Induced Reactive Oxygen Species in a 3-D Human Pancreatitis Model in Vitro

体外 3D 人类胰腺炎模型中诱导的活性氧

基本信息

批准号：
8478094
负责人：
Gregory P Botta
金额：
$ 4.2万
依托单位：
DREXEL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8478094
关键词：
3-Dimensional Acinus organ component Acute Adhesions Adult Affect Alcoholic Pancreatitis Alcohols Antibodies Apoptosis Apoptotic Biological Models Cell Adhesion Molecules Cells Cessation of life Characteristics Clinical Confocal Microscopy Culture Media DNA Damage DNA Microarray Chip Desmoplastic Development Disintegrins Duct (organ) structure Ductal Ductal Epithelial Cell Ductal Epithelium Elasticity Environment Epithelial Epithelial Cells Ethanol Event Exhibits Exposure to Extracellular Matrix Future Generations Growth Factor Heavy Drinking Hemorrhagic Shock Human Hydrogels IL8 gene In Vitro Induction of Apoptosis Inflammation Mediators Inflammatory Injury Integrins Lipid Peroxidation Mammary gland Measures Microarray Analysis Modeling Morphogenesis Pain Pancreas Pancreatic duct Pancreatitis Pathogenesis Pathology Pathway interactions Physiological Production Reaction Reactive Oxygen Species Respiratory distress Rodent Role Signal Transduction Simulate Staining method Stains Testing Time Validation alcohol exposure alcohol response annexin A5 caspase-3 chemokine in vitro Model in vivo matrigel nestin protein neutralizing antibody novel polyacrylamide gels research study response scaffold stellate cell viscoelasticity

项目摘要

DESCRIPTION (provided by applicant): The most common cause of Pancreatitis is excessive alcohol intake which has the potential to induce reactive oxygen species (ROS) leading to DNA damage and apoptosis. In addition, characteristic pathological responses of pancreatitis, including inflammatory chemokines and a stiff, desmoplastic microenvironment may also contribute to this increased oxidative potential in pancreatic ductal cells. ROS can cause a severe inflammatory reaction instigating severe pain, hemorrhagic shock, acute respiratory distress, or death, however, no in vitro human model yet exists to understand this pathology. As such, we have begun to develop a novel, three-dimensional, human pancreatic duct model composed of nestin-positive, immortalized, pancreatic epithelial cells, Matrigel scaffolds, predetermined culture media, and specific growth factors. This model is characterized via confocal microscopy for cellular organization, polarization, and functionality similar to human mammary epithelial acini. Secondly, much progress has been made in understanding pancreatic stellate cell reactions to ethanol, but less is understood of the pancreatic epithelial cell response. Thus, we will analyze the effect of increasing concentrations of ethanol on ROS induction within these human pancreatic ducts via fluorescent staining and lipid peroxidation as well as the subsequent DNA damage response and apoptotic initiation events via the double strand break markers (H2AX and (SP1101 and Caspase-3, Bim, and Annexin V, respectively. Thirdly, pancreatic stellate cells have also been implicated in ROS production via inflammatory mediators, however, little has been advanced in the pancreatic epithelial cell response. As such, we will examine the direct initiation of ROS within the human pancreatic duct model via an inflammatory chemokine regularly found within the pancreatitis environment, IL-8. Through treatment with IL-8, we will determine the human pancreatic epithelial cell duct ROS-induced DNA damage and apoptosis response. Lastly, we will continue to verify the extent of ROS production within human pancreatic epithelial ducts by a stiffening extracellular matrix. By blocking pancreatic-specific integrin (3, (4 and (6 through antibodies/disintegrins and by DNA microarray validation of their expression, we will understand how exogenous elastic stiffness can affect ROS potential internal to pancreatic epithelial cells. We anticipate that the development of our in vitro, 3-D, human pancreatic epithelial duct model along with each of the three independent experiments will uncover parts of the etiologic pathway leading to pancreatitis in a human-relevant model and offer possible targets for future clinical therapy.

描述（由申请人提供）：胰腺炎的最常见原因是酒精摄入过多，有可能诱导活性氧（ROS）导致DNA损伤和凋亡。此外，胰腺炎的特征性病理反应，包括炎性趋化因子和僵硬的，脱落的微环境也可能有助于胰腺导管细胞中这种增加的氧化潜力。 ROS会引起严重的炎症反应，刺激了严重的疼痛，出血性休克，急性呼吸窘迫或死亡，但是，尚无体外人类模型可以理解这种病理。因此，我们开始开发一种新型，三维的人类胰管模型，该模型由Nestin阳性，永生的，胰腺上皮细胞，Matrigel支架，预定的培养基和特定生长因子组成。该模型是通过共焦显微镜来表征的，用于细胞组织，极化和功能，类似于人类乳腺上皮性acini。其次，在理解胰腺星状细胞对乙醇的反应方面取得了很多进展，但对胰腺上皮细胞反应的理解较少。 Thus, we will analyze the effect of increasing concentrations of ethanol on ROS induction within these human pancreatic ducts via fluorescent staining and lipid peroxidation as well as the subsequent DNA damage response and apoptotic initiation events via the double strand break markers (H2AX and (SP1101 and Caspase-3, Bim, and Annexin V, respectively. Thirdly, pancreatic stellate cells have also been然而，与炎症性介体的产生相关，在胰腺上皮细胞反应中几乎没有采用。最后。 By blocking pancreatic-specific integrin (3, (4 and (6 through antibodies/disintegrins and by DNA microarray validation of their expression, we will understand how exogenous elastic stiffness can affect ROS potential internal to pancreatic epithelial cells. We anticipate that the development of our in vitro, 3-D, human pancreatic epithelial duct model along with each of the three independent experiments will uncover parts of the病因学途径导致与人相关的模型中导致胰腺炎，并为将来的临床治疗提供了可能的靶标。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Electrospun hydroxyapatite-containing chitosan nanofibers crosslinked with genipin for bone tissue engineering.

DOI：
10.1016/j.biomaterials.2012.09.009
发表时间：
2012-12
期刊：
BIOMATERIALS
影响因子：
14
作者：
Frohbergh, Michael E.;Katsman, Anna;Botta, Gregory R.;Lazarovici, Phillip;Schauer, Caroline L.;Wegst, Ulrike G. K.;Lelkes, Peter I.
通讯作者：
Lelkes, Peter I.