Regulation of the baboon HLA-G-like class Ib MHC gene, Paan-AG

狒狒 HLA-G 样 Ib 类 MHC 基因 Paan-AG 的调控

• 批准号: 7300396
• 负责人: DAUDI K LANGAT
• 金额: $ 7.35万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7300396
• 关键词: 3' Untranslated Regions; Alleles; Allogenic; Alternative Splicing; Animal Model; Binding; Biological Assay; Cell Line; Cells; Characteristics; Class; Code; Cytoplasmic Tail; Data; Development; Disease; Drug or chemical Tissue Distribution; Electrophoretic Mobility Shift Assay; Elements; Embryo; Embryo Transfer; Embryonic Development; Ensure; Fertilization; Fertilization in Vitro; Fetus; Future; GTP-Binding Proteins; Gametogenesis; Genes; Genetic Polymorphism; Genetic Transcription; Goals; HLA Antigens; HLA-A Antigens; HLA-A gene; HLA-A2 Antigen; HLA-G5; HLA-G7 antigen; Health; Heart Transplantation; Homologous Gene; Human; Immune; Immune system; Immunosuppressive Agents; In Vitro; Indium; Initiator Codon; Interferons; Introns; Laboratories; Location; Luciferases; Manuscripts; Maternal-Fetal Exchange; Mediating; Membrane; Messenger RNA; Modeling; Numbers; Oocytes; Papio; Papio anubis; Pathology; Phenotype; Placenta; Placentation; Pre-Eclampsia; Pregnancy; Pregnancy Complications; Pregnancy loss; Primates; Process; Production; Protein Isoforms; Proteins; RNA; RNA Splicing; Recombinants; Regulation; Regulatory Element; Regulatory Pathway; Reporter; Reproductive system; Research; Research Personnel; Role; Site; Syncytiotrophoblast; Testing; Therapeutic; Therapeutic Uses; Thinking; Tissues; Transcript; Transgenic Mice; Translations; Transplantation; Untranslated Regions; base; blastocyst; cytotrophoblast; design; hydroxy-aluminum polymer; implantation; in vitro Model; in vivo; in vivo Model; insight; mRNA Expression; programs; promoter; protein expression; research study; software systems; success; transcription factor; tumor; zygote

DESCRIPTION (provided by applicant): The human leukocyte antigen-G (HLA-G), a protein highly expressed at the human maternal- fetal interface during pregnancy, is thought to be critical in survival of the semi-allogenic fetus. Our long-term goal is to elucidate the role of HLA-G in pregnancy as a prerequisite to assessing the therapeutic potential of recombinant HLA-G proteins in pregnancy-related pathologies and transplantation. Current evidence suggests HLA-G programs immune cells at the maternal-fetal interface into immunosuppressive phenotypes. It has been implicated in development of preeclampsia, implantation success in in vitro fertilization (IVF) programs, survival of heart transplants and survival of some tumors. However, definitive proof of HLA-G function remains elusive since in vivo experiments in humans are not possible due to ethical concerns. In the search for an appropriate animal model, we have identified the olive baboon (Papio anubis) as a potential candidate. This primate expresses an HLA-G-like protein termed Paan-AG in the placenta. Our hypothesis is that Paan-AG could be the functional homologue of HLA-G. Preliminary data shows that Paan-AG shares many characteristics with HLA-G, including limited polymorphism, alternative splicing of the mRNA, and restricted tissue of expression of the protein (mainly in the placenta). The restricted tissue expression of the proteins suggests the two genes might share tissue-specific regulatory elements. In order to confirm whether the baboon is a useful model, it is necessary to establish whether regulation of Paan-AG is similar to that of HLA-G as known to date. The aim of this proposal is to identify tissue-specific regulatory elements in the 5' and 3' untranslated regions or introns of Paan-AG. We previously identified a number of putative regulatory elements in these regions using a transcription factor search system software. We will assess activity of these elements using electrophoretic mobility shift assays and luciferase reporter assays in different cell lines. The results will show whether the tissue-specific expression of Paan-AG is controlled by tissue- specific regulatory element, and the location of such element(s). A negative result will guide our future research to focus on post-translational mechanisms as candidates for tissue-specific expression of Paan-AG, and by inference, HLA-G. TO PUBLIC HEATH: Understanding the function of HLA-G is critical because of the potential therapeutic use of recombinant HLA-G proteins in the management of pregnancy complications or in transplantation. The baboon's reproductive system closely resembles that of human in terms of maturation sequence of fertilized eggs, implantation and placentation process, and in production of HLA-G- like proteins (Paan-AG) in the placenta. Thus the baboon is clearly an excellent model for HLA-G functional experiments, and testing the therapeutic applications of HLA-G in human pregnancy and transplantation.

描述（由申请人提供）：人白细胞抗原- g （HLA-G）是妊娠期间在人母胎界面高度表达的一种蛋白，被认为对半同种异体胎儿的存活至关重要。我们的长期目标是阐明HLA-G在妊娠中的作用，作为评估重组HLA-G蛋白在妊娠相关病理和移植中的治疗潜力的先决条件。目前的证据表明，HLA-G程序免疫细胞在母胎界面进入免疫抑制表型。它与先兆子痫的发展、体外受精（IVF）计划的植入成功、心脏移植的存活和一些肿瘤的存活有关。然而，HLA-G功能的明确证据仍然难以捉摸，因为由于伦理问题，不可能在人类体内进行实验。在寻找合适的动物模型时，我们已经确定了橄榄狒狒（Papio anubis）作为潜在的候选者。这种灵长类动物在胎盘中表达一种称为Paan-AG的hla - g样蛋白。我们的假设是Paan-AG可能是HLA-G的功能同源物。初步数据显示Paan-AG与HLA-G具有许多共同的特征，包括有限的多态性，mRNA的选择性剪接以及蛋白质表达的限制性组织（主要在胎盘中）。这两种蛋白在组织中的限制性表达表明，这两个基因可能共享组织特异性调控元件。为了确认狒狒是否是一个有用的模型，有必要确定Paan-AG的调节是否与目前已知的HLA-G相似。本提案的目的是鉴定Paan-AG的5‘和3’非翻译区或内含子中的组织特异性调控元件。我们之前使用转录因子搜索系统软件确定了这些区域的一些假定的调控元件。我们将在不同的细胞系中使用电泳迁移率转移测定和荧光素酶报告基因测定来评估这些元素的活性。结果将显示Paan-AG的组织特异性表达是否受到组织特异性调控元件的控制，以及这些元件的位置。阴性结果将指导我们未来的研究重点放在作为Paan-AG组织特异性表达候选物的翻译后机制上，由此推断，HLA-G。公共卫生：了解HLA-G的功能是至关重要的，因为重组HLA-G蛋白在妊娠并发症管理或移植中的潜在治疗用途。狒狒的生殖系统在受精卵的成熟顺序、着床和胎盘过程以及胎盘中HLA-G样蛋白（Paan-AG）的产生方面与人类非常相似。因此，狒狒显然是HLA-G功能实验的优秀模型，并测试HLA-G在人类妊娠和移植中的治疗应用。