL团队。单个细胞和分子将以排列/组合方法混合，包括用于生物电喷雾的各种细胞外基质（ECM）材料。这将促进随后形成含有发育中大脑的所有成分的区室化珠粒。在UCL创建的这些珠子内的细胞和分子的3D接近度将用于详细研究KCL的细胞和分子线索，最终目标是体外开发再生大脑的功能组织。