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（申请人提供）：我们建议研究防御素在宿主抵抗人类HIV-1和非人类灵长类相关逆转录病毒中的免疫机制。我们的长期目标是：1) 鉴定与灵长类动物 HTV-1 感染发病机制有关的先天免疫系统分子。 2) 表征 θ 和 α 防御素的碳水化合物结合特异性，并将这一特性与其抗 HIV-1 的活性相关联。 3）确定有效的θ防御素，用于未来旨在评估其治疗有效性的体内研究。本次资助未提出体内研究。 θ-防御素是由旧大陆猴子和其他一些非人类灵长类动物的白细胞表达的环状十八肽。它们识别某些碳水化合物结构（聚糖）的能力使得一些 9-防御素能够以高亲和力结合 HIV-1 的 gp120 和 CD4，并阻止 HIV-1 的 R5 和 X4 毒株进入其他易感的人类细胞。人类基因组中至少存在6个0-防御素基因，θ-防御素mRNA存在于人类骨髓、胸腺和脾脏中。然而，人类θ防御素基因及其mRNA含有提前终止密码子，并且人类细胞不产生θ防御素肽。逆转录环素 1-3 是合成肽，代表如果沉默突变（过早终止密码子）不存在，人类可以产生的 θ 防御素。 Retrocyclin-2 可有效对抗代表 A、B、C 和 CRF01_AE 亚型的 HIV-1 原发分离株，这些亚型是全球大多数 HIV-1 感染的原因。由于逆转录素对阴道乳酸杆菌无细胞毒性和无害性，因此它们为未来的药物开发提供了有趣的先导化合物。该提案中包含的研究旨在提供一个知识平台，以促进未来治疗性θ防御素的开发。具体目标是：1）．合成新型 θ 防御素，包括基于非人类灵长类动物 θ 防御素基因序列的肽和人类逆转录素类似物，并测试它们的抗 HIV-1 活性和细胞毒性。 2）。鉴定灵长类动物θ防御素结合的糖和寡糖，并将结合与其抗逆转录病毒特性相关联。目标 1 将通过固相肽合成和使用外周血单核细胞和指示细胞系的体外保护测定来实现。具体目标 2 将通过结合微化学技术来实现，包括表面等离子体共振、串联质谱以及最先进的电子组学和肽组学化学。