Danionella cerebrum as a transparent vertebrate adult model for studying immune-related biological processes and diseases

大脑丹尼奥菌作为透明脊椎动物成年模型，用于研究免疫相关的生物过程和疾病

• 批准号: 10665376
• 负责人: Pui Ying Lam
• 金额: $ 23.4万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10665376
• 关键词: Acceleration; Adult; Affect; Aging; Animal Model; Animals; Behavioral; Biological Models; Biological Process; Biology; Brain; Calcium; Cell physiology; Cells; Central Nervous System; Cephalic; Cerebrum; Characteristics; Collection; Communities; Complex; DNA; Data; Data Set; Development; Disease; Disease Progression; Disease model; Dorsal; Endothelial Cells; Environment; Equipment; Event; Female; Future; Gene Transfer Techniques; Genetic Engineering; Goals; Heart; Heterogeneity; Image; Imaging Techniques; Immune; Immune response; Immune system; Immunological Models; Immunology; Inflammaging; Infrastructure; Injury; Institution; Kidney; Knowledge; Label; Larva; Learning; Length; Light; Liver; Macrophage; Malignant Neoplasms; Measures; Mechanics; Memory; Methods; Microglia; Microscopy; Modeling; Molecular; Mus; Names; Natural Immunity; Natural regeneration; Nature; Nerve Regeneration; Nervous System; Neurodegenerative Disorders; Optics; Organ; Physiological; Plasmids; Play; Procedures; Process; Reagent; Regenerative capacity; Regenerative research; Reporter; Research; Resolution; Role; Sex Differences; Study models; Systems Biology; Tail; Techniques; Testing; Therapeutic; Tissue imaging; Tissues; Transgenic Animals; Transgenic Organisms; Vascular System; Vertebrates; Visualization; Work; Zebrafish; adaptive immunity; addiction; age related; aged; awake; body cavity; body system; bone; cell type; comparative; cranium; experience; experimental study; genome editing; high resolution imaging; human disease; imaging modality; imaging study; immune cell infiltrate; improved; in vivo; in vivo Model; in vivo imaging; infancy; intravital imaging; male; model organism; mutant; neutrophil; novel; optogenetics; regenerative; response; response to injury; screening; sex; small molecule; teleost; tissue injury; tissue regeneration; tool; transcriptomics; two-photon; wound healing; young adult

An optically accessible adult vertebrate model is needed to study dynamic cellular processes in vivo. We aim to develop and characterize Danionella cerebrum, a teleost related to zebrafish that remains transparent into adulthood, for studying immune-related biological processes and their impact on disease progression in multiple organs. A transparent adult model is particularly important for live imaging studies of immune related processes due to the dynamic and complex nature of immune cell heterogeneity and plasticity, the involvement of infiltrating immune cells during disease progression, as well as confounding factors related to aging. D. cerebrum, previously named as D. translucida, was first described by the Judkewitz group in 2018 for brain-related studies. This genetically tractable model is small, adults reach a length of approximately 12 mm in length, and is optical transparency even into adulthood. Established transgenesis and genome editing methods used in zebrafish have showed to be effective in D. cerebrum. These characteristics make D. cerebrum an attractive model for noninvasive in vivo visualization of dynamic and complex cellular events in a physiologically relevant setting. This study aims to build the infrastructure required to use D. cerebrum as a systems biology model organism for studying immune-related biological processes and their impact on disease progression. Our proof of principle experiments have successfully generated transgenic lines with fluorescently labelled innate immune cells such as neutrophils and macrophages, and the endothelial cells of the vasculature. We propose to expand our collection of transgenic D. cerebrum lines by fluorescently labeling multiple organs such as the brain, heart, liver, and kidney. In combination, these transgenic lines will allow us to visualize the complex interplay between immune cells and various organs throughout development, injury and disease. We will also develop a macrophage/microglia fluorescent activity reporter line which will allow us to observe and measure dynamic state changes displayed by immune cells in a physiologically relevant environment using non-invasive in vivo imaging. To facilitate our studies, we will develop methods and equipment for longitudinal live imaging of awake adult D. cerebrum. Immune cell functions play important roles in wound healing and regeneration. Zebrafish are known to have great regeneration capacity in different organs, however, it is not known if D. cerebrum can regenerate. Our preliminary work suggests that adult D. cerebrum can regenerate its tail fin but this regenerative capacity decreased in aged animal. Our proposed study will investigate if D. cerebrum regenerates its central nervous system during different developmental stages, its corresponding immune cell response and the transcriptomic changes that accompany these events. We will also perform interspecies comparative analysis using available data from zebrafish regeneration studies to explore key players in modulating regeneration capacity. This proposal describes strategies that will build the infrastructure required to use D. cerebrum for immunology related studies in vivo. The tools, techniques and knowledge built for the D. cerebrum immune model will provide an important springboard for carrying out future focused analyses on immune-related processes that include, but by no means are limited to, inflammaging, wound healing/regeneration, cancer, and neurodegenerative diseases.

