Development and Application of a Semi quantitative Scoring Method for Ultrastructural Assessment of Acute Stress in Pancreatic Islets.
Development and Application of a Semi quantitative Scoring Method for Ultrastructural Assessment of Acute Stress in Pancreatic Islets.
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DOI：
10.1097/txd.0000000000001271
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发表时间：
2022-01
影响因子：
2.3
通讯作者：
Scott WE 3rd
中科院分区：
文献类型：
--
作者：
Dyson NJ;Kattner N;Honkanen-Scott M;Hunter B;Doyle JA;White K;Davey TS;Ploeg RJ;Bury YA;Tiniakos DG;Shaw JAM;Scott WE 3rd
胰腺和胰岛移植结果受到供体死亡、器官获取、处理和移植过程中急性应激对内分泌细胞损伤的负面影响。在这里，我们报告了一种新的电子显微镜评分系统，纽卡斯尔胰腺内分泌应激评分（NPESS）。NPESS是从我们先前验证的胰腺外分泌腺泡细胞评分方法中改编和扩展的，产生了4分制（0-3），将内分泌细胞核、线粒体、内质网、细胞质空泡化和分泌颗粒耗竭的超微结构病理分类，最大加性评分为15。我们将NPESS应用于脑干（DBD）和循环（DCD）死亡后死亡器官供体队列，冷缺血时间范围广泛（3.6-35.9 h），包括3名1型和3名2型糖尿病供体，以评估原位胰岛（n = 30）以及胰腺（n = 3）分离前和分离后。在DBD胰腺中，NPESS与冷缺血时间（头部：r = 0.55; P = 0.02）和镜像外分泌评分（r = 0.48; P = 0.01）相关。当按内分泌表型分层时，具有异质形态颗粒的细胞的评分高于α、β和δ细胞（P < 0.0001）。观察到内分泌-外分泌混合形态的细胞与NPESS增加相关（P = 0.02）。胰岛分离与NPESS改善相关（原位：8.39 ± 0.77 [平均值± SD];分离后：5.44 ± 0.31; P = 0.04）。NPESS提供了一个强大的方法，半定量评分的亚细胞超微结构的变化，在人胰腺内分泌细胞原位和胰岛分离与实用程序的无偏评价急性应激器官移植研究。
Pancreas and islet transplantation outcomes are negatively impacted by injury to the endocrine cells from acute stress during donor death, organ procurement, processing, and transplant procedures. Here, we report a novel electron microscopy scoring system, the Newcastle Pancreas Endocrine Stress Score (NPESS). NPESS was adapted and expanded from our previously validated method for scoring pancreatic exocrine acinar cells, yielding a 4-point scale (0–3) classifying ultrastructural pathology in endocrine cell nuclei, mitochondria, endoplasmic reticulum, cytoplasmic vacuolization, and secretory granule depletion, with a maximum additive score of 15. We applied NPESS in a cohort of deceased organ donors after brainstem (DBD) and circulatory (DCD) death with a wide range of cold ischemic times (3.6–35.9 h) including 3 donors with type 1 and 3 with type 2 diabetes to assess islets in situ (n = 30) in addition to pancreata (n = 3) pre- and postislet isolation. In DBD pancreata, NPESS correlated with cold ischemic time (head: r = 0.55; P = 0.02) and mirrored exocrine score (r = 0.48; P = 0.01). When stratified by endocrine phenotype, cells with granules of heterogeneous morphology had higher scores than α, β, and δ cells (P < 0.0001). Cells of mixed endocrine-exocrine morphology were observed in association with increased NPESS (P = 0.02). Islet isolation was associated with improved NPESS (in situ: 8.39 ± 0.77 [Mean ± SD]; postisolation: 5.44 ± 0.31; P = 0.04). NPESS provides a robust method for semiquantitative scoring of subcellular ultrastructural changes in human pancreatic endocrine cells in situ and following islet isolation with utility for unbiased evaluation of acute stress in organ transplantation research.