The science and engineering of stem cell-derived organoids-examples from hepatic,biliary, and pancreatic tissues

The field of organoid engineering promises to revolutionize medicine with wide-ranging applications of scientific, engineering, and clinical interest, including precision and personalized medicine, gene editing, drug development, disease modelling, cellular therapy, and human development. Organoids are a three-dimensional (3D) miniature representation of a target organ, are initiated with stem/progenitor cells, and are extremely promising tools with which to model organ function. The biological basis for organoids is that they foster stem cell self-renewal, differentiation, and self-organization, recapitulating 3D tissue structure or function better than two-dimensional (2D) systems. In this review, we first discuss the importance of epithelial organs and the general properties of epithelial cells to provide a context and rationale for organoids of the liver, pancreas, and gall bladder. Next, we develop a general framework to understand self-organization, tissue hierarchy, and organoid cultivation. For each of these areas, we provide a historical context, and review a wide range of both biological and mathematical perspectives that enhance understanding of organoids. Next, we review existing techniques and progress in hepatobiliary and pancreatic organoid engineering. To do this, we review organoids from primary tissues, cell lines, and stem cells, and introduce engineering studies when applicable. We discuss non-invasive assessment of organoids, which can reveal the underlying biological mechanisms and enable improved assays for growth, metabolism, and function. Applications of organoids in cell therapy are also discussed. Taken together, we establish a broad scientific foundation for organoids and provide an in-depth review of hepatic, biliary and pancreatic organoids.     

超声对原发性胆汁性肝硬化合并自身免疫性甲状腺炎的诊断价值和相关性分析

摘要 目的：探讨超声对原发性胆汁性肝硬化合并自身免疫性甲状腺炎的诊断价值以及相关性。方法：选取2017年1月~2019年12月我院收治的90例原发性胆汁性肝硬化合并自身免疫性甲状腺炎患者为研究组,根据患者病情的不同将其分为早期组（n=48）和晚期组（n=42）。另选取50例健康志愿者为对照组,对所有患者进行超声诊断,比较两组患者的门静脉血流参数、肝动脉血流参数、肝中静脉血流参数以及脾静脉血流参数。结果：晚期组的门静脉主干内径（portal vein diameter,PVD）明显高于早期组和对照组（P<0.05）,晚期组的平均流速（portal vein velocity,PVV）明显低于早期组和对照组（P<0.05）,三组患者的门静脉血流量（perceptual video quality,PVQ）差异无统计学意义（P>0.05）。晚期组的肝动脉内径（hepatic artery inner diameter,HAD）、阻力指数（resistance index,RI）、收缩期峰值流速（peak systolic velocity/end diastolic velocity,HAVmax）、血流量（blood flow,HAQ）均明显大于对照组（P<0.05）。晚期组的肝中静脉内径（middle hepatic vein diameter,MHAD）、最高平均流速（maximum average flow rate ,MHV Vm）、最大血流量（maximum blood flow,MHVQ）均较早期组和对照组减少（P<0.05）。晚期组的脾静脉内径（splenic vein diameter,SVD）明显高于对照组和早期组（P<0.05）,晚期组的脾静脉平均流速（splenic vein mean flow rate,SVm）、脾静脉血流量 (splenicveinflow, SVF)与对照组、早期组相比差异无统计学意义（P>0.05）;经过相关性分析肝脏超声指标与甲状腺超声指标均呈现正相关关系（P＜0.05）。结论：超声在原发性胆汁性肝硬化合并自身免疫性甲状腺炎患者中应用具有很高的价值,通过对原发性胆汁性肝硬化超声指标推测自身免疫性甲状腺炎病情程度,患者的超声图像具有一定的特征性,可以用以患者的诊断和病因分析,值得在临床中应用推广。     

Hepatic Deletion of X-Box Binding Protein 1 in FXR Null Mice Leads to Enhanced Liver Injury

