Program Director/Principal Investigator: Santra, Santimukul
PROJECT SUMMARY/ABSTRACT
This application proposes to develop liposome-coated iron oxide nanosensors (LIONs) mimicking as
magnetically-labeled host membranes to detect fusion interactions of enveloped viruses. Spin-spin T2 relaxation
technique will be established as a generic bioanalytical method to detect dynamic conformational changes in
viral glycoproteins that play vital roles in fusion interactions. Within the first aim, influenza fusion protein
interactions with LIONs membrane will be studied, considering diverse fusion triggering environmental
factors including low pH, protease activation and receptor binding. Further studies to investigate the
sensitivity of this LIONs technology in quantifying similar fusion interactions with more native configurations of
HA is demonstrated using virus-like particle (VLPs) and shorter domains derived from HA. Similarly, fusion
experiments will be carried out using ganglioside-conjugated LIONs. The promising outcome of this research will
be screening potential antiviral candidates including small molecules and neutralizing antibodies.
The second aim establishes the broad adaptability of this sensitive LIONs technology for evaluation of fusion
interactions of enveloped viruses where presence of membrane receptor is an important trigger, for example,
SARS-CoV-2. Receptor-conjugated LIONs (R-LIONs) will be developed for the elucidation of this vital process
using spike proteins and reporter virus particles (RVPs) of SARS-CoV-2. R-LIONs offers a novel approach of
exploring the role of variety of entry receptors that may play an important role in SARS-CoV-2 membrane fusion.
The third aim of this proposal determines further advantage of nanosensor technology for the real-time
monitoring of virus-mediated cell fusion. SARS-CoV-2 virus entry into ACE2 receptor overexpressing mammalian
cells (HEK-293T) will be examined by developing magnetically labeled reporter virus particles (M-RVPs). Parallel
experiments will be carried out in the presence of fusion inhibitors and neutralizing antibodies. The important
application of this experiment will be demonstrated by screening potential new fusion inhibitors and antiviral
candidates.
The proposed AREA proposal, if successful, will develop novel nanosensor technology for the rapid detection of
fusion interactions of enveloped viruses including SARS-CoV-2 and influenza. This adaptable technology will
allow for rapid screening of potential drug candidates and fusion inhibitors of many other enveloped viruses, in
a timely fashion. Most importantly, this AREA proposal will initiate the proposed PURE program for
undergraduate education, and increase the hands-on biomedical research opportunity and learning for students
in our undergraduate-focused institution.
OMB No. 0925-0001 and 0925-0002 (Rev. 03/20 Approved Through 02/28/2023) Page Project Summary
