Theta-defensins Novel HIV-1 Uptake Inhibitors

Theta-防御素新型 HIV-1 摄取抑制剂

• 批准号: 7217903
• 负责人: ROBERT IRVING LEHRER
• 金额: $ 25.64万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-05-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7217903
• 关键词: Affinity; Amino Acids; Animals; Anti-Retroviral Agents; Antiviral Agents; Appendix; Arts; Basic Science; Binding; Biological Assay; Bone Marrow; CD4 Positive T Lymphocytes; Carbohydrates; Cell Line; Cells; Cercopithecidae; Cervical; Chemistry; Cyanovirin-N; Defensins; Dendritic Cells; Development; Endopeptidases; Epithelial; Epithelial Cells; Future; Genes; Grant; HIV Envelope Protein gp120; HIV-1; Health; Heating; Hominidae; Host Defense; Host resistance; Human; Human Genome; Human Herpesvirus 2; Immune; Immune system; In Vitro; Infection; Interferons; Lactobacillus; Lead; Lectin; Leukocytes; Macaca mulatta; Messenger RNA; Molecular; Mucous Membrane; Mutation; Nonsense Codon; Oligosaccharides; Pathogenesis; Pattern; Peptide Hydrolases; Peptide Synthesis; Peptides; Peripheral Blood Mononuclear Cell; Pharmacologic Substance; Phase; Polysaccharides; Primates; Property; Research; Research Design; Research Personnel; Resistance; Retroviridae; Role; Solid; Specificity; Spleen; Structure; Surface Plasmon Resonance; T-Lymphocyte; Techniques; Testing; Therapeutic; Thymus Gland; Topical agent; Translations; Vagina; Viral; alpha-Defensins; analog; base; carbohydrate structure; cytotoxicity; in vivo; inhibitor/antagonist; killings; member; microbial; monocyte; nonhuman primate; novel; pathogen; prevent; retrocyclin; sugar; synthetic peptide; tandem mass spectrometry; theta-defensin; uptake; vaginal lactobacilli

DESCRIPTION (provided by applicant): We propose to study the immune mechanism of defensins in host resistance to human HIV-1 and related retroviruses of non-human primates. Our long-term objectives are: 1) to identify innate immune system molecules involved in the pathogenesis of HTV-1 infections in primates. 2) to characterize the carbohydrate-binding specificity of theta- and alpha-defensins, and to correlate this property with their activity against HIV-1. 3) to identify potent theta-defensins for future in vivo studies designed to assess their therapeutic usefulness. No in vivo studies are proposed hi this grant. Theta-defensins are cyclic octadecapeptides expressed by leukocytes of Old World monkeys and some other non human primates. Their ability to recognize certain carbohydrate structures (glycans) allows some 9-defensins to bind both gp120 of HIV-1 and CD4 with high affinity, and to prevent the entry of R5 and X4 strains of HIV-1 into otherwise susceptible human cells. At least 6 0-defensin genes exist in the human genome, and theta-defensin mRNA exists in human bone marrow, thymus, and spleen. However, human theta-defensin genes and their mRNA contain a premature stop codon, and human cells do not produce theta-defensin peptides. Retrocyclins 1-3 are synthetic peptides that represent the theta-defensins that humans could produce if the silencing mutation (the premature stop codon) were absent. Retrocyclin-2 is effective against HIV-1 primary isolates representing subtypes A,B,C and CRF01_AE, which cause most HIV-1 infections worldwide. Because retrocyclins are noncytotoxic and noninjurious to vaginal lactobacilli, they provide intriguing lead compounds for future pharmaceutical development. The studies contained in this proposal are intended to provide a knowledge-platform that will facilitate the development of therapeutic theta-defensins in the future. The specific aims are: 1). To synthesize novel theta-defensins, including peptides based on theta-defensin gene sequences in non-human primates and analogues of human retrocyclins, and to test them for activity against HIV-1 and cytotoxicity. 2). To identify the sugars and oligosaccharides that primate theta-defensins bind, and correlate binding with their antiretroviral properties. Aim 1 will be accomplished by solid phase peptide synthesis and in vitro protection assays using peripheral blood mononuclear cells and indicator cell lines. Specific Aim 2 will be realized by a combination of microchemical techniques, including surface plasmon resonance, tandem mass spectrometry, and state-of-the-art lectinomic and peptidomic chemistry.

描述（由申请人提供）：我们建议研究防御素在宿主对人HIV-1的抗性和非人类灵长类动物的相关逆转录病毒中的免疫机制。我们的长期目标是：1）确定参与灵长类动物中HTV-1感染发病机理的先天免疫系统分子。 2）表征硫代和α-防御素的碳水化合物结合特异性，并将其与HIV-1的活性相关联。 3）鉴定有效的硫代抗素，以供未来的体内研究，旨在评估其治疗用途。没有提出本体内研究。 theta-defensin是由旧世界猴子和其他一些非人类灵长类动物的白细胞表达的环状八肽。它们识别某些碳水化合物结构（Glycans）的能力允许一些9-防御素结合具有高亲和力的HIV-1和CD4的GP120，并防止HIV-1的R5和X4菌株进入其他易感人类细胞。人类基因组中至少存在至少6个0-防御素基因，硫素 - 德防御素mRNA存在于人骨髓，胸腺和脾脏中。然而，人抑制抑制素基因及其mRNA含有过早的终止密码子，并且人类细胞不产生抑制抑制肽的肽。腹膜循环蛋白1-3是代表抑制抑制素的合成肽，如果没有沉默突变（过早的终止密码子），则可以产生人类。逆转录蛋白2对代表亚型A，B，C和CRF01_AE的HIV-1主要分离株有效，这会引起全球大多数HIV-1感染。由于腹胞菌蛋白是非胞毒性的，并且对阴道乳酸杆菌不舒适，因此它们为未来的药物开发提供了有趣的铅化合物。本提案中包含的研究旨在提供一个知识平台，将来将促进治疗性theta-defensin的发展。具体目的是：1）。为了合成新型抑制抑制素，包括基于非人类灵长类动物和人逆转录蛋白的类似物中抑制抑制素基因序列的肽，并测试它们是否针对HIV-1和细胞毒性。 2）。确定灵长类动物抑制素结合的糖和寡糖，并与其抗逆转录病毒特性相关。 AIM 1将通过使用外周血单核细胞和指示细胞系的固相肽合成和体外保护测定来实现。特定目标2将通过微化学技术（包括表面等离子体共振，串联质谱和最新的凝集型和肽组化学）的组合来实现。