Miniature Swine Genetic Engineering and Characterization Core

小型猪基因工程和表征核心

• 批准号: 10328000
• 负责人: Robert J Hawley
• 金额: $ 41.31万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10328000
• 关键词: Address; Allogenic; Animal Sources; Animals; Antibodies; Binding; Biological Assay; Breeding; CD46 Antigen; CD47 gene; CRISPR/Cas technology; CSF2RA gene; Cell Transplantation; Cells; Chimerism; Colony-Stimulating Factor Receptors; Complement; Cytokine Receptors; Data; Databases; Dendritic Cells; Engraftment; Ensure; Epitopes; Euthanasia; Family suidae; Gene Expression; Gene Expression Profiling; Generations; Genes; Genetic; Genetic Engineering; Genotype; Goals; Granulocyte-Macrophage Colony-Stimulating Factor; Granulocyte-Macrophage Colony-Stimulating Factor Receptors; Hematopoietic; Human; IL3 Gene; IL3RA gene; Immune Tolerance; Immunoglobulin M; In Vitro; Inbreeding; Interleukin-3 Receptor; Kidney; Kidney Transplantation; Knock-out; Link; Medical; Miniature Swine; Modification; Molecular; Myelogenous; Organ Donor; Organ Survival; Organ Transplantation; Outcome; Papio; Phenotype; Play; Primates; Procedures; Production; Research Personnel; Role; Sampling; Schedule; Site; Standardization; System; Techniques; Testing; Time; Tissue Sample; Tissue Transplantation; Tissues; Transgenes; Transgenic Organisms; Transplantation; Work; Xenograft procedure; animal cloning; biobank; clinical application; clinically relevant; cytotoxicity; gain of function; gene function; genetic regulatory protein; humanized mouse; improved; interest; kidney cell; mouse model; nephrin; podocyte; preclinical development; progenitor; programs; promoter; protein expression; recombinase; research and development; transgene expression; transplant model

Core C Summary Shortage of available allogeneic donor organs represents the greatest unmet medical need in the field of transplantation today. The survival of pig-to-primate organ transplants has improved markedly over the past three decades but unfortunately remains insufficient for clinical application. Further progress in this field will likely depend upon genetic modifications of porcine source animals both to overcome molecular incompatibilities with primate recipients and to allow the induction of immunological tolerance. The objectives of this proposal are to produce such pigs and to identify additional genetic modifications that will help attain the goal of clinically relevant xenograft survival. Advanced genetic engineering techniques, including site-specific recombinase-driven gene additions and edits using the precision CRISPR-Cas9 system, will be used to specifically modify our inbred swine with modifications addressing incompatibilities in hematopoietic cytokines (tolerance through hematopoietic chimerism), to eliminate epitopes recognized by pre-existing antibodies common to human and baboon recipients (B4galNT2-dependent) and to direct human CD47 to kidney cells in which it may play an important protective role (podocytes). Genetically modified pigs incorporating the new modifications will be tested in Projects 1 and 2 for efficacy in promoting organ survival and mixed chimerism for tolerance induction in primate and humanized mouse models.

核心C摘要 可用的同种异体供体器官短缺是移植领域最大的未满足的医疗需求。 移植今天猪到灵长类动物器官移植的存活率在过去有了显著提高 30年，但不幸的是，仍然不足以用于临床应用。这一领域的进一步进展将可能 依赖于猪源动物的遗传修饰， 灵长类受体，并允许诱导免疫耐受。本提案的目标是 生产这样的猪，并确定额外的遗传修饰，这将有助于实现临床相关的目标， 异种移植物存活率先进的基因工程技术，包括位点特异性重组酶驱动的基因 使用精确的CRISPR-Cas9系统进行添加和编辑，将用于专门修饰我们的近交系猪。 通过修改解决了造血细胞因子的不相容性（通过造血细胞因子的耐受性） 嵌合），以消除由人和狒狒共有的预先存在的抗体识别的表位 受体（B4 galNT 2依赖性），并将人CD 47导向肾细胞，在肾细胞中它可能发挥重要作用。 保护作用（足细胞）。含有新基因改造的转基因猪将在 项目1和2在灵长类动物中促进器官存活和诱导耐受的混合嵌合体的功效 和人源化小鼠模型。