Analysis of Radiomitigative Cell Signaling

放射抑制细胞信号传导分析

• 批准号: 8476331
• 负责人: GABOR J TIGYI
• 金额: $ 0.93万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-26 至 2013-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8476331
• 关键词: Adverse effects; Amino Acids; Apoptosis; Apoptotic; Applications Grants; Attenuated; BCL1 Oncogene; Back; C-terminal; Cells; Chimera organism; Coupled; DNA Damage; Development; Diarrhea; Disease; Dose; Edg4 Protein; Emergency Situation; Epithelial; Event; Explosion; Exposure to; Funding; G-Protein-Coupled Receptors; GTP-Binding Proteins; Growth Factor; Human; In Vitro; Knockout Mice; Leukocytes; Life; Ligands; Lysophosphatidic Acid Receptors; Lysophospholipids; MAPK3 gene; Mediating; Mediator of activation protein; Mitochondria; Molecular Target; Molecular Weight; Multiprotein Complexes; Mus; National Institute of Allergy and Infectious Disease; Nuclear Accidents; Patients; Phospholipids; Platelet Count measurement; Property; Protein Binding Domain; Proteins; Proto-Oncogene Proteins c-akt; Radiation; Radiation Injuries; Radiation Syndromes; Radiation therapy; Readiness; Research; Role; SRC gene; Signal Pathway; Signal Transduction; Site; Staging; Tail; Temperature; Tertiary Protein Structure; Testing; Therapeutic; United States National Institutes of Health; Work; analog; animal rule; attenuation; authority; base; chemotherapy; dirty bomb; irradiation; link protein; lysophosphatidic acid; mutant; new therapeutic target; novel; oxidative damage; protective effect; protein protein interaction; receptor; receptor coupling; research study; scaffold; sphingosine 1-phosphate; thiophosphate

DESCRIPTION (provided by applicant): Unintended exposure to radiation via a nuclear accident, or explosion of a "dirty bomb," can have devastating consequences. We developed a novel analog of the phospholipid growth factor lysophosphatidic acid (LPA), octadecenyl thiophosphate (OTP), which when applied up to 24h post-irradiation in mice irradiated with lethal doses of gamma-irradiation shows strong radiomitigating action by rescuing cells from apoptosis and saving life. Experiments indicate that the molecular target of OTP is the LPA2 G protein-coupled receptor. The objective of this application is to elucidate the radiomitigating mechanism of action unique to the LPA2 receptor. The central hypothesis of the proposed work is that macromolecular protein complexes formed at the C-terminal tail of the LPA2 receptor are required for the radioprotective effect of OTP. This is a significant departure from the concept that the LPA2 receptor coupled G protein-mediated signaling events are sufficient to elicit radiomitigation. LPA2 is distinctly different from the other LPA receptor subtypes in its C- terminal tail, where it forms protein-protein interactions with PDZ domain-binding proteins and with LIM domain proteins. We hypothesize that these LPA2-protein interactions regulate multiple prosurvival signaling pathways (ERK1/2 and AKT) and also arrest the actions Siva-1, a proapoptotic protein that links DNA damage to the apoptotic cascade. This hypothesis goes beyond the LPA2 GPCR because the sequence motifs responsible for these protein-protein interactions are present in many other GPCR. The specific aims are: 1. Test the hypothesis that the C-terminal PDZ motif is required for the radioprotective effect. 2. Characterize the motif in the C terminal 55 amino acids of the LPA2 receptor required for the interactions with the LIM domain proteins TRIP6 and Siva-1 by determining whether physical interaction with them is required for the antiapoptotic effect. 3. Dissect the role of LPA2-TRIP6 - c-Src - ERK/AKT signaling axis in the antiapoptotic effect. 4. Elucidate the role of the LPA2 - Siva-1- BCL-XL signaling axis in the antiapoptotic effect. An in-depth understanding of OTP's mechanism of action is of paramount importance for scientific, regulatory, and therapeutic reasons. Not negligible is the fact that OTP might also provide potential therapeutic applications in the attenuation of the side effects of radiation- and chemotherapy, as well as diseases associated with increased program cell death. Understanding the signaling interactions between pro-survival GPCR and the apoptotic machinery will pave the way to the identification of new therapeutic targets as well as GPCR ligands. Unintended exposure to radiation via a nuclear accident, or explosion of a "dirty bomb," can have devastating consequences to the public. We developed a novel analog of the phospholipid growth factor lysophosphatidic acid (LPA), octadecenyl thiophosphate (OTP). When applied up to 24 h post-irradiation in mice irradiated with lethal doses of gamma-irradiation, OTP shows significant radiomitigating action by rescuing cells from apoptosis and saving life. OTP has many of the properties of an ideal radioprotectant/radiomitigator: a) it is stable at room temperature for years; b) it can be formulated for a wide variety of patient types; c) it is relatively easy to synthesize and manufacture, and inexpensive; and d) it is effective when given up to 12 h after radiation injury. Experiments conducted in vitro and with knockout (KO) mice indicate that the molecular target of OTP is the LPA2 G protein-coupled receptor (GPCR). The objective of this grant application is to elucidate the an- tiapoptotic mechanism of action unique to the LPA2 receptor subtype

