The Role of Scavenger Receptor gp340 in Mucosal HIV-1 Transmission and Inhibition

清道夫受体 gp340 在粘膜 HIV-1 传播和抑制中的作用

• 批准号: 8743609
• 负责人: Min Lu
• 金额: $ 49.23万
• 依托单位: RBHS-NEW JERSEY MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-23 至 2016-01-22
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8743609
• 关键词: Acquired Immunodeficiency Syndrome; Address; Administrative Supplement; Affinity; Antiviral Agents; Binding; Binding Sites; Biological Assay; CCR5 gene; CD209 gene; CD4 Positive T Lymphocytes; Cell Line; Cell fusion; Cell surface; Cell-Matrix Junction; Cells; Cervical; Chinese Hamster Ovary Cell; Collaborations; Competence; Complement; Complex; Dentistry; Development; Disease; Effectiveness; Epithelial; Epithelium; Failure; Female; Funding; Genital system; Glycoproteins; Goals; Grant; HIV Envelope Protein gp120; HIV-1; Human; Hurricane; Immunoglobulin Variable Region; In Vitro; Infection; Inhibitory Concentration 50; Intervention; Investigation; Knowledge; Lead; Membrane Fusion; Modeling; Molecular; Molecular Target; Molecular Virology; Multi-Drug Resistance; Mus; Mutation; New Jersey; New York; North Carolina; Oral; Oryctolagus cuniculus; Peptides; Pharmaceutical Preparations; Prevalence; Process; Production; Property; Proteins; Reagent; Receptor Cell; Recombinants; Research; Research Project Grants; Role; SRCR proteins; Saliva; Specificity; Structure; Surface; T-Lymphocyte; Testing; Tissues; Toxic effect; Universities; Vagina; Variant; Viral; Virus; base; biophysical chemistry; biophysical techniques; college; combat; cost; design; disulfide bond; env Glycoproteins; in vitro Model; in vivo; inhibitor/antagonist; insight; macrophage; medical schools; neutralizing antibody; neutralizing monoclonal antibodies; novel; parent grant; polyclonal antibody; prevent; programs; receptor binding; reproductive; research study; scavenger receptor; structural biology; syndecan; synthetic peptide; transcytosis; transmission process; vaginal transmission; virucide

The research outlined in this Administrative Supplement to Grant R01 AI094599 involves an ongoing collaboration between Min Lu (Rutgers New Jersey Medical School) and Daniel Malamud (New York University College of Dentistry). Due to power failure during Hurricane Sandy, we lost all stable Chinese hamster ovary (CHO) cell lines expressing SRCR1 module fragments of the human scavenger receptor gp340, polyclonal antibodies raised in rabbits against a synthetic peptide derived from the SRCR1 module, and gp340 peptide and protein entry inhibitors. The supplemental funds are requested to reproduce these critical and proprietary research materials and reagents and to define the molecular target and mechanism of action of soluble gp340- based anti-HIV-1 agents. These studies could lead to discovery of novel entry inhibitors that target early infection to prevent mucosal HIV-1 transmission. Experiment 1: To generate stable CHO cell lines for high-level production of secreted SRCR1 module fragments. Experiment 2: To characterize the structural properties of the SRCR1 domain both free and bound to the V3 region of HIV-1 gp120. Our overall goal is to elucidate how HIV-1 envelope glycoprotein function in cell entry is modulated through the gp340-gp120 interaction; to use this knowledge to optimize SRCR1 entry inhibitors for antiviral potency; and to test these hypotheses in a humanized mouse vaginal transmission model, in collaboration with Dr. J. Victor Garcia-Martinez (University of North Carolina School of Medicine) in years 4 and 5. To achieve this goal, we will express and purify large quantities of recombinant gp340 fragments in CHO cells, and carry out a combined structural and physicochemical investigation of the SRCR1 domain in complex with the V3 region. These experiments will enable us to develop optimized SRCR1 entry inhibitors for evaluating their effectiveness to protect against mucosal HIV-1 infection in vivo. Fulfilling the central objective of the parent grant would not be possible without the with-cost extension of support that would significantly mitigate the impact of Sandy on this research project.

本行政补充拨款R 01 AI 094599中概述的研究涉及正在进行的 闵陆（Rutgers新泽西医学院）和丹尼尔马拉默德（纽约大学）之间的合作 牙科学院）。由于桑迪飓风期间停电，我们失去了所有稳定的中国仓鼠卵巢 (CHO)表达人清道夫受体gp 340的SRCR 1模块片段的多克隆细胞系 在兔中产生的针对源自SRCR 1模块的合成肽和gp 340肽的抗体 和蛋白质进入抑制剂。补充资金用于复制这些关键和专有的 研究材料和试剂，并确定可溶性gp 340- 抗HIV-1药物。这些研究可能会导致发现新的进入抑制剂，靶向早期 感染，以防止粘膜HIV-1传播。 实验1：生成用于高水平生产分泌型SRCR 1模块的稳定CHO细胞系 片段 实验2：为了表征SRCR 1结构域的结构特性，包括游离的和结合的SRCR 1结构域， HIV-1 gp 120的V3区。 我们的总体目标是阐明HIV-1包膜糖蛋白在细胞进入中的功能是如何通过 gp 340-gp 120相互作用;使用该知识优化SRCR 1进入抑制剂的抗病毒效力;以及 与J.维克托博士合作，在人源化小鼠阴道传播模型中检验这些假设 Garcia-Martinez（北卡罗来纳州医学院大学）在4年和5年。为了实现这一目标，我们 将在CHO细胞中表达和纯化大量重组gp 340片段，并进行 结合SRCR 1结构域与V3区复合的结构和物理化学研究。 这些实验将使我们能够开发优化的SRCR 1进入抑制剂，用于评估其生物学特性。 有效地防止体内粘膜HIV-1感染。实现母公司的中心目标 如果不扩大支持范围（这将大大减轻 桑迪对本研究项目的影响。