Domokos Gero

Title : Cell-based screening for anti-inflammatory drugs with the premise of innate immune tolerance induction

Abstract:

Inflammation is a poorly regulated reaction to pathogens or tissue injury that often requires medical therapy to prevent further tissue damage and to reduce the associated symptoms. Excessive inflammatory response may be triggered by evolutionarily conserved molecular determinants, termed pathogen- or damage-associated molecular patterns (PAMPs, DAMPs) that stimulate macrophages to produce inflammatory cytokines, including tumor necrosis factor-alpha (TNF-a), which in turn activates the inflammatory cascade. Current anti-inflammatory medications poorly block the early steps of inflammatory pathways and mainly act on downstream targets.

To find compounds that mimic innate immune tolerance and inhibit the initial steps of innate immune cell activation, we performed a cell-based screening campaign on a monocyte-macrophage cell-line. RAW264.7 cells were pre-exposed to biologically active drugs (Tocriscreen Plus Screening Library, 1280 compounds) and treated subsequently with the prototypical PAMP lipopolysaccharide (LPS). Inflammatory response was evaluated by measuring the TNF-a secretion with simultaneous assessment of cellular viability.

Test compounds induced a marked effect on cytokine production while hardly affected the cellular viability: 12.8% of drugs blocked the production of TNF-a by 50% or more. Our target-agnostic approach identified 20 clinical drugs that suppressed the inflammatory response (hit ratio of 1.5%) including steroids and the lipid-lowering atorvastatin, the b2-agonist salmeterol and the antidepressant escitalopram.

In conclusion, the initial steps of PAMP-induced inflammation may be modified by pharmacological interventions using currently available clinical drugs. Further testing of these compounds may be considered in experimental models of sepsis with a drug repurposing approach to test the translational potential of these findings.

¹Dr. Domokos Gero is supported by the Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences.

Audiencetake-away:

• A novel, target-agnostic approach to target inflammation
• A cell-based drug screening effort with a drug repurposing concept to target inflammation
• A summary of the results with approachable graphic representation of data giving an example of cell-based screen results.

Biography:

Dr. Domokos Gero graduated as an MD at the Semmelweis University Medical School in 2000 and also received his PhD degree at the Semmelweis University. In 2000, he started transgenic mouse research in Dr. Mozes’ laboratory focusing on gene therapeutic approaches against renal fibrosis at the Pathophysiology Department of Semmelweis University, then from 2006 he has started working on cell-based drug screening projects after he joined the CellScreen Research Center at the University. In 2010, he was invited to lead the cell-based drug screening group in Dr. Csaba Szabo’s lab and became a faculty of the Department of Anesthesiology, University of Texas Medical Branch (UTMB), USA and continued his work there on early drug discovery projects. In 2015 he moved to the UK, receiving a Marie Curie fellowship and joined the University of Exeter Medical School, Devon, UK for 2 years working with Dr. Matt Whiteman on further characterization of mitochondrial hydrogen-sulfide donors. Since 2017, he has been teaching at the Institute of Translational Medicine, Semmelweis University Medical School, Hungary and stated his own cell-based screening laboratory. He has published over 50 research papers.