Single Molecule Studies of HIV Envelope Properties

HIV 包膜特性的单分子研究

• 批准号: 8232033
• 负责人: Krishanu Ray
• 金额: $ 11.12万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8232033
• 关键词: Anti-Retroviral Agents; Antibodies; Award; Binding; Biological; Biological Assay; Biology; Biomedical Research; CD4 Antigens; Cell Surface Receptors; Cell surface; Cells; Characteristics; Complement; Complex; Core Protein; Data; Detection; Development; Dissociation; Educational Curriculum; Enzymes; Epitopes; Event; Fluorescence; Fluorescence Anisotropy; Fluorescence Resonance Energy Transfer; Fluorescence Spectroscopy; Goals; HIV; HIV Envelope Protein gp120; HIV Infections; HIV-1; Human; Human Virology; Immobilization; In Vitro; Individual; Infection; Institutes; K-Series Research Career Programs; Knowledge; Learning; Link; Location; Longevity; Macromolecular Complexes; Maryland; Masks; Measurement; Measures; Mediating; Membrane; Mentors; Mentorship; Methods; Molecular; Molecular Cloning; Molecular Conformation; Monoclonal Antibodies; Nature; Pharmaceutical Preparations; Physics; Population Heterogeneity; Predisposition; Process; Property; Proteins; Research; Research Personnel; Research Training; Resources; Sampling; Scientist; Solid; Spectrum Analysis; Speed; Staging; Structural Models; Structure; Surface; Techniques; Thermodynamics; Time; Training; Universities; Vaccines; Viral; Virion; Virus; Weight; antigen binding; base; career; conformer; drug development; env Glycoproteins; experience; flexibility; human monoclonal antibodies; inhibitor/antagonist; medical schools; multidisciplinary; neutralizing antibody; novel; optical imaging; prevent; programs; receptor; receptor binding; research study; single molecule; skills; spectroscopic imaging; stoichiometry; three dimensional structure; vaccine development; virus host interaction

DESCRIPTION (provided by applicant): The overall objective of the proposed Mentored Quantitative Research Career Development Award (K25) is to provide a period of mentored didactic and research training that will allow Dr. Krishanu Ray with his physics background to make a career transition into an independent biomedical researcher. The immediate goal is for Dr. Ray to expand his current research program in applying advanced fluorescence spectroscopy techniques to interrogate virus-host interactions in early stages of HIV infection. Specifically, Dr. Ray will employ novel fluorescence based approaches not previously described for viral entry process. The four-year training plan involves a structured curriculum of coursework and seminars focusing on HIV related research. This program will be heavily weighted toward gaining expertise regarding the HIV biology, particularly using optical imaging and spectroscopy based approaches to HIV research under the mentorship of two exceptional scientists, Dr. Joseph R. Lakowicz and Dr. Anthony L. DeVico at the University of Maryland School of Medicine (UMSM). Because of Dr. Ray's location in the Center for Fluorescence Spectroscopy (CFS) and adjacent to the Institute of Human Virology (IHV) at the UMSM, it is possible to study the biophysical properties of HIV proteins and neutralizing antibody binding against HIV-1 envelope. The long-term goal is for Dr. Ray to acquire advanced biomedical research skills and knowledge that complement his skills as a spectroscopist. The ultimate goal is for him to develop into an independent, interdisciplinary biomedical researcher. This K25 award will enable Dr. Ray to take advantage of the outstanding resources available at CFS and IHV and expand his expertise and experience in this unique multidisciplinary setting. Dr. Ray will develop methods applying fluorescence correlation spectroscopy (FCS) and single molecule detection (SMD) to directly probe the nature of HIV-1 envelope interactions with cell surface receptors and/or anti-envelope antibodies at the molecular level. It is hypothesized that a detailed understanding of the conformational dynamics that impact the HIV envelope before and during receptor engagement will explain the observed susceptibility to various inhibitors. Accordingly, Dr. Ray will focus on the following aims: 1) To develop methods for examining HIV-1 gp120-CD4 interactions at the single molecule level. 2) To characterize HIV-1 Env trimer conformational dynamics using single molecule spectroscopy. 3) To compare the binding of human monoclonal antibodies with different neutralizing potencies to HIV-1 envelope trimers by FCS and SMD. Successful completion of the Aims will provide new quantitative approaches at the individual molecular level towards understanding early steps in viral entry. This program will provide an excellent milieu in which the applicant - under close mentorship - can become deeply involved in applying advanced fluorescence spectroscopic and imaging approaches to HIV research. Mentorship will be geared more toward biological issues to provide a well-rounded portfolio as the candidate progresses toward independence in biomedical spectroscopic and imaging research. Human-immunodeficiency virus type 1 (HIV-1) has been the subject of intensive research. The majority of antiretroviral drugs acts at a post-infection step and cannot cure HIV-1 infection. Consequently, there is a continued emphasis on developing either drugs or vaccines that block pre-entry events by targeting the HIV-1 surface trimers which enable binding to cell surface coreceptors. In this application, the nature of HIV-1 envelope interactions with cell surface receptors and/or anti-envelope antibodies will be determined at the single molecule level. This novel application of single molecule fluorescence based approaches is expected to provide unique information relevant to vaccine and drug development.

