Aryan
Sharma
Sponsor: Pamela Lein, Ph.D. VM: Molecular Bio Sciences Acute intoxication with organophosphate (OP) cholinesterase inhibitors, which include pesticides and nerve agents, can trigger life-threatening seizures. While current standard-of-care (SOC) increases survival rate, it does not protect against the development of acquired epilepsy and cognitive decline. Therefore, there is an urgent need for more effective therapeutic strategies. We are investigating TGF-β as a potential therapeutic target using a rat model of acute intoxication with the OP diisopropylfluorophosphate (DFP). Adult male and female rats were administered DFP or vehicle (VEH, phosphate-buffered saline), followed by SOC. At 1, 3, 7, and 14 days post-exposure (DPE), rats were euthanized, and levels of TGF-β were quantified in brain tissue by quantitative PCR (qPCR) and western blotting. qPCR revealed a fourfold increase in TGF-β mRNA at 1 DPE and a fivefold increase at 7 DPE in the hippocampus and cortex. Western blot analysis detected total SMAD2/3 protein, a downstream signaling molecule activated by TGF-β, in all samples, but revealed increased levels of phosphorylated SMAD2/3 (pSMAD2/3) in brains of DFP rats compared to VEH controls. Our findings indicate that TGF-β signaling is triggered by acute OP-exposure, supporting the validity of investigating TGF-β as a therapeutic target. This work was supported by NIH (U54 NS127758). Dimerization of Protein Kinase D1 in Vivo: Investigating the Role of the ULD Domain
Abstract profile. Full document pending author claim.
Authors:
Aryan Sharma
Date Created:
Not specified
Course Title:
Professor:
Not specified
About Paper:
Protein Kinase D1 (PKD1) is a serine/threonine kinase involved in cell survival and differentiation, protein trafficking, metabolism and gene transcription, making it highly relevant to diseases like heart failure and cancer. Despite its importance, the mechanisms regulating PKD1 activity remain unclear, particularly its ability to form dimers. Here, we investigate PKD1 dimerization in vivo, focusing on the Ubiquitin-like Domain (ULD), which may play a critical role in dimer formation and regulation. Using a fluorescence lifetime imaging microscopy (FLIM) approach, we generated donor and acceptor Förster resonance energy transfer (FRET) PKD1 fluorescent constructs to examine when, where, and how dimers form in a cellular context. We aim to determine whether PKD1 dimerizes at specific subcellular locations (membrane, nucleus, Golgi) and whether dimerization is constitutive or dynamically regulated. Additionally, we explore whether dimer populations can be shifted by kinase activation state and how ULD domain mutations impact dimer stability. Our findings will provide crucial insights into PKD1 regulation, offering potential therapeutic targets for PKD1-driven diseases. Effects of Social Media use on Adolescents' Mental Health Ashlyn Sharma
Source:
UC Davis / MED: Pharmacology / 2025
Topics:
No topics listed
Co-authors:
Aryan Sharma