Miniature Swine Genetic Engineering and Characterization Core

小型猪基因工程和表征核心

基本信息

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
项目类别：
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

项目摘要

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摘要缺乏可用的同种异体供体器官代表了该领域最大的未满足医疗需求今天的移植。过去，猪到顶峰的器官移植的生存率显着提高三十年，但不幸的是，在临床应用中仍然不足。该领域的进一步进展可能会依赖猪源动物的遗传修饰都克服了分子不相容性灵长类动物接受者，并允许诱导免疫学耐受性。该提议的目标是生产此类猪并确定其他遗传修饰，以帮助实现临床相关的目标异种移植生存。先进的基因工程技术，包括特异性重组酶驱动的基因使用Precision CRISPR-CAS9系统的添加和编辑将用于专门修改我们的近交猪通过解决造血细胞因子中不兼容的修改（通过造血的耐受性嵌合），消除通过人和狒狒共有的抗体所识别的表位接受者（依赖B4GALNT2）并将人CD47引导到可能发挥重要作用的肾脏细胞保护作用（足细胞）。结合新修饰的转基因猪将在项目1和2，用于促进器官生存和混合嵌合的功效，以使灵长类动物的耐受性诱导和人源化的小鼠模型。