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.

项目摘要异种移植细胞移植已改变了我们对人类疾病的理解，并已被使用广泛评估再生，干细胞自我更新和癌症。然而，鼠标的元素研究是昂贵，不容易想象单细胞分辨率的植入。相比之下，斑马鱼是便宜的，可以大量真实，并且能够对荧光标记的细胞进行实时成像在单细胞分辨率下。我们的小组最近开创了成人免疫缺陷斑马鱼的使用 37oC饲养时，人类癌和血细胞的异种移植移植。尽管取得了这些成功，需要做更多的事情，以发展下一代免疫折衷的斑马鱼长期异种移植细胞移植研究。我们工作的长期目标是开发普遍的斑马鱼移植模型植入了各种各样的人类再生和癌细胞类型。总体目的是i）开发新的免疫缺陷斑马鱼模型，以优化人类的异种移植癌症，胚胎和诱导的血清干细胞（ES和IPSC），血细胞和II）提供了很多需要的工具，方法和细胞生物学读数，以直接评估对药效的反应辐射，药物和细胞生物学免疫疗法在体内。我们研究的理由是斑马鱼血液发育是高度保守的，开发斑马鱼移植模型将提供新的快速评估体内和单细胞分辨率的临床前疗法的工具。 AIM 1将发展复合突变体和转基因斑马鱼，用于优化的Xenographic细胞移植。我们将开发新的模型缺乏T，B，NK和巨噬细胞功能，并且翻译表达人类细胞因子以支持人血的生长。我们还将生成“无基因型rag2Δ/∆，il2rga - / - 斑马鱼”以增加敲门识别双重纯合突变动物的吞吐量。 AIM 2将测试这些模型以增强人类癌，ES，IPSC和血细胞的植入。这项工作很重要，因为它将提供新型模型和实验方案，以植入各种再生细胞类型。目标3意志动态地可视化对辐射，联合药物疗法和临床前建模研究中的免疫疗法。这项工作将为急需的细胞生物读数提供直接评估各种疗法的单细胞分辨率下的药效反应。这些相同的成像工具和方法可以在许多异种移植模型中使用 - 包括患者来源异种移植物（PDXS），ES/IPSC和血液。我们的工作很重要，因为它将发展下一代低成本的高吞吐量细胞移植模型，可直接可视化植入的细胞临床前疗法的行为。这项工作将对开发有效植入各种人体组织的临床前动物模型。如此广泛杀伤斑马鱼的申请跨越了许多NIH机构的任务。