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: 10400943
• 负责人: MARK A. ATKINSON
• 金额: $ 41.44万
• 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 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10400943
• 关键词: 3-Dimensional; Acinar Cell; Acinus organ component; Address; Affect; 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 autoantibodies; Diabetes prevention; Digestion; Disease; 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 - Cell Movement; Stimulus; Structure of beta Cell of islet; Tail; Techniques; Technology; Testing; Three-dimensional analysis; Tissue Donors; Tissue Slice Technology; Tissues; Trypsinogen; Tumor-infiltrating immune cells; Variant; Vascularization; Weight; autoimmune pathogenesis; blood glucose regulation; cellular imaging; density; diabetes pathogenesis; experience; imaging study; 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赠者（NPOD）等计划，高价 现在可以使用具有各个阶段和持续时间的1型糖尿病（T1D）的人的组织。因此， 我们对T1D开发基础的致病事件的集体知识已大大提高。 但是，仅评估人胰腺功能和形态特征的研究尚未 进行了执行，限制了我们对健康中控制器官生理的过程的理解 T1D中的病理生理学。尽管有重要的研究资源，但对孤立胰岛的调查是 被隔离程序混淆，隔离程序引起了炎症反应，形态修饰， 并改变基因表达。此外，隔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，胰岛与他们的分离 周围的组织消除了有关局部相邻元素的任何影响和信息， 很重要，尤其是在病理环境中。我们相信胰岛的特征和外分泌 组织（功能，调节和细胞相互作用）及其在T1D发病机理中的作用 通过研究含有完整小岛和acini的可行人类胰组织的研究。我们建议 利用非常新颖的胰腺组织切片技术来大大扩展我们对内分泌的知识 以及通过正常（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。该技术最初由Spier博士（MPD/PI，HMGU）建立，生产的“切片” 新鲜的胰组织，具有最小的机械损伤，没有酶消化；原位启用 在相对不受干扰的组织设置中研究胰岛细胞生物学。以下功能模拟 内分泌（胰岛素和胰高血糖素）和外分泌分泌（淀粉酶，脂肪酶，胰蛋白酶原）的测定法以及 细胞信号传导（CA2+通量）的动态成像研究，将通过3D形态计量法固定并分析切片。 在将要解决的许多问题中，我们试图提高对功能的理解 小叶和区域（即头部，身体或尾巴）异质胰形态的含义，包括 胰岛密度，大小和细胞组成，外分泌酶的表达和分布，以及血管和血管和 神经元密度。此外，我们旨在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/器官质量的丧失）。所以，我们 姿势测试以下假设：胰岛炎的分散器官体重减轻以及小叶异质性 人类T1D胰腺内的β细胞破坏是区域间和骨内的结果 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 和外分泌生理学和病理生理学到其在胰腺内的位置和细胞特征 周围的微环境；应影响预防/逆转的尝试的努力。