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High-throughput analysis identifying drugs that reduce oxidative and ER stress in human coronary artery endothelial
Michael J Haas1, Victoria Feng1, Krista Gonzales1
1Division of Endocrinology, Diabetes, and Metabolism, Department of Medicine, University of Florida Jacksonville College of Medicine, Jacksonville, FL, USA.
Abstract:
Endoplasmic reticulum (ER) stress as well as oxidative stress have been shown to play important roles in metabolic and cardiovascular disease, and drugs that counteract the effects of ER and oxidative stresses may be clinically useful. To identify novel compounds that ameliorate ER and oxidative stresses, we screened two drug libraries purchased from Evotec, San Francisco, CA; the NIH clinical collection 1 (446 compounds) and the NIH clinical collection 2 (281 compounds). Human coronary artery endothelial cells (HCAEC) were tested for ER and oxidative stress. ER stress was measured with an ER stress-sensitive secreted alkaline phosphatase (SAP) assay. The cells were transfected with the plasmid pSAP2.Control, expressing a heat-resistant form of SAP, and treated with the ER stress inducer tunicamycin in the presence or absence of each of the various compounds for 24-h, at which time SAP activity was measured. Compounds exhibiting significant increases in SAP activity (41 compounds out of a total of 727 tested; 5.6%) were then assayed for their ability to suppress superoxide (SO) anion generation in cells treated with 27.5 mM dextrose. SO generation was measured using the superoxide-reactive probe 2-methyl-6-(4-methoxyphenyl)-3,7-dihydroimidazo[1,2-A]pyrazin-3-one hydrochloride chemiluminescence. Of the 41 compounds identified as ER stress reducers, only 33 (80.5%) suppressed dextrose-induced SO anion generation. Interestingly, 51% of the compounds found to be dual-stress modifiers consisted of cardioprotective drugs, including statins, angiotensin receptor blockers, angiotensin-converting enzyme inhibitors as well as β-blockers. Future studies to validate the clinical effectiveness of these agents remain to be performed in pre-clinical and clinical trials.
Insights
Researchers screened drug libraries to find compounds reducing endoplasmic reticulum (ER) and oxidative stress, key factors in cardiovascular disease. Many identified dual-action compounds were existing cardioprotective drugs, suggesting potential clinical applications.
Area of Science:
- Biomedical Sciences
- Pharmacology
- Cardiovascular Research
Background:
- Endoplasmic reticulum (ER) stress and oxidative stress are implicated in metabolic and cardiovascular diseases.
- Therapeutic agents mitigating these stresses could offer clinical benefits for related conditions.
Purpose of the Study:
- To identify novel compounds capable of ameliorating both ER and oxidative stress.
- To evaluate existing drug libraries for dual-action stress-modifying properties.
Main Methods:
- Screened 727 compounds from two NIH clinical collections using human coronary artery endothelial cells (HCAEC).
- Measured ER stress via a secreted alkaline phosphatase (SAP) assay after tunicamycin induction.
- Assessed oxidative stress by measuring superoxide anion generation using a chemiluminescence probe in dextrose-treated cells.
Main Results:
- Identified 41 compounds (5.6%) that reduced ER stress.
- Of these, 33 compounds (80.5%) also suppressed oxidative stress.
- Notably, 51% of dual-action compounds were known cardioprotective drugs (statins, ARBs, ACE inhibitors, beta-blockers).
Conclusions:
- Drug libraries contain compounds that can simultaneously reduce ER and oxidative stress.
- Existing cardioprotective medications exhibit dual-stress modifying effects, highlighting their potential therapeutic value.
- Further pre-clinical and clinical trials are warranted to validate these findings.
