CEHS Summer Undergraduate Research Program: University of Montana

CEHS 夏季本科生研究计划：蒙大拿大学

• 批准号: 10195323
• 负责人: Andrij Holian
• 金额: $ 12.5万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10195323
• 关键词: Acrolein; Administrative Supplement; Adverse effects; Agonist; Alveolar Macrophages; Arginine; Biological; Biology; Cations; Chemicals; Chemistry; Critical Pathways; Cysteine; DNA; Data; Disease; Dose; Epithelial Cells; Eye; Eye Injuries; Family; Funding; Future; Glutathione; Goals; Guanine; Histidine; Human; In Vitro; Inflammatory; Inhalation; Intervention; Knowledge; Laboratories; Literature; Lung; Lysine; Molecular Target; Montana; NF-kappa B; Outcome; PTPN1 gene; Pathway interactions; Peptide Hydrolases; Permeability; Phenotype; Phosphoric Monoester Hydrolases; Production; Protein Tyrosine Phosphatase; Proteins; Reporting; Research; Role; Site; Skin; Skin injury; Specificity; Sulfhydryl Compounds; Testing; Therapeutic Intervention; Time; Toxic effect; Toxicology; Translating; Universities; adverse outcome; base; chemical threat; cytokine; experience; genetic regulatory protein; in vivo; innate immune function; macrophage; medical countermeasure; member; prevent; programs; receptor; research and development; respiratory; response; sex; src-Family Kinases; transcription factor; undergraduate research

Acrolein (CH2=CHCHO) is a highly-reactive, volatile chemical that may be released accidentally or intentionally, resulting in eye, skin, and respiratory toxicity. Based on acrolein’s chemistry, it is known to target biological nucleophiles including guanine in DNA, as well as cysteine, lysine, histidine and arginine residues in critical regions of transcription factors, proteases and other proteins. Consequently, with such a broad range of potential biological targets it has been difficult to be certain of the most sensitive targets in vivo on which to base possible therapeutic interventions. Acrolein has been reported to be an agonist for the Ca2+-permeable transient receptor potential cation channel, subfamily A, member 1 (TRPA1). Therefore, acrolein targeting TRPA1 on lung macrophages would be expected to increase inflammatory cytokine production. However, studies have reported that acrolein inhibits macrophage cytokine production. Acrolein is expected to target thiol groups that are common critical sites for activity of protein tyrosine phosphatases (PTP) making them an important and logical family of target proteins that would explain inhibition of macrophage cytokine production. Of the various PTP, PTP1B has been reported to be irreversibly inhibited by acrolein, but there are no reports that PTP1B is a target of acrolein toxicity in vivo. Since PTP1B is a key regulatory PTP involved in critical pathways and diseases and as a target of acrolein exposure could explain a number of the reported outcomes of acrolein exposure especially at sublethal concentrations. Since the precise mechanism of acrolein toxicity remains uncertain, this project will fill an important gap of knowledge to define in vitro and demonstrate for the first time in vivo, whether inhibition of PTP1B catalytic activity helps define the mechanism of acrolein-induced respiratory toxicity at sublethal concentrations. Studies in Aim 1 will Demonstrate that acrolein blocks the activity of PTP1B in macrophages and epithelial cells in vitro and will be compared to acrolein’s effects on TRPA1. Studies in Aim 2 will demonstrate that sublethal in vivo exposures of acrolein blocks the activity of PTP1B in lung macrophages to decrease inflammatory cytokine release from macrophages. These studies will provide new mechanistic information on acrolein toxicity in vitro and in vivo that will help explain the reported adverse impacts of acrolein exposure and provide guidance to develop medical countermeasures to alleviate the adverse effects of acrolein exposures.

丙烯醛（CH 2 =CHCHO）是一种高活性、挥发性化学品，可能会意外或故意释放， 导致眼睛、皮肤和呼吸道中毒。基于丙烯醛的化学性质，已知其靶向生物 亲核试剂包括DNA中的鸟嘌呤，以及关键氨基酸中的半胱氨酸、赖氨酸、组氨酸和精氨酸残基。 转录因子、蛋白酶和其他蛋白质的区域。因此，有了如此广泛的潜力， 生物学靶点很难确定体内最敏感的靶点， 治疗干预。据报道丙烯醛是Ca2+渗透性瞬时受体的激动剂 潜在阳离子通道，亚家族A，成员1（TRPA1）。因此，丙烯醛靶向肺上的TRPA 1， 预期巨噬细胞会增加炎性细胞因子的产生。然而，研究报告称， 丙烯醛抑制巨噬细胞细胞因子的产生。预期丙烯醛靶向硫醇基团， 蛋白酪氨酸磷酸酶（PTP）活性的共同关键位点使其成为重要的和合乎逻辑的 这是一个靶蛋白家族，可以解释巨噬细胞细胞因子产生的抑制。在各种PTP中， PTP1B被丙烯醛不可逆地抑制，但没有报道PTP1B是丙烯醛的靶点 丙烯醛的毒性。由于PTP1B是参与关键途径和疾病的关键调节性PTP， 作为丙烯醛暴露的目标可以解释丙烯醛暴露的许多报告结果， 在亚致死浓度。由于丙烯醛毒性的确切机制仍然不确定，该项目将 填补了一个重要的知识空白，在体外定义并首次在体内证明， PTP1B催化活性有助于确定丙烯醛诱导的亚致死呼吸毒性机制 浓度的目的1中的研究将证明丙烯醛阻断巨噬细胞中PTP1B的活性， 上皮细胞体外培养，并将其与丙烯醛对TRPA1的影响进行比较。目标2中的研究将证明 丙烯醛的亚致死体内暴露阻断肺巨噬细胞中PTP1B的活性， 从巨噬细胞释放炎性细胞因子。这些研究将提供新的机制信息， 体外和体内丙烯醛毒性，这将有助于解释丙烯醛暴露的不良影响， 为制定医疗对策提供指导，以减轻丙烯醛暴露的不利影响。