Current Projects in the Gupta Laboratory
Discovery of Novel Therapeutics via High-throughput Screening
We have a strong interest in identifying novel agents as potential therapeutics using high throughput, high content screening methodologies. Towards that end, we have developed a number of novel cell-based assays and technologies in the laboratory and are using them to screen compound collections. We have recently identified compounds that allosterically modulate function of leukocytic integrins. We have also developed novel, phenotypic assays with kidney podocytes.
Figure Legend: HTS Assay.Example photomicrographs from a 384-well plate assay showing cells remaining adherent upon completion of the no-wash cell adhesion assay. Cells were transferred to wells coated with an increasing amount of Fg and incubated in buffer containing either Ca2+ and Mg2+ (1 mM of each) or 1 mM Mn2+ for 20 min at 37 °C. Nonadherent cells were allowed to detach by gentle inversion of the plates, and the adherent cells were fixed with 1.1% formaldehyde. After washing the plate with the automated plate washer and staining cells with DAPI, cells were imaged using an automated microscope. The amount of Fg used to coat the wells is indicated. Cell adhesion to the uncoated well surface is
shown (no block).
Modulating inflammation via Integrin Activation
Our laboratory is focused on leukocyte-specific beta2 integrin family. Beta2 integrins (CD11a/CD18, CD11b/CD18, CD11c/CD18 and CD11d/CD18) are key to the biological function of leukocytes and mediate leukocyte adhesion and migration to the sites of inflammation. We have identified a number of small molecules that modulate the ligand-binding function of beta2 integrins. Detailed in vitro and in vivo characterization of these compounds is currently underway. In addition to being useful as therapeutics, the newly discovered molecules also serve as novel chemical biology probes to study the effects of functional modulation of integrins in vivo and to study the mechanism of integrin activation. We have also developed novel cell-based assays for High Throughput Screening (HTS) of chemical libraries and are using these assays to identify novel compounds.
Figure Legend: 3D ribbon model of integrin CD11b/CD18 (Mac-1, aMb2).
Novel Technologies for Molecular Diagnostics in Resource-Poor Settings
Another focus in the laboratory is towards development of novel assays and techniques for rapid, low-cost detection of infectious agents that are applicable in a low-resource setting. We have developed novel methods for DNA/RNA amplification that would allow for ready, multiplexed detection of multiple species using existing diagnostic systems and technologies and are currently optimizing these assays.
Protein-Protein interaction Networks and Signaling Pathways
In collaboration with Professor Mitsu Ogihara in the Department of Computer Science at University of Miami and with Professor M. J. Zaki, Professor of Computer Science and Bioinformatics at RPI, we are performing extensive modeling of a number of proteomics datasets (including some generated in our lab) to determine protein-protein interaction (PPI) networks in a variety of human cells and to devise new computational approaches for determining biologically relevant modules, dynamic changes in the PPI networks upon changes in cellular state and specific signaling pathways and their components.
Novel DNA-based nanomoters.
In this project area we are investigating applications of unique DNA and RNA sequences for producing novel structures (some self-assembling) and assemblies. We are also attempting to create DNA-templated nanoassembly and other nanomotors. Additionally, we are attempting to generate novel biologicals using the principles of Synthetic Biology.
Figure Legend: Three-dimensional model of a single-stranded DNA circle.