Novel humanized mouse model developed from cord blood CD34 positive HSC and autologous iPS cell derived thymus

由脐带血CD34阳性HSC和自体iPS细胞衍生胸腺开发的新型人源化小鼠模型

• 批准号: 9368151
• 负责人: Michael Allen Brehm
• 金额: $ 79.96万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9368151
• 关键词: Address; Animal Model; Animals; Antigen-Presenting Cells; Autologous; B-Lymphocytes; Bone Marrow; CD34 gene; CRISPR/Cas technology; Cell Differentiation process; Cell Line; Cell physiology; Clustered Regularly Interspaced Short Palindromic Repeats; Communicable Diseases; Communities; Complex; Development; Engraftment; Ensure; Epithelium; Fetal Tissues; Generations; Genes; Goals; HIV; Hematopoietic Cell Growth Factors; Hematopoietic stem cells; Human; Immune; Immune response; Immune system; Immunodeficient Mouse; Immunology; Institutes; Investigation; Knockout Mice; Liver; MHC Class I Genes; Manuscripts; Mission; Modeling; Mouse Strains; Mus; NIH Program Announcements; National Institute of Allergy and Infectious Disease; Natural Immunity; Natural regeneration; Nude Mice; Organ; Organoids; Outcome; Pharmacologic Substance; Pluripotent Stem Cells; Positioning Attribute; Pre-Clinical Model; Research; Specificity; Stem cells; Structure; T-Cell Development; T-Lymphocyte; Technology; The Jackson Laboratory; Thymic Tissue; Thymus Gland; Tissue Model; Tissue Procurements; Transgenic Mice; Transplanted tissue; Umbilical Cord Blood; United States National Institutes of Health; Work; adaptive immunity; base; cytokine; experimental study; fetal; human disease; human fetus tissue; human tissue; humanized mouse; immune function; immunodeficient mouse model; improved; in vivo; induced pluripotent stem cell; innovation; mouse model; multidisciplinary; new technology; next generation; novel; pre-clinical; reconstitution; response; tool; tumor

PROJECT SUMMARY/ABSTRACT Humanized mice, i.e., immunodeficient mice that support the development of human immune systems by engraftment with human hematopoietic stem cells (HSCs) have become increasingly important preclinical tools. However, HSC engraftment of NSG mice results in the development of H2-restricted human T cells that cannot interact appropriately with autologous human HLA-expressing, antigen-presenting cells and B cells in the periphery. NSG-HLA transgenic mice have been generated to address this, but the repertoire of HLA specificities is restricted to only those expressed and H2-restricted human T cells continue to be generated in the murine thymus. The most robust immune system in humanized NSG mice is created using fetal tissues, the BLT (bone marrow/liver/thymus) model. Use of the BLT model permits human T cell development on an autologous thymus and generates robust HLA-restricted human immune systems. However, some human T cells still develop in the murine thymus and are H2-restricted, confounding interpretation of the experiments. In addition, the use of human fetal tissue to create humanized mice is complicated by its limited availability and by concerns surrounding procurement of the tissues. To provide an alternative to fetal tissue-based models, we propose to utilize novel athymic NSG mouse strains developed by crosses of human cytokine trangenic NSG mice with our athymic NSG-Foxn1null (nude) mouse stock and by CRISPR/Cas targeting Foxn1 directly in our new NSG stocks expressing human hematopoietic growth factors. We will use umbilical cord blood-derived iPS cells to create human thymus structures that will support in vivo development of HLA-restricted human T cells derived from autologous cord blood HSCs. For this multi-PI, multi-disciplinary team project, we propose three Aims: 1) Generate and validate new models of immunodeficient mice that support optimal engraftment of HSCs and function of human thymic tissues in the absence of murine thymus; 2) Generate and validate human thymic epithelium and organoids generated from iPS cells; 3) Compare the human immune system generated in our new models with the BLT model. Our proposal takes advantage of powerful new technologies for creating new models of humanized mice, and builds on our >25 year track record for generating, validating, and sharing novel models of humanized mice. We believe that our innovative approaches, combined with our multi-disciplinary collaborative team will ensure the development of this much-needed preclinical model of humanized mice that does not rely on use of human fetal tissues. Furthermore, Dr. Shultz is at The Jackson Laboratory where these mouse models will be developed, facilitating their rapid distribution to the scientific community and uniquely positioning us to achieve the stated goals of this program announcement.

项目摘要/摘要 人源化小鼠，即通过免疫缺陷的小鼠，支持通过 与人类造血干细胞（HSC）植入已成为越来越重要的临床前 工具。然而，NSG小鼠的HSC植入导致H2限制的人T细胞的发展 无法与自体的人类HLA，抗原呈现细胞和B细胞适当相互作用 外围。已经生成了NSG-HLA转基因小鼠来解决这个问题，但是HLA的曲目 特异性仅限于仅表达和H2限制的人类T细胞的特异性。 鼠胸腺。人性化NSG小鼠中最健壮的免疫系统是使用胎儿组织创建的 BLT（骨髓/肝/胸腺）模型。 BLT模型的使用允许在 自体胸腺并产生强大的HLA限制性人体免疫系统。但是，一些人类 细胞在鼠胸腺中仍然发育，并受到H2限制性的限制，使实验的解释混淆。在 此外，使用人胎胎组织创建人源性小鼠的使用有限，并且 通过围绕组织采购的担忧。为了提供基于胎儿组织模型的替代方案， 我们建议利用人类细胞因子跨齿的杂交开发的新型无静脉NSG小鼠菌株 NSG小鼠与我们的nsg-foxn1null（裸）小鼠库存和CRISPR/CAS靶向FOXN1直接在 我们表达人类造血生长因素的新的NSG股票。我们将使用脐带鲜血 IPS细胞创建人类胸腺结构，这些结构将支持HLA受限的人T的体内发展 源自自体血液HSC的细胞。对于这个多pi的多学科团队项目，我们建议 三个目的：1）生成和验证免疫缺陷小鼠的新模型，以支持最佳植入 在没有鼠胸腺的情况下，人类胸腺组织的HSC和功能； 2）生成和验证人类 由IPS细胞产生的胸腺上皮和类器官； 3）比较生成的人免疫系统 在我们使用BLT模型的新模型中。我们的建议利用强大的新技术 创建新型人性化鼠标的新模型，并以我们的25年历史记录建立，以生成，验证， 并分享人性化小鼠的新型模型。我们认为，我们的创新方法与我们 多学科合作团队将确保开发这种急需的临床前模型 不依赖人类胎儿组织的人性化小鼠。此外，舒尔茨博士在杰克逊 将开发这些小鼠模型的实验室，促进它们的快速分布到科学 社区和独特的定位，以实现该计划公告的既定目标。