Regional and lobular heterogeneity of human pancreas morphology and function in type 1 diabetes pathogenesis

1型糖尿病发病机制中人胰腺形态和功能的区域和小叶异质性

• 批准号: 10617206
• 负责人: MARK A. ATKINSON
• 金额: $ 40.7万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-24 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10617206
• 关键词: 3-Dimensional; Acinar Cell; Acinus organ component; Address; Affect; Age; Amylases; Area; Atrophic; Autoantibodies; Autoimmunity; Beta Cell; Biological Assay; Blood Vessels; Body Weight decreased; Body mass index; C-Peptide; Cell Communication; Cell physiology; Cells; Cellular biology; Cessation of life; Characteristics; Data; Development; Diabetes Mellitus; Diabetes prevention; Digestion; Disease; Disparate; Disparity; Elements; Endocrine; Environment; Enzymes; Event; Exocrine pancreas; Extracellular Matrix; Functional disorder; Gene Expression; Glucagon; Head; Health; Heterogeneity; Histologic; Hormones; Human; Immune; In Situ; Individual; Infiltration; Inflammatory Response; Insulin; Insulin-Dependent Diabetes Mellitus; Investigation; Islet Cell; Islets of Langerhans; Knowledge; Lipase; Lobular; Lobule; Location; Measures; Mechanics; Mediating; Modification; Molecular; Morphology; Mus; Neurons; Organ; Organ Donor; Organ Weight; Output; Pancreas; Pathogenesis; Pathogenicity; Pathologic; Pattern; Persons; Physiology; Positioning Attribute; Predisposition; Procedures; Process; Regulation; Resistance; Resources; Role; Serum; Signal Transduction; Slice; Sorting; Stimulus; Tail; Techniques; Technology; Testing; Tissue Donors; Tissue Slice Technology; Tissues; Trypsinogen; Variant; Vascularization; Weight; autoimmune pathogenesis; blood glucose regulation; cellular imaging; density; diabetes pathogenesis; experience; imaging study; immune cell infiltrate; improved; insulitis; islet; lymphatic vessel; morphometry; nerve supply; new therapeutic target; non-diabetic; novel; peripheral blood; preservation; prevent; programs; regional difference; response; sex; trait; two-dimensional

Through programs such as the Network for Pancreatic Organ donors with Diabetes (nPOD), highly valuable tissues from persons with various stages and durations of type 1 diabetes (T1D) are now available. As a result, our collective knowledge of the pathogenic events underlying T1D development has improved substantially. However, studies simultaneously assessing functional and morphological traits of the human pancreas have not been performed, limiting our understanding of the processes governing organ physiology in health versus pathophysiology in T1D. Despite being in important research resource, investigations of isolated islets are confounded by the isolation procedure, which induces an inflammatory response, morphological modifications, and altered gene expression. Furthermore, the isolation procedure relies on the structural integrity of the islets to withstand the enzymatic and mechanical sorting process. Most importantly, separation of islets from their surrounding tissue removes any influence and information regarding the local adjacent elements, which could be of importance, especially in pathological settings. We believe the characterization of islets as well as exocrine tissues (function, regulation, and cellular interactions) and their roles in the pathogenesis of T1D would be improved through studies of viable human pancreatic tissue containing intact islets and acini. We propose to utilize the extremely novel pancreas tissue slice technology to substantially expand our knowledge of endocrine and exocrine function and their interaction in normal (i.e., control) and T1D pancreas tissues obtained through the nPOD program. This technology, originally established by Dr. Speier (mPD/PI, HMGU), produces “slices” of fresh pancreatic tissue with minimal mechanical damage and without enzymatic digestion; enabling in situ investigations of islet cell biology in a relatively unperturbed tissue setting. Following functional stimulation assays for endocrine (insulin and glucagon) and exocrine secretion (amylase, lipase, trypsinogen) as well as dynamic imaging studies of cellular signaling (Ca2+ flux), slices will be fixed and analyzed by 3D morphometry. Amongst the many questions that will be addressed, we seek to improve our understanding on the functional implications of the lobular and regional (i.e., head, body or tail) heterogeneous pancreatic morphology, including islet density, size and cellular composition, exocrine enzyme expression and distribution, as well as vascular and neuronal density. Furthermore, we aim to address its role in the distinct lobular and regional pathological progression of T1D (i.e., insulitis, β-cell dysfunction and destruction, loss of acinar cell/organ mass). Thus, we pose to test the hypothesis that the disparate organ weight loss as well as the lobular heterogeneity of insulitis and β-cell destruction within the human T1D pancreas is the result of inter-regional and intra-lobular differences in endocrine and exocrine pancreas morphology and function. Our objective is to correlate islet and exocrine physiology and pathophysiology to their location within the pancreas and to cellular characteristics of the surrounding microenvironment; efforts that should impact attempts at T1D prevention/reversal.

