1、異種肝移植的研究進展及其所面臨的挑戰(zhàn)23目 錄 ONTENTS背景介紹GTKO豬為供體的相關(guān)研究術(shù)后凝血功能障礙相關(guān)研究異位肝移植1C2總結(jié)與展望5431背景介紹4異種移植（Xenotransplantation）不同種屬個體遺傳背景差異大嚴(yán)重的排斥反應(yīng)5器官移植需求表1 2016年美國器官移植等待名單表2 美國肝臟移植統(tǒng)計UNOS. United Network for Organ Sharing. Http:/. Updated 2016 Accessed December 21, 2016.6臨床病例1、美國洛杉磯西代細納醫(yī)院1992年野生型大白豬的肝臟膽汁流出存活24h2、美國Mako

2、wka等1995年野生型大白豬的肝臟血漿洗脫法和特異性抗體移出法肝存活20h，受體存活34h抗體介導(dǎo)的超急性排斥反應(yīng)Makowa, L, et al. The use of a pig liver xenograft for temporary support of a patient with fulminant hepatic failure. Transplantation59.12(1995):1654-9.72GTKO豬為供體的相關(guān)研究8GTKO豬靈長類動物體內(nèi)的抗-1,3-半乳糖的異種反應(yīng)性天然抗體豬血管內(nèi)皮細胞上的-1,3-半乳糖抗原-1,3-半乳糖苷轉(zhuǎn)移酶（-1,3-galac

3、tosy transferase, -GT）基因敲除小型豬（GTKO豬）Ekser, B, et al. Genetically-engineered pig-to-baboon liver xenotransplantation: histopathology of xenografts and native organs. Plos One 7.1(2011): e29720.9Figure 1. Time-line in experimental pig liver xenotransplantation in NHPs. a-CD154 = anti-CD154 monoclonal

4、antibodies; ALG,antilymphocyte globulin; ATG, antithymocyte globulin; Aza, azathioprine; Bela, belatacept; BM, bone marrow; Cs, corticosteroids; CsA, cyclosporine;CVF, cobra venom factor; CyP, cyclophosphamide; FK, tacrolimus; Gal abs, extracorporeal anti-Gal antibody adsorption; GTKO, 1,3-galactosy

5、ltransferase gene-knockout; hCD46, expression of the human regulator of complement, hCD46; hCD55, expression of the humanregulator of complement, CD55; HLT, heterotopic liver transplantation; HT, H-transferase; LoCD2b, anti-CD2 monoclonal antibody; MMF,mycophenolate mofetil; OLT, orthotopic liver tr

6、ansplantation; WBI, whole body irradiation; WT, wild-type.Cooper, D. K., et al. Pig Liver Xenotransplantation: A Review of Progress Toward the Clinic. Transplantation 100(2016).10GTKO/hCD46豬Figure 2 (A) Platelet counts after GTKO/hCD46 orthotopic pig liver transplantation in baboons (n = 6) that sur

7、vived from 4 to 7 days. (B) Histopathology of a GTKO/hCD46 orthotopic pig liver graft in a baboon that survived for 6 daysEkser, B, et al. Impact of thrombocytopenia on survival of baboons with genetically modified pig liver transplants: clinical relevance. American Journal of Transplantation 10.2(2

8、010):273-85.11GTKO豬與-氨基乙酸Figure 3. Thrombocytes were lost immediately after reperfusion.Figure 4. Normal liver tissue without any evidence of rejection,haematoxylin and eosin.Kim, K, et al. Up to 9-day survival and control of thrombocytopenia following alpha1,3-galactosyl transferase knockout swine

9、liver xenotransplantation in baboons. Xenotransplantation 19.4(2012):256-64.123術(shù)后凝血功能障礙相關(guān)研究13供體內(nèi)皮細胞Burcin Ekser , et al. Potential factors influencing the development of thrombocytopenia and consumptive coagulopathy after genetically modified pig liver xenotransplantation. Transplant International 2

10、5.8(2012):882-96.Figure 5 The expression of CD106 of genetically-engineered pig liver grafts were examined pre-TX, 2h after reperfusion, and at the time of death or euthanasia (days 47). 14受體組織因子的激活Figure 6 TF immunohistochemistry of human and pig TF in donor and recipient livers before, 2 hours aft

11、er, and 48 hours after transplantation.Figure 7 Measurement of endogenous tissue factor activity for recombinant human TFPI, recombinant pig TFPI, and no TFPI control.Ji, H., et al. Pig BMSCs Transfected with Human TFPI Combat Species Incompatibility and Regulate the Human TF Pathway in Vitro and in

