Miniature Swine Genetic Engineering and Characterization Core

核心C摘要 可获得的同种异体器官短缺是该领域最大的未得到满足的医疗需求。 今天的移植手术。过去，猪到灵长类器官移植的存活率有了显著的提高。 30年了，但不幸的是，仍不足以用于临床应用。这一领域的进一步进展很可能 依靠对猪源动物的遗传修改来克服与 灵长类动物受体，并允许诱导免疫耐受。这项建议的目标是 生产这样的猪，并确定有助于实现临床相关目标的额外基因修改 异种移植存活。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PRECISE CRISPR-CAS9系统的添加和编辑将被用于专门修改我们的近交猪 通过修改解决造血细胞因子的不相容问题(通过造血耐受 嵌合体)，以消除先前存在的抗体所识别的表位，这些表位是人类和狒狒共同的 受体(B4galNT2依赖)，并将人CD47定向到肾脏细胞，在其中可能发挥重要作用 保护作用(足细胞)。含有新修改的转基因猪将在 项目1和项目2：促进灵长类器官存活的有效性和诱导耐受的混合嵌合体 和人性化的老鼠模型。

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.