Immune Compromised Zebrafish for Cell Transplantation

用于细胞移植的免疫受损斑马鱼

• 批准号: 10454455
• 负责人: David Michael Langenau
• 金额: $ 17.12万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454455
• 关键词: Adult; Allogenic; Allografting; Animal Model; Animals; B-Lymphocytes; Blood; Blood Cells; CD34 gene; Cancer cell line; Cell Transplantation; Cell physiology; Cells; Chemicals; Communication; Communities; Community Medicine; Defect; Development; Eating; Engraftment; Ensure; Feedback; Fishes; Generations; Genes; Genome engineering; Germ; Goals; Gold; Growth; Heart; Homologous Transplantation; Human; Immune; Institutes; Label; Laboratories; Liver; Malignant Neoplasms; Mature T-Lymphocyte; Methods; Mission; Modeling; Monoclonal Antibody R24; Mus; Muscle Cells; NK-lysin; National Heart, Lung, and Blood Institute; National Institute of Arthritis and Musculoskeletal and Skin Diseases; National Institute of Child Health and Human Development; National Institute of Diabetes and Digestive and Kidney Diseases; National Institute of Neurological Disorders and Stroke; Natural Killer Cells; Natural regeneration; Optics; Pathway interactions; Positioning Attribute; Protocols documentation; Regenerative Medicine; Research; Resolution; Resources; Signal Transduction; System; T-Lymphocyte; Techniques; Time; Tissue Microarray; Tissues; Training; Transgenic Organisms; Transplantation; Umbilical Cord Blood; United States National Institutes of Health; Visualization; Water; Work; Xenograft procedure; Zebrafish; cancer cell; cancer stem cell; cell behavior; cell growth; cell type; chemical genetics; cost; cytokine; cytotoxic; drug sensitivity; experimental study; fluorescence imaging; genetic approach; genetic regulatory protein; human disease; human tissue; in vivo; induced pluripotent stem cell; loss of function mutation; mouse model; mutant; neovascularization; next generation; novel; pre-clinical; real-time images; regenerative; regenerative biology; regenerative cell; regenerative tissue; self-renewal; stem cell function; stem cell self renewal; stem cells; success; tool; translational impact; transplant model; treatment response; webinar

Cell transplantation into immune compromised mice has transformed our understanding of human disease and has been used extensively to assess regeneration, stem cell self-renewal, and cancer in the xenograft transplantation setting. Despite their great utility, mouse models are not amenable to large-scale studies due to high husbandry costs and do not easily facilitate direct visualization of engrafted cells at single cell resolution. By contrast, zebrafish are inexpensive, can be reared in large numbers, and are amenable to large-scale chemical genetic approaches where compounds can be added directly to the water. Moreover, optically-clear immune-deficient zebrafish strains have permitted large-scale cell transplantation studies to dynamically image fluorescent-labeled cells at single cell resolution. Despite these successes, more needs to be done to develop immune compromised zebrafish as a robust and long-term xenograft cell transplantation model. The long-term goal of this application is to develop a universal zebrafish transplantation model for engrafting a wide array of regenerative and cancer cell types from zebrafish, mouse, and human. The overall objective of this application is to provide new immune deficient zebrafish models for optimized allograft engraftment of regenerative tissues and xenograft engraftment of human cancer, ES, iPS, and CD34+ cord blood cells. The rationale for our research is that zebrafish blood development is highly conserved and that developing zebrafish transplantation models has already led to unique understanding of regenerative stem cell processes and dynamic visualization of new cell behaviors that drive cell growth. Aim 1 will develop compound mutant and humanized transgenic zebrafish for optimized cell transplantation. We will develop new models that lack all T, B, and NK cells, including mutants in the recently identified NK-lysin expressing cytotoxic blood cells and full loss-of-function mutations in the rag2 gene, which is required for mature T and B cell function. We will also generate humanized zebrafish that transgenically express factors that support elevated growth of human cells, including the human “don’t eat me” signal inhibitory regulatory protein alpha (SIRPa) and human cytokines. Aim 2 will utilize these models for assessing orthotopic and xenograft engraftment, identifying lines that have superior, long-term engraftment of human cancer cell lines, ES and iPS cells, and CD34+ cord blood cells. Aim 3 will refine a system for global distribution and rapid dissemination of mutant lines to the zebrafish, stem cell, and regenerative medicine community. Our work is significant because it will develop a much-needed resource for the community, facilitating the next generation of low-cost, high throughput cell transplantation models to engraft a wide array of regenerative cell types. This work is expected to have a positive translational impact by developing pre-clinical animal models that facilitate direct visualization of engrafted cells at reduced cost and allow chemical genetic approaches to uncover pathways associated with regeneration and stem cell function. Such broad reaching applications for immune compromised zebrafish spans the mission of many NIH institutes.