12、 a Rodent Model. Cellular Physiology & Biochemistry International Journal of Experimental Cellular Physiology Biochemistry & Pharmacology 36.1(2015):233-49.15SIRP-/CD47 Navarroalvarez, Nalu, and Y. G. Yang. CD47: a new player in phagocytosis and xenograft rejection. Cellular & Molecular Immunology 8

13、.4(2011):285-8.Figure 8(A) SIRP-CD47 corss-species incompatibility.16SIRP-/CD47 Navarroalvarez, Nalu, and Y. G. Yang. CD47: a new player in phagocytosis and xenograft rejection. Cellular & Molecular Immunology 8.4(2011):285-8.Figure 8(B) SIRP-CD47 corss-species incompatibility.17去唾液酸糖蛋白1(Asialoglyco

14、protein receptor 1,ASGR 1)Ahrens, H. E., et al. siRNA mediated knockdown of tissue factor expression in pigs for xenotransplantation. American Journal of Transplantation 15.5(2015):14071414.Figure 9 Potential mechanism of human platelet phagocytosis by pig livers18GGTA1/CMAH基因Butler, J. R., et al. S

15、ilencing Porcine CMAH and GGTA1 Genes Significantly Reduces Xenogeneic Consumption of Human Platelets by Porcine Livers. Transplantation 100.3(2016).Figure 10 A continuous-pass model for the xenogeneic consumption of human platelets by modified porcine livers.194異位肝移植20異位肝移植Cooper, D. K., et al. Pig

16、 Liver Xenotransplantation: A Review of Progress Toward the Clinic. Transplantation 100(2016).Figure 11 Surgical technique of pig left liver lobe transplantation in Tibetanmonkeys.21異位肝移植Cooper, D. K., et al. Pig Liver Xenotransplantation: A Review of Progress Toward the Clinic. Transplantation 100(

17、2016).原位肝移植術(shù)切除受體肝臟后原位植入豬肝異位肝移植術(shù)優(yōu)點：不切除受體原肝保留原肝合成凝血因子和抗凝蛋白的功能切除受體脾臟控制術(shù)后免疫排異反應(yīng)224總結(jié)與展望23 總結(jié)與展望Yang, L., et al. Genome-wide inactivation of porcine endogenous retroviruses (PERVs). Science 350.6264(2015).倫理替代輔助供體 新型轉(zhuǎn)基因豬、嵌合器官供體受體 新型抗凝血、抗排異藥物生物安全性 豬逆轉(zhuǎn)錄病毒（Porcine endogenous retrovirus, PERV）參考文獻1 UNOS. Uni

18、ted Network for Organ Sharing. Http:/. Updated 2016 Accessed December 21, 2016.2 Makowa L, Cramer D V, Hoffman A, et al. The use of a pig liver xenograft for temporary support of a patient with fulminant hepatic failure.J. Transplantation, 1995, 59(12):1654-9.3 Ekser B, Klein E, He J, et al. Genetic

19、ally-engineered pig-to-baboon liver xenotransplantation: histopathology of xenografts and native organs.J. Plos One, 2011, 7(1): e29720. 4 Cooper D K, Dou D F, Tao K S, et al. Pig Liver Xenotransplantation: A Review of Progress Toward the Clinic.J. Transplantation, 2016, 100.5 Ekser B, Long C, Echev

20、erri G J, et al. Impact of thrombocytopenia on survival of baboons with genetically modified pig liver transplants: clinical relevance.J. American Journal of Transplantation, 2010, 10(2):273-85.6 Kim K, Schuetz C, Elias N, et al. Up to 9-day survival and control of thrombocytopenia following alpha1,

21、3-galactosyl transferase knockout swine liver xenotransplantation in baboons.J. Xenotransplantation, 2012, 19(4):256-64.7 Burcin Ekser, Chih C. Lin, Long C, et al. Potential factors influencing the development of thrombocytopenia and consumptive coagulopathy after genetically modified pig liver xeno

22、transplantationJ. Transplant International, 2012, 25(8):882-96.8 Ji H, Li X, Yue S, et al. Pig BMSCs Transfected with Human TFPI Combat Species Incompatibility and Regulate the Human TF Pathway in Vitro and in a Rodent Model.J. Cellular Physiology & Biochemistry International Journal of Experimental Cellular Physiology Biochemistry &