TRANSGENIC PIG WHICH SIMULTANEOUSLY EXPRESSES HO-1 GENE AND TNFR1-FC GENE, AND COMPRISES KNOCKED-OUT GGTA1 GENE, AND USE THEREOF

• Main Authors: LEE, Eun Mi, LEE, Byeong Chun, KIM, Su Jin, KIM, Geon A, LEE, Sang Hoon, AHN, Cu Rie, CHO, Bum Rae, HONG, Hye Jin, YEOM, Su Cheong, HWANG, In Chang
• Format: Patent
• Language: English; French; Korean
• Published: 18.05.2017
• Subjects: AGRICULTURE; ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; FORESTRY; HUMAN NECESSITIES; HUNTING; NEW BREEDS OF ANIMALS; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; TRAPPING

The present invention relates to a transgenic pig which simultaneously expresses a heme oxygenase-1 (HO-1) gene and a tumor necrosis factor receptor 1-Fc (TNFR1-Fc) gene and comprises a knocked-out α-1,3-galactosyltransferase (GGTA1) gene and in which an immune rejection response is inhibited upon xenotransplantation, and a method for preparing the same. The transgenic pig which simultaneously expresses human HO-1 and TNFR1-Fc fusion genes and comprises a knocked-out GGTA1 gene, according to the present invention, can reduce oxidative stress upon organ separation and in vitro culturing by the antioxidative action, cell protection functions, etc. of the HO-1 gene, can reduce a TNF-α mediated inflammatory response occurring during an initial transplant period by the expression of TNFR1-Fc, and also can inhibit maturation of dendritic cells and control the proliferation and activation of T-cells to reduce acute vascular rejection, thereby improving the initial engraftment of the transplanted organ. Furthermore, a hyperacute immune rejection is inhibited by GGTA1 whereby the survival rate of the transplanted organ is increased. Therefore, it is possible to produce an organ in which an immune rejection response is inhibited upon heterograft using the transgenic pig. La présente invention concerne un porc transgénique qui exprime simultanément un gène d'hème oxygénase-1 (HO-1) et un gène de récepteur de facteur de nécrose tumorale 1-Fc (TNFR1-Fc), et comprend un gène de α-1,3-galactosyltransférase (GGTA1) inactivé et dans lequel une réponse de rejet immunitaire est inhibée lors d'une xénogreffe, et son procédé de préparation. Le porc transgénique qui exprime simultanément des gènes de fusion HO-1 et TNFR1-Fc humains et comprend un gène GGTA1 inactivé, selon la présente invention, peut réduire le stress oxydatif lors d'une séparation d'organe et d'une culture in vitro par l'action antioxydante, des fonctions de protection cellulaire, etc., du gène HO-1, peut réduire une réponse inflammatoire à médiation par TNF-α se produisant pendant une période de greffe initiale par l'expression de TNFR1-Fc, et peut également inhiber la maturation de cellules dendritiques et lutter contre la prolifération et l'activation de lymphocytes T pour réduire un rejet vasculaire aigu, en améliorant ainsi la greffe initiale de l'organe greffé. En outre, un rejet immunitaire hyperaigu est inhibé par GGTA1, le taux de survie de l'organe greffé étant augmenté. Par conséquent, il est possible de produire un organe dans lequel une réponse de rejet immunitaire est inhibée lors d'une hétérogreffe au moyen du porc transgénique. 본 발명은 HO-1 (Heme oxygenase-1) 유전자 및 TNFR1-Fc (Tumor necrosis factor receptor 1-Fc) 유전자를 동시에 발현하며 GGTA1 (α-1,3-galactosyltransferase) 유전자가 넉아웃 (knock-out)된, 이종 장기 이식 시 면역 거부 반응이 억제된 형질전환 돼지, 및 이의 제조방법에 관한 것이다. 본 발명의 인간 HO-1과 TNFR1-Fc 융합 유전자를 동시 발현하며 GGTA1 유전자가 넉아웃된 형질전환 돼지는 HO-1 유전자의 항산화 작용 및 세포 보호 기능 등에 의해 장기 분리 및 체외 배양 시에 산화적 스트레스를 감소시킬 수 있고, TNFR1-Fc 발현에 의해 이식 초기에 일어나는 TNF-α 매개의 염증 반응도 감소시킬 수 있으며, 또한 수지상 세포의 성숙을 억제하고 T 세포의 증식과 활성화를 조절하여 급성 혈관계 거부 반응을 감소시켜 이식된 장기의 초기 생착을 촉진할 수 있다. 나아가, GGTA1에 의한 초급성 면역 거부반응을 억제함으로써 이식된 장기의 생존율을 증가시킬 수 있다. 이에, 상기 형질전환 돼지를 이용하여 이종 이식 시 면역 거부 반응이 억제된 장기를 생산할 수 있다.