通过诸如糖尿病胰腺器官捐献者网络 (nPOD) 等计划，非常有价值 因此，来自不同阶段和持续时间的 1 型糖尿病 (T1D) 患者的组织现已可用。 我们对 T1D 发展背后的致病事件的集体了解已大大提高。 然而，同时评估人类胰腺功能和形态特征的研究尚未 已进行，限制了我们对健康器官生理学控制过程的理解 T1D 的病理生理学 尽管正在进行重要的研究资源调查，但孤立的胰岛仍然存在。 被隔离程序所困扰，该程序会引起炎症反应、形态改变， 此外，分离过程依赖于胰岛的结构完整性。 承受酶促和机械分选过程，最重要的是，将胰岛与其分离。 周围组织消除了有关局部相邻元素的任何影响和信息，这可能 很重要，特别是在病理环境中。我们相信胰岛和外分泌的特征。 组织（功能、调节和细胞相互作用）及其在 T1D 发病机制中的作用将是 我们建议通过对含有完整胰岛和腺泡的活人胰腺组织的研究来改进。 利用极其新颖的胰腺组织切片技术大大扩展我们对内分泌的知识 正常（即对照）和 T1D 胰腺组织中的外分泌功能及其相互作用 nPOD 程序最初由 Speier 博士（mPD/PI，HMGU）建立，可生产“切片” 新鲜的胰腺组织，机械损伤最小，无需酶消化； 在功能刺激后相对不受干扰的组织环境中对胰岛细胞生物学进行研究。 内分泌（胰岛素和胰高血糖素）和外分泌（淀粉酶、脂肪酶、胰蛋白酶原）以及 细胞信号传导（Ca2+通量）的动态成像研究，切片将被固定并通过 3D 形态测量法进行分析。 在将要解决的许多问题中，我们寻求提高对功能性的理解 小叶和区域（即头部、身体或尾部）异质胰腺形态的影响，包括 胰岛密度、大小和细胞组成、外分泌酶表达和分布，以及血管和 此外，我们的目标是解决其在独特的小叶和区域病理中的作用。 T1D 的进展（即胰岛炎、β 细胞功能障碍和破坏、腺泡细胞/器官质量损失）。 检验以下假设：不同的器官重量减轻以及胰岛炎的小叶异质性 人类 T1D 胰腺内的 β 细胞破坏是区域间和小叶内的结果 内分泌和外分泌胰腺形态和功能的差异我们的目标是关联胰岛。 外分泌生理学和病理生理学及其在胰腺内的位置和细胞特征 周围微环境的影响；影响 T1D 预防/逆转尝试的努力。

项目成果

ACE2 chromogenic immunostaining protocol optimized for formalin-fixed paraffin-embedded human tissue sections.

针对福尔马林固定石蜡包埋的人体组织切片优化的 ​​ACE2 显色免疫染色方案。

• 发表时间: 2021
• 作者: Jorgensen, Marda;Joseph, Paul;Posgai, Amanda L;Vander Heide, Richard S;Kusmartseva, Irina;Atkinson, Mark A
• 通讯作者: Atkinson, Mark A