DEVELOPING a Pan-Coronavirus Vaccine

With mentor Prof.Dr.Lbachir BenMohamed at the University of California Irvine. In picture are colleagues and fellow scientists Dr.Nisha Dhanushkodi, Dr.Ruchi Srivastava, and Dr.Pierre Gregoire Coulon.

Further, my research focused on accessing the molecular profiling of genes associated with herpes pathophysiology. Knowing herpes specific milieu of genes and major biological pathways and the existing knowledge of immunology will aid in the development and implementation of novel immunotherapeutic strategies. Working on various projects related to Vaccine Development against Herpes Simplex Virus (HSV) infection. Major research involvements are:

Ø  Examined the efficacy of a therapeutic vaccine against ocular HSV-1 infection using HLA-A2 Transgenic rabbits.

Ø  Studied the roles of PD-1 and LAG-3 blockade towards an increased frequency of functional HSV-specific memory CD8+ T cells among (i) symptomatic and asymptomatic humans, and (ii) HSV-1 infected “humanized” HLA- Transgenic rabbits. Identified how the transcription milieu changes in response to the T cell exhaustion molecules' blockade during herpes infection through high throughput single-cell RNA sequencing and bulk-RNA sequencing. Validated the functional relevance of prominent genes through Flow Cytometry, Immunohistochemistry, Western Blot, and Real-Time PCR. Accessed how the HSV-1 viral titer changes in response to PD-1 and LAG-3 specific blockade by performing ex-vivo culture on trigeminal ganglia explants.

Ø  Evaluated the immune evasion mechanisms by which HSV-1 and HSV-2 employ their acute or latent genes to interfere with the host immune system during acute and latent viral infections. Further, examined the role of HSV- 1 latency-associated transcript (LAT) using the mouse model through UV-B induced reactivation. Profiled the differentially expressed genes and micro-RNAs using NanoString based gene and miRNA expression assays.

Ø  Identified epitopes/peptides from the HSV seropositive individuals by the genome-based bioinformatics searches and the prediction algorithms. Immunization of these peptides induced CD8+ T cell-dependent protective immunity against ocular herpes infection and disease in “humanized” HLA-A*02:01 transgenic mice. These findings should guide the development of a safe and effective T-cell-based herpes vaccine.

These projects involved conducting experiments, method development, optimization, and troubleshooting using different molecular/immunology/cell biology techniques, data analysis and presenting data in conferences and group meetings. I have already published over 10 research articles being part of this research group since May 2018, with two lead authorship papers.

Prakash S, Roy S, Srivastava R, Coulon PG, Dhanushkodi NR, Vahed H, Jankeel A, Geertsema R, Amezquita C, Nguyen L, Messaoudi I, Burkhardt AM, BenMohamed L. Unique molecular signatures of antiviral memory CD8+ T cells associated with asymptomatic recurrent ocular herpes. Sci Rep. 2020 Aug 14;10(1):13843. doi: 10.1038/s41598-020-70673-z. PMCID: PMC7427992.

While working at the Sanjay Gandhi Postgraduate Institute of Medical Sceinces, Lucknow; I was part of two different projects which (i) Evaluated the impact of miRNAs on ESRD and renal allograft dysfunction and (ii) Evaluated the clinical and environmental factors associated with Chronic Kidney Disease of Unknown Etiology (CKDu) by the Indian Council of Medical Research and Indian Society of Nephrology. Major research outcomes were:

Ø  Putative association of microRNAs with corresponding target genes and SNPs in altered renal pathophysiology: Post-transcriptional events account for nearly 97% of kidney function involving the activities of miRNA like ncRNAs. miRNA being a negative regulator may suppress or up-regulate the target gene expression. Our findings observed this, in turn, affects the vascularization process resulting in end-stage renal disease (ESRD). After exhaustive in-silico analysis, I figured out the association of 20 most significant miRNAs and their corresponding target genes with ESRD. The Real-Time PCR based genotyping revealed synergistic relation between mutant genotypes of miRNA198, let7e, miRNA296, and their corresponding target genes FGFR1, STAT3, and TGFβ1 towards susceptibility to ESRD. Further, through ELISA and western blotting, the functional relevance of the target genes were evaluated on serum and renal biopsy samples. [Funding: Indian Council of Medical Research]

DECIPHERING THE GENETICS AND PATHOLOGY ASSOCIATED WITH CHRONIC KIDNEY DISEASE OF UNKNOWN ETIOLOGY (CKDu)