Developing second generation SCID pig models: filling the gaps to improve translation of therapeutics in regenerative medicine

开发第二代 SCID 猪模型：填补空白，改善再生医学疗法的转化

• 批准号: 10559550
• 负责人: Christopher Tuggle
• 金额: $ 73.04万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10559550
• 关键词: Academia; Acceleration; Advisory Committees; Age; Age Months; Animal Model; Animals; B-Lymphocytes; Bone Marrow; Breeding; CD3 Antigens; Cancer cell line; Cardiac Myocytes; Cell Therapy; Cells; Clinic; Clinical Trials; Collaborations; Communication; Communities; Community Medicine; Data; Drug Implants; Engraftment; Family suidae; Fetus; Funding; Future; Generations; Genes; Genetic; Germ; Goals; Hematopoiesis; Hematopoietic stem cells; Human; Human Biology; Immune; Immune system; Immunologic Deficiency Syndromes; Immunophenotyping; In Vitro; Industry; Injections; Interleukin 2 Receptor Gamma; Knowledge; Link; Location; Longevity; Measurement; Methods; Modeling; Mus; Mutate; Natural Killer Cells; Neonatal; Operative Surgical Procedures; Outcomes Research; Phenotype; Physiology; Population; Pre-Clinical Model; Preclinical Testing; Preparation; Procedures; Protocols documentation; R24; Recommendation; Regenerative Medicine; Reporting; Research; Resources; Rodent Model; Safety; Severe Combined Immunodeficiency; Skin; Systems Development; T-Lymphocyte; Techniques; Testing; Therapeutic; Thymus Gland; Tissues; Translating; Translations; Transplantation; Xenograft Model; Xenograft procedure; artemis; biocontainment facility; cancer therapy; conditioning; efficacy testing; fetal; genetic resource; genome editing; human model; improved; in vivo; induced pluripotent stem cell; innovation; interest; intraperitoneal; male; mutant; neonatal human; novel; null mutation; pathogen; porcine model; postnatal; pre-clinical; safety study; safety testing; stem cell therapy; stem cells; tool; translational medicine; translational model; translational therapeutics; transplant model

Project Summary/Abstract: The promise of stem cell therapies has not been fully realized, due in part to a paucity of appropriate pre-clinical models. The pig is an excellent model of human biology, due to similarities of size, physiology, and genetics, and in some cases may be superior to rodent models, which often fail to provide data which effectively translates to human clinical trials. Thus, pig models that more accurately model humans are critically needed to improve research outcomes of regenerative medicine therapeutics and maximize safe translation to the clinic. As SCID pig models show promise in xenograft studies, this establishes further needs for humanized pigs as second-generation models and improved methods to routinely rear immunodeficient pigs for measurement of survival, safety, and potential efficacy of implanted therapeutic cells. We are focused on generating such new knowledge and demonstrating SCID pig use in preclinical regenerative medicine research. We have developed biocontainment facilities and protocols to raise Artemis (ART)-mutant SCID pigs. We have demonstrated this model's significance through successful ectopic engraftment with human induced pluripotent stem cells (iPSC) and human cancer cell lines, as well as survival of orthotopically transplanted human skin. To improve the model, we mutated the Interleukin 2 Receptor gamma gene (IL2RG) in a male ART null background. The ART null IL2RG null pig has the expected T-B-NK- phenotype, and we have successfully demonstrated partial human immune system development in these ART-IL2RG pigs. Thus we have taken pioneering steps to establish a pig SCID model, but improvements in xenograft safety/efficacy testing and humanization remain prerequisites to harness these research models for translational medicine. The specific objectives of this application are to a) validate SCID pig models for preclinical testing of cell and tissue xenografts, b) use multiple novel humanization approaches to determine the extent of human hematopoiesis in SCID pigs, and c) establish second-generation SCID pig model management protocols at multiple biocontainment facilities. The rationale for the proposed research is that our outbred SCID pig may more accurately reflect how proposed stem cell derived therapies will function in humans compared to mice. This project is innovative because we will use an integrated approach to combine research on the model's xenograft potential with research focused on protocols for improving the use of immunodeficient pigs, including humanization methods. We expect that the successful completion of this project will create genetic resources, data on xenograft and humanization rates, and associated animal procedures that will be highly desirable for SCID based modeling for research on the safety and efficacy of stem cell therapeutics. These unique resources are expected to have a significant impact in accelerating the translation of regenerative medicine research into the clinic.