细胞移植到免疫受损的小鼠中改变了我们对人类疾病和 已广泛用于评估异种移植物中的再生、干细胞自我更新和癌症 尽管小鼠模型具有很大的实用性，但由于其不适合大规模研究。 饲养成本高，并且不容易促进以单细胞分辨率直接观察雕刻细胞。 相比之下，斑马鱼价格便宜，可以大量饲养，并且适合大规模饲养 此外，化学遗传方法可以将化合物直接添加到水中。 免疫缺陷斑马鱼品系允许大规模细胞移植研究动态成像 尽管取得了这些成功，但仍需要做更多的工作来开发单细胞分辨率的荧光标记细胞。 免疫受损的斑马鱼作为稳健且长期的异种移植细胞移植模型。 该应用程序的目标是开发一种通用的斑马鱼移植模型，用于移植多种斑马鱼 来自斑马鱼、小鼠和人类的再生细胞和癌细胞类型。 旨在提供新的免疫缺陷斑马鱼模型，用于优化再生组织的同种异体移植 人类癌症、ES、iPS 和 CD34+ 脐带血细胞的异种移植。 研究表明，斑马鱼血液发育高度保守，并且正在开发斑马鱼移植 模型已经使人们对再生干细胞过程和动态可视化有了独特的理解 目标 1 将开发复合突变体和人源化转基因。 我们将开发缺乏所有 T、B 和 NK 细胞的新模型。 包括最近发现的表达细胞毒性血细胞 NK 溶素的突变体和完全功能丧失 我们还将生成 rag2 基因的突变，这是成熟 T 和 B 细胞功能所必需的。 人源化斑马鱼转基因表达支持人类细胞生长加快的因子，包括 人类“别吃我”信号抑制调节蛋白 α (SIRPa) 和人类细胞因子 Aim 2 会。 利用这些模型来评估原位和异种移植物的植入，识别具有优越、 人类癌细胞系、ES 和 iPS 细胞以及 CD34+ 脐带血细胞 Aim 3 的长期植入。 完善一个系统，用于将突变系快速传播到斑马鱼、干细胞和 我们的工作意义重大，因为它将为再生医学界开发急需的资源。 社区，促进下一代低成本、高通量细胞移植模型 这项工作预计将产生积极的转化影响。 开发临床前动物模型，有助于以较低的成本直接观察雕刻细胞 允许化学遗传学方法揭示与再生和干细胞功能相关的途径。 免疫受损斑马鱼的如此广泛的应用涵盖了许多 NIH 研究所的使命。

项目成果

Targeting prostate-specific membrane antigen for personalized therapies in prostate cancer: morphologic and molecular backgrounds and future promises.

针对前列腺特异性膜抗原进行前列腺癌个性化治疗：形态学和分子背景以及未来的前景。

• DOI: 10.1021/jm9505977
• 发表时间: 2014-10-01
• 期刊: Journal of biological regulators and homeostatic agents
• 影响因子: 3.2
• 作者: M. Santoni;M. Scarpelli;R. Mazzucchelli;A. López;Liang Cheng;S. Cascinu;R. Montironi
• 通讯作者: R. Montironi

Cell of origin dictates aggression and stem cell number in acute lymphoblastic leukemia.

起源细胞决定了急性淋巴细胞白血病的攻击性和干细胞数量。

• 发表时间: 2018-08
• 影响因子: 11.4
• 作者: Garcia, Elaine G;Iyer, Sowmya;Garcia, Sara P;Loontiens, Siebe;Sadreyev, Ruslan I;Speleman, Frank;Langenau, David M
• 通讯作者: Langenau, David M

Identification and characterization of T reg-like cells in zebrafish.

斑马鱼中 T reg 样细胞的鉴定和表征。

• 发表时间: 2017-12-04
• 作者: Kasheta, Melissa;Painter, Corrie A;Moore, Finola E;Lobbardi, Riadh;Bryll, Alysia;Freiman, Eli;Stachura, David;Rogers, Arlin B;Houvras, Yariv;Langenau, David M;Ceol, Craig J
• 通讯作者: Ceol, Craig J

The macrophage-expressed gene (mpeg) 1 identifies a subpopulation of B cells in the adult zebrafish.

巨噬细胞表达基因 (mpeg) 1 识别成年斑马鱼中的 B 细胞亚群。

• 发表时间: 2020
• 影响因子: 5.5
• 作者: Ferrero, Giuliano;Gomez, Etienne;Lyer, Sowmya;Rovira, Mireia;Miserocchi, Magali;Langenau, David M;Bertrand, Julien Y;Wittamer, Valérie
• 通讯作者: Wittamer, Valérie

Evolutionary origin of cAMP-based chemoattraction in the social amoebae.

社会变形虫中基于 cAMP 的化学吸引的进化起源。

• 发表时间: 2005-05-03
• 影响因子: 11.1
• 作者: Alvarez;Rozen, Daniel E;Ritchie, Allyson V;Fouquet, Celine;Baldauf, Sandra L;Schaap, Pauline
• 通讯作者: Schaap, Pauline