Preliminary report of perioperative monitoring of six-gene-edited pig-to-cynomolgus monkey kidney xenotransplantation
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摘要:
目的 探讨六基因编辑猪-非人灵长类动物异种肾移植模型的构建。 方法 将人源化基因编辑猪（GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM）的肾脏移植给食蟹猴，观察受体存活情况及恢复血流灌注后肾脏情况。定期监测肾脏实质回声、血流变化及大小；进行血常规、肾功能检测及电解质检测；监测尿液、粪便及体质量动态变化。食蟹猴生命终点取移植肾、心脏、肝脏、脾脏、肺脏、盲肠行病理学检查。 结果 受体术后7 d死亡。恢复血流后，肾脏灌注良好，器官质地柔软，色泽正常。受体生命终点时见肾脏腹侧有少量脓性分泌物附着，明显充血肿大，呈“大红肾”外观。术后随时间延长，肾实质回声增高，血流减少，皮质逐渐增厚，肾周及腹腔有少量积液。受体术后外周血红细胞、血红蛋白、白蛋白及血小板进行性下降，血清肌酐在术后7 d升高至308 μmol/L，K+变化不大。术后即有淡黄色尿液排出，术后3 h内即恢复饮食及饮水并排淡黄色成形大便1次。术后1 d内尿液颜色逐渐变淡红至恢复正常，尿常规检测结果相符。术后2 d晨起排褐色血便2次，量较多，予以奥美拉唑进行抑酸治疗，至术后4 d大便恢复正常。β2-微球蛋白在术后7 d增至0.75 mg/L。体质量增加1.7 kg。尸检病理学检查发现移植肾间质水肿出血，大量淋巴细胞和巨噬细胞浸润，小动脉壁及动脉腔内淋巴细胞浸润，有动脉炎改变；盲肠间质有淋巴细胞浸润；脾脏组织有淤血表现；其余器官未见明显异常改变。 结论 人源化基因编辑猪-非人灵长类动物异种肾移植模型构建获得初步成功，术后受体存活时间达1周。 -
关键词:
- 异种移植 /
- 肾移植 /
- 基因编辑猪 /
- 食蟹猴 /
- 排斥反应 /
- α-1, 3-半乳糖苷转移酶 /
- β-1, 4-N-乙酰氨基半乳糖转移酶2 /
- 补体调节蛋白
Abstract:Objective To investigate the establishment of a six-gene-edited pig-to-non-human primate kidney xenotransplantation model. Methods The kidney of humanized genetically-edited pig (GTKO/β4GalNT2KO/CMAHKO/hCD55/hCD46/hTBM) was transplanted into a cynomolgus monkey. The survival of the recipient and kidney condition after blood perfusion were observed. The parenchymal echo, blood flow changes, and size of the kidney were monitored on a regular basis. Routine blood test, kidney function test and electrolyte assessment were carried out. Dynamic changes of urine, feces and body mass were monitored. At the end of life, the transplant kidney, heart, liver, spleen, lung, and cecum were collected for pathological examination. Results The recipient died at postoperative 7 d. After blood flow was restored, the kidney was properly perfused, the organ was soft and the color was normal. At the end of the recipient's life, a slight amount of purulent secretion was attached to the ventral side of the kidney, with evident congestion and swelling, showing the appearance of "red kidney". Postoperatively, the echo of renal parenchyma was increased, blood flow was decreased, the cortex was gradually thickened, and a slight amount of effusion surrounded the kidney and abdominal cavity over time. In the recipient, the amount of peripheral red blood cells, hemoglobin, albumin, and platelets was progressively decreased, and serum creatinine level was increased to 308 μmol/L at postoperative 7 d, whereas the K+ concentration did not significantly change. Light yellow urine was discharged immediately after surgery, diet and drinking water were resumed within postoperative 3 h, and light yellow and normal-shape stool was discharged. The reddish urine was gradually restored to normal color within postoperative 1 d, which were consistent with the results of the routine urine test. A large amount of brown bloody stool was discharged twice in the morning of 2 d after surgery. Omeprazole was given for acid suppression, and the stool returned to normal at postoperative 4 d. The β2-microglobulin level was increased to 0.75 mg/L at postoperative 7 d. The body mass was increased by 1.7 kg. Autopsy pathological examination showed interstitial edema and bleeding of the transplant kidney, a large amount of infiltration of lymphocytes and macrophages, infiltration of lymphocytes in the arteriole wall and arterial cavity, accompanied by arteritis changes, lymphocyte infiltration in the cecal stroma and congestion in the spleen tissues. No significant abnormal changes were observed in other organs. Conclusions The humanized genetically-edited pig-to-non-human primate kidney xenotransplantation model is successfully established, and postoperative survival of the recipient is 1 week. -
图 1 移植肾的动态变化过程
注：A图为受体食蟹猴术前状态；B图为供体基因编辑猪术前状态；C图为供肾切取前状态；D图为异种肾移植术后恢复血流灌注后；E图为尸检肾脏外观；F图为解剖后肾脏状况。
Figure 1. The dynamic changes course of the transplant kidney
图 2 肾脏超声检查结果
注：A、D图为术前肾脏状况；B、E图为术后3 d肾脏状况；C、F图为术后7 d肾脏状况。
Figure 2. Ultrasound examination results of kidney
图 3 受体移植前后检验指标动态变化过程
Figure 3. The dynamics changes of experiment parameters of the recipient before and after transplantation
图 4 受体术后每日尿液指标及体质量动态变化
Figure 4. Dynamic changes of daily urine and body weight of the recipient after surgery
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