项目概要/摘要：干细胞疗法的前景尚未完全实现，部分原因是 缺乏合适的临床前模型。猪是人类生物学的极好模型， 的大小，生理学和遗传学，并在某些情况下可能是上级啮齿动物模型，往往无法提供 这些数据可以有效地转化为人类临床试验。因此，猪模型更准确地模拟人类 迫切需要改善再生医学治疗的研究成果，并最大限度地提高安全性。 翻译到诊所随着SCID猪模型在异种移植研究中显示出希望，这建立了进一步的需求。 用于人源化猪作为第二代模型和常规饲养免疫缺陷猪的改进方法 用于测量植入的治疗性细胞的存活、安全性和潜在功效。我们专注于 产生了这样的新知识，并证明了SCID猪在临床前再生医学研究中的应用。 我们已经开发了生物隔离设施和协议，以提高阿耳忒弥斯（ART）突变SCID猪。我们 通过成功的异位移植， 多能干细胞（iPSC）和人类癌细胞系，以及原位移植的存活率 人类皮肤为了改进该模型，我们在男性ART中突变了白细胞介素2受体γ基因（IL 2 RG）。 空背景。ART无效IL 2 RG无效猪具有预期的T-B-NK-表型，并且我们已经成功地 在这些ART-IL 2 RG猪中证实了部分人免疫系统发育。因此，我们采取了 建立猪SCID模型的开创性步骤，但异种移植物安全性/有效性测试的改进， 人性化仍然是转化医学利用这些研究模式的先决条件。 本申请的具体目标是a）验证用于细胞临床前测试的SCID猪模型， 组织异种移植物，B）使用多种新的人源化方法来确定人源化的程度， c）建立第二代SCID猪模型管理方案， 多个生物防护设施这项研究的基本原理是，我们的远系繁殖的SCID猪可能更多地 准确地反映了与小鼠相比，所提出的干细胞衍生疗法将如何在人类中发挥作用。这 该项目是创新的，因为我们将使用一种综合的方法来联合收割机研究模型的异种移植 潜在的研究重点是改善免疫缺陷猪的使用方案，包括 人性化的方法我们期望该项目的成功完成将创造遗传资源， 异种移植和人源化率的数据，以及相关的动物程序，这些数据对于 基于SCID的干细胞治疗安全性和有效性研究建模。这些独特的资源 预计将对加速再生医学研究转化为 诊所

项目成果

Swine models for translational oncological research: an evolving landscape and regulatory considerations.

• DOI: 10.1007/s00335-021-09907-y
• 发表时间: 2022-03
• 期刊: Mammalian genome : official journal of the International Mammalian Genome Society
• 作者: Boettcher AN;Schachtschneider KM;Schook LB;Tuggle CK
• 通讯作者: Tuggle CK

Effect of ARTEMIS (DCLRE1C) deficiency and microinjection timing on editing efficiency during somatic cell nuclear transfer and in vitro fertilization using the CRISPR/Cas9 system.

• DOI: 10.1016/j.theriogenology.2021.04.003
• 发表时间: 2021-08
• 期刊: Theriogenology
• 影响因子: 2.8
• 作者: Li Y;Adur MK;Wang W;Schultz RB;Hale B;Wierson W;Charley SE;McGrail M;Essner J;Tuggle CK;Ross JW
• 通讯作者: Ross JW