一个光学可访问的成年脊椎动物模型是需要在体内研究动态细胞过程。我们的目标是开发 并描述大脑丹尼翁氏菌（Danionella cerebrum）的特征，这是一种与斑马鱼相关的硬骨鱼，在成年后仍然透明，以供研究 免疫相关的生物过程及其对多器官疾病进展的影响。一个透明的成人模型 对于免疫相关过程的活体成像研究尤其重要， 免疫细胞的异质性和可塑性，疾病进展过程中浸润免疫细胞的参与，以及 作为与衰老相关的混杂因素。D. cerebrum，以前命名为D.最早被描述为 Judkewitz小组在2018年进行了大脑相关研究。这种基因上易于处理的模型很小，成年人达到一个长度 长约12 mm，即使成年后也是光学透明的。建立转基因和基因组 在斑马鱼中使用的编辑方法已经证明在D.大脑这些特点使D.大脑 在生理相关的非侵入性体内可视化动态和复杂的细胞事件的有吸引力的模型 设置.本研究旨在建立使用D.大脑作为系统生物学模式生物， 研究免疫相关的生物过程及其对疾病进展的影响。我们的原理实验证明 已经成功地产生了具有荧光标记的先天免疫细胞如嗜中性粒细胞的转基因系， 巨噬细胞和脉管系统的内皮细胞。我们建议扩大我们的收集转基因D。大脑 通过荧光标记多个器官，如大脑、心脏、肝脏和肾脏，这些转基因的组合， 线将使我们能够可视化整个发育过程中免疫细胞和各种器官之间的复杂相互作用， 伤害和疾病。我们还将开发巨噬细胞/小胶质细胞荧光活性报告细胞系，这将使我们能够 观察和测量免疫细胞在生理相关环境中显示的动态变化， 非侵入性体内成像。为了便于我们的研究，我们将开发纵向现场的方法和设备， 清醒成人的成像D.大脑免疫细胞功能在伤口愈合和再生中起重要作用。 已知斑马鱼在不同器官中具有很强的再生能力，然而，尚不清楚D。大脑罐 再生我们的初步工作表明，成年D.大脑可以再生尾鳍，但这种再生能力 老年动物减少。我们的研究将探讨如果D。大脑再生其中枢神经系统， 不同的发育阶段，其相应的免疫细胞反应和转录组的变化，伴随 这些事件。我们还将利用斑马鱼再生的可用数据进行物种间比较分析 研究探索调节再生能力的关键参与者。该提案描述了将建立 基础设施需要使用D。用于体内免疫学相关研究。工具、技术和知识 为D而建。大脑免疫模型将为未来开展重点分析提供一个重要的跳板， 免疫相关过程包括但不限于炎症、伤口愈合/再生、癌症 和神经退行性疾病。