Molecular Profiling of Pancreatic Pathophysiology by Imaging Mass Spectrometry

通过成像质谱法对胰腺病理生理学进行分子分析

• 批准号: 8908974
• 负责人: Boone M. Prentice
• 金额: $ 5.24万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-22 至 2016-07-21
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8908974
• 关键词: Adolescent; Adult; Affect; Alpha Cell; Amputation; Anatomy; Antibodies; Arachidonic Acids; Architecture; Beta Cell; Biochemical; Biological; Biological Markers; Biology; Blindness; Cell Death; Cells; Cellular Structures; Ceramides; Cessation of life; Chemistry; Collaborations; Complex; Development; Diabetes Mellitus; Diagnosis; Disease; Disease Progression; Endocrine; Environment; Functional disorder; Future; Goals; Health; Heart Diseases; Homeostasis; Hormones; Human; Hypertension; Image; Imaging technology; Immunohistochemistry; In Situ; Insulin Resistance; Interdisciplinary Study; Islet Cell; Islets of Langerhans; Kidney Diseases; Knowledge; Leptin; Lipids; Location; Longitudinal Studies; Maps; Mass Spectrum Analysis; Measurement; Measures; Medical; Metabolic; Metabolism; Methods; Molecular; Molecular Profiling; Morphology; Mus; Nature; Neuropathy; Non-Insulin-Dependent Diabetes Mellitus; Obesity; Organ; Organogenesis; Pancreas; Pathway interactions; Phospholipids; Play; Population; Process; Productivity; Proteins; Research; Resolution; Role; Scientist; Signal Pathway; Signal Transduction; Signaling Molecule; Specimen; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Staging; Stroke; Sulfoglycosphingolipids; Techniques; Technology; Testing; Tissues; Training; Translating; United States; base; blood glucose regulation; career; cell type; cost; design; diabetic; endocrine pancreas development; glucose transport; insight; interdisciplinary approach; interest; islet; lipid metabolism; loss of function; molecular array; molecular imaging; mouse model; novel; novel strategies; outcome forecast; post-doctoral training; public health relevance; therapeutic target

 DESCRIPTION (provided by applicant): While hormone-secreting pancreatic islet cells (e.g.,  cells and ß cells) have been identified to play key roles in glucose homeostasis, our understanding of the processes important to normal pancreatic organogenesis and leading to altered metabolic states such as type 2 diabetes (T2D) is still limited. The low availability of human islets and the lack of experimental methods to comprehensively analyze islet cells and their tissue environments restrict our ability to map cell signaling pathways and metabolism. Despite substantial progress towards treatments and cures, much remains unknown about the specific signaling and damage pathways that contribute to ß-cell dysfunction. A more fundamental understanding of islet development and diabetes pathophysiology is required to help direct diagnoses, prognoses, treatments, and therapies and likely begins at the molecular level. In this way, novel biochemical understandings can be translated to insights on pancreatic development and disease progression, ultimately leading to the identification of new potential biomarkers and therapeutic targets. The proposed discovery-based research herein will provide molecular-level insight on pancreatic islet architecture, function, and development by combining a form of new molecular imaging technology, matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI IMS), with existing validated approaches. This technology is well suited to measuring the complex array of molecular information (e.g., lipids and proteins) present in cellular tissue and provides for an untargeted measurement method, which is ideal for the discovery of new molecules important to pancreatic function. We aim to utilize such in situ tissue-based IMS techniques to develop much more complete molecular maps for the different pancreatic endocrine cell types, allowing us to gain insight as to the cell signaling pathways involved in pancreatic development and disease progression. These IMS techniques will be used to specifically define the lipid and protein molecular signatures of normal mouse islets and correlate these maps to functional changes observed in the presence of obesity and insulin resistance. This will allow us to establish those lipid and protein signaling molecules crucial to lipotoxicity and islet dysfunction in the well-studied ob/ob mouse model. Finally, these IMS approaches will be expanded to perform a longitudinal study identifying differences in protein and lipid profiles of human pancreatic islet cells throughout organogenetic development from the juvenile to the adult stages, and determining how these biochemical changes relate to morphological and architectural changes in the tissue. The molecular profiles produced by these interdisciplinary studies will provide insights on the cell signaling mechanisms important in pancreatic maturation and disease progression and will aid in the design of novel therapies and strategies to sustain islet mass and function.

 描述（由申请人提供）：虽然已确定分泌激素的胰岛细胞（例如 α 细胞和 ß 细胞）在葡萄糖稳态中发挥关键作用，但我们对正常胰腺器官发生和导致代谢状态等重要过程的理解由于 2 型糖尿病 (T2D) 的数量仍然有限，而且缺乏全面分析胰岛细胞及其组织环境的实验方法限制了我们绘制图谱的能力。尽管在治疗和治愈方面取得了实质性进展，但对于导致 β 细胞功能障碍的特定信号传导和损伤途径仍知之甚少，需要对胰岛发育和糖尿病病理生理学有更基本的了解，以帮助指导诊断和预后。通过这种方式，新的生化理解可以转化为对胰腺发育和疾病进展的见解，最终导致新的潜在生物标志物和治疗靶点的识别。研究指南将通过将新型分子成像技术、基质辅助激光解吸/电离成像质谱（MALDI IMS）与现有经过验证的方法相结合，提供对胰岛结构、功能和发育的分子水平见解。测量细胞组织中存在的复杂分子信息（例如脂质和蛋白质），并提供一种非靶向测量方法，这对于发现对胰腺功能重要的新分子来说是理想的。我们的目标是利用这种原位方法。基于组织的 IMS 技术可以为不同的胰腺内分泌细胞类型开发更完整的分子图谱，使我们能够深入了解参与胰腺发育和疾病进展的细胞信号传导途径。这些 IMS 技术将用于具体定义脂质。正常小鼠胰岛的蛋白质分子特征，并将这些图谱与肥胖和胰岛素抵抗存在下观察到的功能变化相关联，这将使我们能够在经过充分研究的小鼠中建立对脂毒性和胰岛功能障碍至关重要的脂质和蛋白质信号分子。观察鼠标最后是这些模型。 IMS 方法将扩展到进行纵向研究，识别从幼年到成年阶段的整个器官发育过程中人类胰岛细胞的蛋白质和脂质谱的差异，并确定这些生化变化与组织的形态和分子结构变化之间的关系。这些跨学科研究产生的概况将提供对胰腺成熟和疾病进展中重要的细胞信号传导机制的见解，并将有助于设计维持胰岛质量和功能的新疗法和策略。