Analysis of molecular mechanism of immunological rejection of transplanted heart and development of gene therapy for heart rejection

同种异体心脏移植排斥反应仍然是心脏移植后的主要并发症。本项目旨在开发预防同种异体心脏移植急、慢性排斥反应的新型基因治疗方法。肿瘤坏死因子超家族成员LIGH结合疱疹病毒进入介质(HVEM)和淋巴毒素受体(LT_R)，在调节免疫应答中发挥重要作用。为了阐明移植物动脉疾病(GAD)的发病机制，我们研究了LIGH通路在GAD进展中的作用。我们在小鼠心脏移植模型中发现光通路不仅在调节T细胞的激活中起重要作用，而且在SMC增殖中也起着重要的调节作用。阻断光通路是预防广泛性阿尔茨海默病的有效途径。程序性死亡1(PD-1)是CD28家族的一员，参与免疫应答的负性调节，在外周免疫耐受的调节中起着特别重要的作用。程序性死亡--配体1…PD-L1在淋巴器官和非淋巴器官中的表达也较多。我们在小鼠心脏移植模型中研究了PD-1/PD-L1通路在急、慢性排斥反应中的作用。在完全同种异体模型中，给予抗PD-L1单抗可显著加速同种异体移植排斥反应。免疫组织化学显示浸润性细胞明显增多。在II类错配模型中，PD-L1在移植物冠状动脉内膜增厚的平滑肌细胞(SMC)中表达，抗PD-L1单抗的应用促进了GAD的进展。PD-1/PD-L1通路调节心脏移植的急性和慢性排斥反应。因此，对这一途径的控制应该为提高移植物存活率和抑制GAD提供一个有效的策略。肝细胞生长因子(HGF)是一种多效性因子，在组织修复和再生中发挥重要作用。我们展示了HGF在急性心脏排斥反应中的作用，并评估了它的免疫调节作用。肝细胞生长因子治疗显著延长了移植物存活时间，并诱导了40%的受者耐受。经HGF处理的同种异体移植物中，干扰素-α的表达降低，而白细胞介素10和转化生长因子-1的表达增强。提示HGF可改变辅助性T细胞介导的同种异体免疫反应。因此，HGF的应用可能成为预防心脏移植患者急性排斥反应的一种新的治疗方法。该项目的目标是开发基因治疗的临床应用，以最大限度地减少急性和慢性排斥反应。我们评估了核因子-kB诱饵在预防经皮冠状动脉介入治疗术后再狭窄中的作用。我们开始了临床试验。最初的病例是劳力性心绞痛，右冠状动脉中段(#2)狭窄。他接受了支架；我们在远端部位(#2)提供了核因子-kB诱饵，在近端部位(#1)没有诱饵。经皮冠状动脉介入治疗6个月后，与未使用诱骗基因相比，核因子-kB诱骗基因可抑制再狭窄。未观察到全身不良反应。这提示核因子-kB诱饵预防心脏病的临床安全性。因此，我们表明，这些方法可以应用于预防移植物衰竭，而不会产生全身不良反应。较少

Cardiac allograft rejection is still a major complication after heart transplantation. This project was aimed to develop novel gene therapies for preventing acute and chronic rejection of cardiac allograft. The TNF superfamily member LIGHT, which binds herpes virus entry mediator (HVEM) and lymphotoxin _receptor (LT_R), plays important roles in regulating the immune response. To clarify the mechanism underlying graft arterial disease (GAD), we investigated the role of the LIGHT pathway in the progression of GAD. We showed that the LIGHT pathway plays important roles in the regulation not only of T cell activation but also of SMC proliferation using mouse model of heart transplantation. Blockade of the LIGHT pathway is a promising avenue for the prevention of GAD. Programmed death 1 (PD-1), a member of the CD28 family, is involved in the negative regulation of the immune response and plays a particularly important role in the regulation of peripheral tolerance. Programmed death-ligand 1 … More (PD-L1) expression has been detected in lymphoid as well as non-lymphoid organs. We investigated the role of the PD-1/PD-L1 pathway in acute and chronic rejection in murine cardiac transplantation models. Administration of anti-PD-L1 mAb significantly accelerated allograft rejection in a fully allogeneic model. Immunohistochemistry showed significantly increased numbers of infiltrating cells. In a class II mismatch model, PD-L1 was expressed in smooth muscle cells (SMCs) of the thickened intima in the graft coronary artery, and the administration of anti-PD-L1 mAb enhanced the progression of GAD. The PD-1/PD-L1 pathway regulates both acute and chronic rejection in cardiac transplantation. Thus, control of this pathway should provide a potent strategy for improving graft survival and suppressing GAD. Hepatocyte growth factor (HGF) is a pleiotropic factor that plays an important role in tissue repair and regeneration. We show the effect of HGF on acute cardiac rejection and assess its immunomodulative role. HGF treatment resulted in a significant prolongation in graft survival, and tolerance was induced in 40% of the recipients. In HGF-treated allografts, the expression of interferon-_mRNA was decreased, and the expressions of interleukin-10 and transforming growth factor-_1 mRNA were enhanced. Our findings suggest that HGF could modify the alloimmuno response mediated by T helper cells. Thus, HGF administration could constitute a new therapeutic approach for prevention of acute rejection in patients undergoing cardiac transplantation. The goal of this project is to develop clinical application of gene therapy for minimizing acute and chronic rejection. We evaluated the utility of the NF-kB decoy for preventing restenosis after PCI. We started clinical trials. An initial case was suffering from effort angina with stenosis in the middle (#2) portions of the right coronary artery. He received stents ; we delivered the NF-kB decoy at the distal site (#2) and no decoy at the proximal site (#1). Six months after the PCI, NF-kB decoy suppressed the restenosis compared to no decoy transfection. No systemic adverse effects were observed. This suggests the clinical safety of NF-kB decoy for preventing heart disease. Thus, we showed that these methods could be applied to prevent graft failure without systemic adverse effects. Less