FXR regulates bile acid metabolism, and FXR null (Fxr^?/?) mice have elevated bile acid levels and progressive liver injury. The inositol-requiring enzyme 1α/X-box binding protein 1 (XBP1) pathway is a protective unfolded protein response pathway activated in response to endoplasmic reticulum stress. Here, we sought to determine the role of the inositol-requiring enzyme 1α/XBP1 pathway in hepatic bile acid toxicity using the Fxr^?/? mouse model. Western blotting and quantitative PCR analysis demonstrated that hepatic XBP1 and other unfolded protein response pathways were activated in 24-week-old Fxr^?/? compared with 10-week-old Fxr^?/? mice but not in WT mice. To further determine the role of the liver XBP1 activation in older Fxr^?/? mice, we generated mice with whole-body FXR and liver-specific XBP1 double KO (DKO, Fxr^?/?Xbp1^LKO) and Fxr^?/?Xbp1^fl/fl single KO (SKO) mice and characterized the role of hepatic XBP1 in cholestatic liver injury. Histologic staining demonstrated increased liver injury and fibrosis in DKO compared with SKO mice. RNA sequencing revealed increased gene expression in apoptosis, inflammation, and cell proliferation pathways in DKO mice. The proapoptotic C/EBP-homologous protein pathway and cell cycle marker cyclin D1 were also activated in DKO mice. Furthermore, we found that total hepatic bile acid levels were similar between the two genotypes. At age 60 weeks, all DKO mice and no SKO mice spontaneously developed liver tumors. In conclusion, the hepatic XBP1 pathway is activated in older Fxr^?/? mice and has a protective role. The potential interaction between XBP1 and FXR signaling may be important in modulating the hepatocellular cholestatic stress responses.     

Liver-specific T regulatory type-1 cells program local neutrophils to suppress hepatic autoimmunity via CRAMP

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Human biliary epithelial cells from discarded donor livers rescue bile duct structure and function in a mouse model of biliary disease

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脐带间充质干细胞治疗熊去氧胆酸应答不佳的原发性胆汁性胆管炎的临床观察

目的探讨脐带间充质干细胞（UC-MSCs）输注治疗熊去氧胆酸（UDCA）应答不佳的原发性胆汁性胆管炎（PBC）患者的安全性和有效性，分析影响UC-MSCs疗效应答的相关因素。 方法选取解放军总医院第五医学中心2010年8月至2017年10月接受UC-MSCs输注治疗UDCA应答不佳的PBC患者29例。患者均以4周为间隔给予3次外周静脉输注细胞1.0×10⁶个/kg。通过实验室指标、生命体征及不良事件发生情况评估UC-MSCs治疗的安全性。通过患者临床症状、肝功指标和Child-Pugh评分评估治疗的有效性。以"巴黎Ⅰ标准"作为疗效标准，评价患者UC-MSCs治疗后的疗效应答情况，比较有效患者及无效患者基线临床症状和肝脏功能差异，分析影响UC-MSCs疗效的相关因素。采用独立样本t检验分析年龄；采用Mann-Whitney U检验比较两组UDCA治疗时间、激素治疗时间、实验室数据等，采用Wilcoxon秩和检验比较组间数据；采用χ²检验比较性别、临床症状和Child-Pugh分级等指标。多因素Cox回归分析对影响UC-MSCs疗效的相关因素。 结果1例患者在治疗后因合并严重感染出现高热，所有患者未出现UC-MSCs相关严重不良事件。UC-MSCs输注后与基线相比，患者的血清碱性磷酸酶（ALP）[281.00 （182.50，428.50）比201.00 （149.50，402.00）]、γ-谷氨酰转移酶（GGT）[156.00 （73.00，390.00）比84.00 （43.50，312.50）]、总胆固醇（TC）[5.10 （3.14，7.69）比3.94 （3.00，6.01）]均下降（P < 0.05）。其中，9例（31﹪）患者治疗后疗效明显，达到"巴黎Ⅰ标准"，与无效组患者基线相比，有效组患者天冬氨酸转氨酶（AST）[93.50 （77.75，100.75）比53.00 （46.00，78.00）]、ALP[342.00 （237.25，516.00）比185.00 （152.50，295.50）]、总胆红素（TBIL）[58.50 （33.45，69.33）比13.10 （11.25，20.25）]均下降（P < 0.05）。多因素Cox回归分析显示，血清TBIL是影响UC-MSCs疗效的重要独立因素[HR为0.817 （95﹪CI：0.715 ~ 0.935），P < 0.05]。 结论UC-MSCs输注对UDCA治疗应答不佳的PBC患者是安全可行的且耐受性良好，部分患者肝功能得到一定的改善。血清TBIL是影响UC-MSCs治疗疗效的独立重要因素，提示在疾病进展早期TBIL较低的阶段进行UC-MSCs治疗可能有效改善和减缓患者疾病进程。