描述（由申请人提供）：拟议的指导定量研究职业发展奖（K25）的总体目标是提供一段指导的教学和研究培训，这将使Krishanu Ray博士具有其物理背景，以使职业过渡到独立的生物医学研究人员。直接的目标是Ray博士扩大他当前的研究计划，以应用高级荧光光谱技术来询问HIV感染的早期阶段病毒宿主相互作用。具体而言，Ray博士将采用先前未针对病毒进入过程描述的新型基于荧光的方法。为期四年的培训计划涉及课程的结构化课程和针对HIV相关研究的研讨会。该计划将在获得HIV生物学方面的专业知识方面受到重大加权，尤其是在两位杰出科学家Joseph R. Lakowicz博士和马里兰大学医学院（UMSM）的两位杰出科学家Joseph R. Lakowicz和Anthony L. Devico博士的指导下，使用基于光学成像和基于光谱研究的HIV研究方法。由于Ray博士在UMSM的人类病毒学研究所（IHV）附近的荧光光谱中心（CFS）的位置，因此可以研究HIV蛋白的生物物理特性并中和抗体与HIV-1信封的结合。长期的目标是让雷博士获得高级生物医学研究技能和知识，以补充他作为光谱学家的技能。最终的目标是使他发展成为独立的跨学科生物医学研究人员。这项K25奖将使Ray博士能够利用CFS和IHV的出色资源，并在这种独特的多学科环境中扩大他的专业知识和经验。 Ray博士将开发用于应用荧光相关光谱（FCS）和单分子检测（SMD）的方法，以直接探测HIV-1 Invelope与细胞表面受体和/或抗Envelope抗体的性质。假设对影响受体参与之前和期间影响HIV包膜的构象动力学的详细理解将解释观察到的对各种抑制剂的敏感性。因此，Ray博士将重点关注以下目的：1）开发用于检查单分子水平上HIV-1 GP120-CD4相互作用的方法。 2）使用单分子光谱法表征HIV-1 Env Trimer构象动力学。 3）比较了人类单克隆抗体与FCS和SMD不同的中和效力与HIV-1包膜三聚体的结合。成功完成目标将在单个分子水平上提供新的定量方法，以理解病毒进入的早期步骤。该计划将提供一个极好的环境，在该环境中，申请人（在密切指导下）可以深入参与将高级荧光光谱和成像方法应用于HIV研究。随着候选人朝着生物医学光谱和成像研究的独立性发展，指导将更加针对生物学问题提供全面的投资组合。人类免疫缺陷病毒1型（HIV-1）一直是密集研究的主题。大多数抗逆转录病毒药物在感染后步骤起作用，无法治愈HIV-1感染。因此，一直着重于开发通过靶向HIV-1表面三聚体来阻止进入前事件的药物或疫苗，从而使HIV-1表面三聚体能够结合与细胞表面共感受器的结合。在此应用中，将在单分子水平上确定HIV-1包膜与细胞表面受体和/或抗Envelope抗体的性质。这种新的基于单分子荧光方法的应用预计将提供与疫苗和药物开发相关的独特信息。