Developing preclinical xenograft models in zebrafish.

在斑马鱼中开发临床前异种移植模型。

基本信息

批准号：
10578692
负责人：
David Michael Langenau
金额：
$ 79.46万
依托单位：
MASSACHUSETTS GENERAL HOSPITAL
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：
2022-03-01 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10578692
关键词：
Ablation Address Adult Animal Model Animals Apoptosis B-Lymphocytes Biological Biological Immunotherapy Blood Blood Cells CD34 gene Cell Cycle Cell Death Cell Transplantation Cell division Cell physiology Cells Clinical Combination Drug Therapy Communities DNA Damage Development Embryo Engraftment Genotype Goals Growth Human Image Imaging Device Immune Immunotherapy Knock-in Label Laboratories Macrophage Malignant Neoplasms Methods Mission Modeling Mus 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 Neoplasm Metastasis Optics Pharmaceutical Preparations Pharmacodynamics Pluripotent Stem Cells Poly(ADP-ribose) Polymerase Inhibitor Positioning Attribute Pre-Clinical Model Proliferating Protocols documentation Radiation Recording of previous events Recovery Research Resolution Rhabdomyosarcoma Self Assessment Study models T-Lymphocyte Testing Therapeutic Therapy Evaluation Transgenic Organisms Transplantation Umbilical Cord Blood United States National Institutes of Health Visualization Work Xenograft Model Xenograft procedure Zebrafish cancer cell cell behavior cell growth cell type cellular imaging chimeric antigen receptor T cells cost cytokine drug sensitivity fluorescence imaging human disease human imaging human tissue imaging approach imaging platform in vivo in vivo Model induced pluripotent stem cell malignant muscle neoplasm mutant neoplastic cell next generation novel novel therapeutics patient derived xenograft model pre-clinical pre-clinical assessment pre-clinical therapy radiation response real-time images regenerative biology regenerative cell regenerative tissue response self-renewal stem stem cell self renewal stem cells success temozolomide tool translational impact transplant model treatment response tumor tumor growth

项目摘要

PROJECT SUMMARY Xenograft cell transplantation has transformed our understanding of human disease and has been used extensively to assess regeneration, stem cell self-renewal, and cancer. Yet, mouse xenograft studies are expensive and not easily amenable to imaging engraftment at single cell resolution. By contrast, zebrafish are inexpensive, can be reared in large numbers, and are capable of real-time imaging of fluorescent-labeled cells at single cell resolution. Our group has recently pioneered the use of adult immune-deficient zebrafish for xenograft transplantation of human cancers and blood cells when reared at 37OC. Despite these successes, more needs to be done to develop the next generation of immune compromised zebrafish for long-term xenograft cell transplantation studies. The long-term goal of our work is to develop a universal zebrafish transplantation model to engraft of a wide array of human regenerative and cancer cell types. The overall objective is i) to develop new immune deficient zebrafish models for optimized xenograft engraftment of human cancer, embryonic and induced-pluripotent stem cells (ES and iPSCs), and blood cells and ii) provide much needed tools, methods, and cell biological readouts to directly assess pharmacodynamic responses to radiation, drugs, and cell biological immunotherapies in vivo. The rationale for our research is that zebrafish blood development is highly conserved and that developing zebrafish transplantation models will provide new tools to rapidly assess preclinical therapies in vivo and at single cell resolution. Aim 1 will develop compound mutant and transgenic zebrafish for optimized xenograft cell transplantation. We will develop new models that lack T, B, NK, and macrophage cell function and that transgenically express human cytokines to support the growth of human blood. We will also generate knock-in “genotype-less rag2∆/∆, il2rga−/− zebrafish” to increase throughput in identifying double homozygous mutant animals. Aim 2 will test these models for enhanced engraftment of human cancers, ES, iPSCs, and blood cells. This work is important, because it will provide novel models and experimental protocols to engraft a wide array of regenerative cell types. Aim 3 will dynamically visualize xenograft single cell responses to radiation, combination drug therapies, and immunotherapy in preclinical modeling studies. This work will provide much needed cell biological readouts to directly assess pharmacodynamic responses at single cell resolution across a wide array of therapies. These same imaging tools and approaches can be used in many xenograft models – including patient-derived xenografts (PDXs), ES/IPSCs, and blood. Our work is significant because it will develop the next generation of low-cost, high throughput cell transplantation models that allow direct visualization of engrafted cell behaviors in the context of preclinical therapies. This work will have a positive translational impact by developing preclinical animal models that efficiently engraft a wide array of human tissues. Such broad reaching applications for immune compromised zebrafish spans the mission of many NIH institutes.

项目概要异种移植细胞移植改变了我们对人类疾病的理解并已被用于通常，小鼠异种移植研究是为了评估再生、干细胞自我更新和癌症。相比之下，斑马鱼价格昂贵且不易以单细胞分辨率成像。价格便宜，可以大量培养，并且能够对荧光标记的细胞进行实时成像我们的小组最近率先使用成年免疫缺陷斑马鱼进行单细胞分辨率的研究。尽管取得了这些成功，但在 37OC 下培养的人类癌症和血细胞的异种移植。需要做更多的工作来长期开发下一代免疫受损斑马鱼我们工作的长期目标是开发通用的斑马鱼。移植模型可移植多种人类再生细胞和癌细胞类型。目标是 i) 开发新的免疫缺陷斑马鱼模型，以优化人类异种移植癌症、胚胎干细胞和诱导多能干细胞（ES 和 iPSC）以及血细胞和 ii) 提供许多需要工具、方法和细胞生物学读数来直接评估药效学反应我们研究的基本原理是斑马鱼体内的辐射、药物和细胞生物免疫疗法。血液发育高度保守，开发斑马鱼移植模型将提供新的目标 1 将开发化合物，以在体内和单细胞分辨率下快速评估临床前疗法。我们将开发用于优化异种移植细胞移植的突变和转基因斑马鱼。缺乏 T、B、NK 和巨噬细胞功能，并且转基因表达人类细胞因子以支持我们还将生成敲入“无基因型 rag2Δ/Δ, il2rga−/− 斑马鱼”以增加目标 2 将测试这些模型的增强能力。人类癌症、ES、iPSC 和血细胞的植入这项工作很重要，因为它将提供。 Aim 3 将提供移植多种再生细胞类型的新模型和实验方案。动态可视化异种移植单细胞对放射、联合药物治疗的反应，以及这项工作将为临床前模型研究中的免疫治疗提供急需的细胞生物学读数。以单细胞分辨率直接评估多种疗法的药效反应。相同的成像工具和方法可用于许多异种移植模型 - 包括患者来源的模型异种移植物 (PDX)、ES/IPSC 和血液我们的工作意义重大，因为它将开发下一代。低成本、高通量的细胞移植模型，可以直接观察雕刻的细胞这项工作将产生积极的转化影响。开发可有效移植多种人体组织的临床前动物模型。针对免疫受损斑马鱼的应用跨越了许多 NIH 机构的使命。