(Provided by the applicant) Abstract: Through infection of CD4 positive (CD4+) T lymphocytes, the Human Immunodeficiency Virus type 1 (HIV-1) has claimed twenty-five million lives since its discovery in 1983. Although it has been well established that HIV-1 initiates a T cell infection by binding to CD4 and chemokine coreceptors on the cell surface, the early events in HIV-1 infection of CD4+ T cells remain poorly understood. A variety of different techniques have been used over the years to study mechanisms of HIV-1 entry. However, one technical limitation that is inherent in all these methods is the inability to track the fate of a single HIV-1 virion from the very beginning of viral entry to chromosomal integration. In all these experiments, a population of viruses and cells were incubated and measured. Because each entry event can lead to proviral DNA integration with a finite probability, it is therefore difficult to establish a causative link between entry pathway and productive infection. The goal of this project is to develop a set of nanoscopic and novel technologies that we can harness to define the pathway and interactions by which HIV-1 infects CD4+ T cells. We are developing a technique based on optical tweezers that can manipulate a single HIV-1 virion, deliver it to CD4+ T cell and thus allows us to determine the fate of CD4+ T cell upon entry by a single virion. This technique will allow us to unambiguously define the molecular mechanisms of HIV-1 infection. Furthermore, we propose to measure directly the interactions between a single HIV-1 virion and receptors in the context of a live T cell. Collectively, these studies will contribute to a definitive and quantitative understanding of early events in HIV infection, which may help therapeutic development that is aimed to block HIV-1 entry to CD4+ T cells. The techniques developed herein can be useful for studying cellular uptake of not only viruses but other molecules, macromolecular assemblies and nanoparticles, and are applicable to a wide-range of ligand-receptor interactions on the cell surface. Public Health Relevance: The early events in HIV infection of CD4+ T cells are poorly understood. This proposal aims to develop a set of nanoscopic and other novel techniques to study HIV infection of CD4+ T cells in real time, one virion at a time. If successful, the results from this study will establish for the first time a causative link between HIV entry pathway and the productive infection of CD4+ T cells, which will help therapeutic development that is aimed to block HIV-1 entry to CD4+ T cells.

Agency
National Institute of Health (NIH)
Institute
Office of The Director, National Institutes of Health (OD)
Type
NIH Director’s New Innovator Awards (DP2)
Project #
3DP2OD008693-01S1
Application #
8458458
Study Section
Special Emphasis Panel (ZGM1-NDIA-S (01))
Program Officer
Basavappa, Ravi
Project Start
2011-09-30
Project End
2016-06-30
Budget Start
2011-09-30
Budget End
2016-06-30
Support Year
1
Fiscal Year
2012
Total Cost
$122,870
Indirect Cost
$37,657
Name
University of Michigan Ann Arbor
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
DeSantis, Michael C; Cheng, Wei (2016) Label-free detection and manipulation of single biological nanoparticles. Wiley Interdiscip Rev Nanomed Nanobiotechnol 8:717-29
Cheng, Wei (2016) The Density Code for the Development of a Vaccine? J Pharm Sci 105:3223-3232
Pang, Yuanjie; Song, Hanna; Cheng, Wei (2016) Using optical trap to measure the refractive index of a single animal virus in culture fluid with high precision. Biomed Opt Express 7:1672-89
DeSantis, Michael C; Kim, Jin H; Song, Hanna et al. (2016) Quantitative Correlation between Infectivity and Gp120 Density on HIV-1 Virions Revealed by Optical Trapping Virometry. J Biol Chem 291:13088-97
Hou, Ximiao; DeSantis, Michael C; Tian, Chunjuan et al. (2016) Optical manipulation of a single human virus for study of viral-cell interactions. Proc SPIE Int Soc Opt Eng 9922:
Cheng, Wei (2015) Mechanisms of HCV NS3 helicase monitored by optical tweezers. Methods Mol Biol 1259:229-55
Pang, Yuanjie; Song, Hanna; Kim, Jin H et al. (2014) Optical trapping of individual human immunodeficiency viruses in culture fluid reveals heterogeneity with single-molecule resolution. Nat Nanotechnol 9:624-30
Koh, Byumseok; Cheng, Wei (2014) Mechanisms of carbon nanotube aggregation and the reversion of carbon nanotube aggregates in aqueous medium. Langmuir 30:10899-909
Kim, Jin H; Song, Hanna; Austin, Jamie L et al. (2013) Optimized Infectivity of the Cell-Free Single-Cycle Human Immunodeficiency Viruses Type 1 (HIV-1) and Its Restriction by Host Cells. PLoS One 8:e67170
Arunajadai, Srikesh G; Cheng, Wei (2013) Step detection in single-molecule real time trajectories embedded in correlated noise. PLoS One 8:e59279

Showing the most recent 10 out of 12 publications