描述（由申请人提供）：意外暴露于核事故或“肮脏炸弹”的爆炸可能会带来毁灭性后果。我们开发了一种新颖的类似物，对磷脂生长因子溶物磷酸酸（LPA），甲氧基甲基磷酸盐（OTP）进行了新的类似物，该磷脂酸（OTP）在小鼠后24小时施用时，施加了24小时，该小鼠的辐照剂施加了γ-iRradiation的致命剂量，通过弹药可通过挽救Apoptosis的弹药率挽救细胞，并挽救了强烈的放射性动作。实验表明OTP的分子靶标是LPA2 G蛋白偶联受体。该应用的目的是阐明LPA2受体独特的作用机理。所提出的工作的中心假设是在LPA2受体的C末端形成的大分子蛋白复合物是OTP的辐射保护作用所必需的。这与LPA2受体耦合G蛋白介导的信号传导事件的概念具有很大的不同，足以引起放射线。 LPA2与其C末端尾部中的其他LPA受体亚型明显不同，在该尾部，它与PDZ结构域结合蛋白和LIM结构域蛋白质形成蛋白质 - 蛋白质相互作用。我们假设这些LPA2-蛋白质相互作用调节了多个Proservival信号通路（ERK1/2和AKT），并且也阻止了将DNA损伤与凋亡级联的促凋亡蛋白SIVA-1的作用。该假设超出了LPA2 GPCR，因为在许多其他GPCR中都存在负责这些蛋白质蛋白相互作用的序列基序。具体目的是：1。检验以下假设：辐射保护作用需要C末端PDZ基序。 2。通过确定是否需要与抗凋亡作用的物理相互作用来表征与LIM结构域蛋白Trip6和Siva-1相互作用所需的LPA2受体55氨基酸的基序。 3。剖析LPA2 -TRIP6 -C -SRC -ERK/AKT信号轴在抗凋亡效应中的作用。 4。阐明LPA2-SIVA-1-BCL-XL信号轴在抗凋亡效应中的作用。对OTP的作用机理的深入了解对于科学，调节和治疗原因而言至关重要。 OTP可能还可以在放射线和化学疗法的副作用以及与程序细胞死亡增加有关的疾病中提供潜在的治疗应用。了解促生存GPCR和凋亡机制之间的信号传导相互作用将为鉴定新的治疗靶标和GPCR配体铺平道路。意外通过核事故接触辐射，或者“肮脏的炸弹”的爆炸可能对公众产生毁灭性的后果。我们开发了磷脂生长因子溶物磷脂酸（LPA），甲苯甲基磷酸盐（OTP）的新型类似物。当在用致死剂量的γ-辐照辐照的小鼠中施用多达24小时的射精后，OTP通过挽救细胞中的细胞凋亡和挽救生命而显示出明显的放射性作用。 OTP具有理想放射治疗剂/放射线仪的许多特性：a）在室温下多年的稳定； b）可以针对多种患者类型制定； c）相对容易合成和制造且价格便宜； d）当放射线损伤后12小时允许时，它是有效的。在体外和敲除（KO）小鼠中进行的实验表明，OTP的分子靶标是LPA2 G蛋白偶联受体（GPCR）。该赠款应用的目的是阐明LPA2受体亚型独有的作用机理

项目成果

Design and synthesis of sulfamoyl benzoic acid analogues with subnanomolar agonist activity specific to the LPA2 receptor.

• DOI: 10.1021/jm5007116
• 发表时间: 2014-08-28
• 期刊: Journal of medicinal chemistry
• 影响因子: 7.3
• 作者: Patil R;Fells JI;Szabó E;Lim KG;Norman DD;Balogh A;Patil S;Strobos J;Miller DD;Tigyi GJ
• 通讯作者: Tigyi GJ

In honor of Robert Bittman 1942-2014.

纪念 1942 年至 2014 年的罗伯特·比特曼。

• DOI: 10.1016/j.bbalip.2014.10.005
• 发表时间: 2014
• 期刊: Biochimica et biophysica acta
• 作者: Tigyi,Gabor