AID	Name	Type	Comment
624149	qHTS of Nrf2 Activators: LOPAC Validation	confirmatory
624171	qHTS of Nrf2 Activators	confirmatory	qHTS AID
652013	qHTS of Nrf2 Activators: Hit Validation in NQO-1 Induction Assay	other
651595	qHTS of Nrf2 Activators: Hit Validation in Cytotox Assay	confirmatory
652014	qHTS of Nrf2 Activators: Hit Validation in GCLC Induction Assay	other
651593	qHTS of Nrf2 Activators: Hit Validation in Primary Assay	confirmatory
651597	qHTS of Nrf2 Activators: Hit Validation in Secondary FLuc Assay	confirmatory

Many diseases have some form of oxidative stress injury and ties to inflammation, causing a host of problems for the patient. The antioxidant response element (ARE) plays an important role in alleviating the harmful effects of oxidative stress. The antioxidant response element (ARE) is a transcriptional regulatory element involved in the activation of genes coding for a number of antioxidant proteins and detoxifying enzymes. These enzymes work in concert to protect tissues from oxidative insults and chemical toxicities in human hepatocytes and immune cells. The protective effects of ARE activation are primarily triggered through Nrf2 (NF-E2-related factor) binding to and activating AREs. It is known that Nrf2 controls the production of over 250 antioxidant and detoxification proteins (Hu et al., 2006), thereby protecting tissues by increasing cellular antioxidant content and suppressing inflammatory signaling pathways. The transcription factor Nrf2 is a central link between oxidative chemicals, such as phenolic antioxidants and electrophilic compounds, and the activation of ARE. Nrf2 levels are constitutively low as a consequence of its interaction with Keap1, which targets its degradation (Zipper and Mulcahy, 2002). Electrophiles react with key cysteine residues in Keap1, releasing Nrf2 and allowing its translocation to the nucleus. Once within the nucleus, Nrf2 complexes with coactivators such as p300, and binds to the AREs to induce gene transcription of cytoprotective enzymes, resulting in the prevention of toxicity. Activation of Nrf2 protects tissues by increasing cellular antioxidant content and suppressing inflammatory signaling pathways. Identifying novel and potent Nrf2 activators by using the AREc32 cell line to detect ARE activation by small molecules will lead to the identification and development of probes to study protective pathways in multiple tissues. These specific probes are essential to study the ARE pathway, and eventually to determine whether this pathway does activate genes that could protect against a host of diseases, including cardiovascular diseases, obesity, diabetes, Alzheimer's, and Parkinson's disease.

This project's aim is to identify novel and potent Nrf2 activators by using the AREc32 cell line to detect ARE activation by small molecules, which will lead to the identification and development of probes to study the ARE pathway.

NIH Chemical Genomics Center [NCGC]
NIH Molecular Libraries Probe Centers Network [MLPCN]

MLPCN Grant: DK081461
Assay Submitter (PI): Curtis Klaassen, University of Kansas Medical Center

Please see linked AIDs for detailed protocols.

This project is on-going and will be updated at a later point.

Grant Number: DK081461