Biomarker Discovery, Cell line Development, Assay development, Protein Production and purification.

My current research is focusing on the study of hypothalamic melanocortin activity using Cerebrospinal Fluid (CSF) as a surrogate in lean, obese and weight reduced human subjects and in rodent models. We aim to validate biomarkers that can be used to predict responses to a given weight loss medication and to characterize counter regulatory responses to drug therapy that may limit efficacy. • Designed and validated ELISA to measure beta-endorphin in human CSF samples.• Using in house developed ELISA, identified POMC as a biomarker for weight loss medication targeting the serotonin receptor.• In collaboration with Washington University, St Louis, established circadian rhythm for brain glucocorticoids using Cerebrospinal Fluid (CSF) measurements as surrogate (J Clin Endocrinol Metab, 2019).• Analyzed the effect of opioid receptor antagonist, Naltrexone, on neuropeptides in human CSF samples and mouse hypothalamus.• Discovered alpha-MSH deactivation averts the efficacy of Naltrexone mediated weight loss in rodent model (J Endocr Soc 2109).• Reported acute sleep deprivation does not affect the diurnal rhythm of brain neuropeptides and glucocorticoids. However, such disturbance causes an elevation of brain biomarker beta-amyloid (J Alzheimers Dis 2020).• Collaborated with Indiana University and discovered G-protein receptor 17 can be used as an anti-obesity target against diet induced obesity (Nutr Diabetes. 2019).• Discovered Plasma-AgRP as the downstream effector of growth hormone (GH) on energy metabolism in acromegaly (a rare disease characterized by excess growth hormone) patients (J Clin Endocrinol Metab, 2019).• Established the glucocorticoid receptor (GR) mediated AgRP regulation in Cushing’s disease (J Clin Endocrinol Metab, 2019).• Managed sample data base and inventory in all human study trials.• Trained medical students and research associates.

RAD9 functions as a member of 9-1-1 (RAD9-RAD1-HUS1) complex recruited to the DNA damage site by DNA Damage Response (DDR) proteins. My study on three major projects focused on RAD9 role on prostate carcinogenesis, base excision repair and radio resistance.• Discovered two new functional roles of RAD9; one as a transcription factor and other as a protein complex stabilizer in addition to its canonical role of DNA repair. • Showed evidence of RAD9 as a transcription factor in human prostate cancer by studying target genes like NDRG1, FOXP1, NEIL1 and AGR2 (Carcinogenesis, 2019)• Utilizing mouse embryonic stem cells shown that RAD9 binds to Neil1 and prevents the later from proteasomal degradation (Nucleic Acids Res, 2014)• Showed binding of RAD9 to the p53 consensus sequences of its target genes.• Extended RAD9’s role in tumor hypoxia and glucose metabolism in human prostate cancer cells as evidenced human prostate cancer cells knocked out for RAD9 by CRISPR/cas9 have low glucose uptake as well as lower lactate production compared to the parental ones.• Utilizing CRISPR/cas9 gene editing technology and several cancer cell line models discovered RAD9 as a biomarker for radio resistance and other DNA damaging agents (Rad Oncology, 2014).• Optimized new in vitro assay system to measure glycosylation activity of NEIl1.• Identified downstream effectors or RAD9 those influence prostate carcinogenesis utilizing gene editing, protein expression and in vitro cell bases assays.

Involved in various projects revealing the study of Cyclin A and E on spermatogenesis as well as male fertility. • Generating reporter deletion and mutation constructs, luciferase reporter assay, gel shift assay, Chromatin immunoprecipitation, characterized the cis and trans regulatory elements responsible for testis specific expression of cyclin A1 (PLoS one, 2012).• Showed evidence that testis specific expression of cyclin A1 is independent to DNA methylation by CpG methylation profiler assay in various tissues. • Utilizing germ cell separation, flow cytometry and immunoprecipitation, identified and cyclinA1 interacting proteins in spermatocytes and deciphered their role in spermatogenic development as well as apoptosis in arrested spermatocytes (PLoS One, 2017).• Identified the downstream effectors in the apoptotic pathway of ccna1 deficient spermatocytes.• Discovered E type cyclins (E1 &E2) as regulators of homologous pairing of sex chromosomes (PloS Genetics, 2014).• Trained graduate students, research associates as well as summer students.