Molecular Pharmacology

Sympathetic neurons have a wide range of physiological functions and their hypoactivity contributes to obesity and diabetes, among other syndromes. Sympathomimemic drugs rescue this deficiency but this drug class, mostly composed of brain-penetrant amphetamines and adrenergic agonists, is both cardiotoxic and highly controlled. Our recent publication puts forward new class of drugs  named Sympathofacilitators that do not enter the brain and have an anti-obesity and cardio-neutral effect in vivo. The first-in-class was published in Mahu I et al Domingos, Cell Metabolism 2020; Fig. 3C of this paper demonstrated a neuro-facilitatory effect, rather than neuro-excitatory one. 

This new class is in needed of novel chemical entities which can be screened in vitro on cultured iPSC-derived sympathetic neurones. The screen would be based on fluorescent readouts of calcium activity reporter, screening for a facilitation of responses to acetylcholine (similar to Fig. 3C of Mahu I et al). 

The prospect of identifying natural compounds that have a Sympathofacilitatory effect is tangible when performed in collaboration with the laboratory of Barry O’Keefe. The student will learn lab how to grow and scale-up iPSC-derived sympathetic neurones in Domingos lab, and optimize an in vitro assay based on Fig. 3C. The student will then transfer this knowledge to the lab of Barry O’Keefe where the screen will be performed using a fluorescent plate reader, robotic liquid handling, and a library of natural compounds.

Our lab uses a variety of molecular, cellular and behavioural techniques to determine mechanisms of osteoarthritis pathogenesis and pain, using both mouse and naked mole-rat as model organisms. We are particularly interested in studying cell-cell interactions, for example identifying signalling pathways between fibroblast-like synoviocytes and knee-innervating neurons, using a combination of electrophysiology and behavioural assays. Past work has included the use of viral based modulation of neuronal function (i.e. chemogenetics), as well as exploring how mesenchymal stem cells modulate pain in osteoarthritis.

Sympathetic neurons have a wide range of physiological functions and their hypoactivity contributes to obesity and diabetes, among other syndromes. Sympathomimemic drugs rescue this deficiency but this drug class, mostly composed of brain-penetrant amphetamines and adrenergic agonists, is both cardiotoxic and highly controlled. Our recent publication puts forward new class of drugs  named Sympathofacilitators that do not enter the brain and have an anti-obesity and cardio-neutral effect in vivo. The first-in-class was published in Mahu I et al Domingos, Cell Metabolism 2020; Fig. 3C of this paper demonstrated a neuro-facilitatory effect, rather than neuro-excitatory one. 

This new class is in needed of novel chemical entities which can be screened in vitro on cultured iPSC-derived sympathetic neurones. The screen would be based on fluorescent readouts of calcium activity reporter, screening for a facilitation of responses to acetylcholine (similar to Fig. 3C of Mahu I et al).

The prospect of identifying natural compounds that have a Sympathofacilitatory effect is tangible when performed in collaboration with the laboratory of Barry O’Keefe. The student will learn lab how to grow and scale-up iPSC-derived sympathetic neurones in Domingos lab, and optimize an in vitro assay based on Fig. 3C. The student will then transfer this knowledge to the lab of Barry O’Keefe where the screen will be performed using a fluorescent plate reader, robotic liquid handling, and a library of natural compounds.

Sympathetic neurons have a wide range of physiological functions and their hypoactivity contributes to obesity and diabetes, among other syndromes. Sympathomimemic drugs rescue this deficiency but this drug class, mostly composed of brain-penetrant amphetamines and adrenergic agonist, is both cardiotoxic and highly controlled. Our recent publication puts forward new class of drugs named Sympathofacilitators that do not enter the brain and have an anti-obesity and cardio-neutral effect in vivo. The first in-class was published in Mahu I, Domingos, et al. Cell Metabolism 2020; Figure 3C of this paper demonstrated a neuro-facilitatory effect, rather than neuro-excitatory one.

This new class is in need of novel chemical entities which can be screened in vitro on cultured iPSC-derived sympathetic neurons. The screen would be based on fluorescent readouts of calcium activity reporter, screening for a facilitation of responses to acetylcholine (similar to Figure 3C of Mahu I, et al).

The prospect of identifying natural compounds that have a Sympathofacilitatory effect is tangible when performed in collaboration with the laboratory of Barry O'Keefe. The student will learn how to grow and scale-up iPSC-derived sympathetic neurons in the Domingos lab, and optimize an in vitro assay based on Figure 3C. The student will then transfer this knowledge to the lab of Barry O'Keefe where the screen will be performed using a fluorescent plate reader, robotic liquid handling, and a library of natural compounds.

The opportunistic human pathogen, Pseudomonas aeruginosa, is a commonly-found inhabitant in the airways of patients with chronic respiratory ailments such as COPD and cystic fibrosis (CF). Short chain fatty acids (SCFAs) such as acetate and propionate accumulate to high levels in the airways of these patients. In mutants of P. aeruginosa that are unable to catabolise SCFAs, these compounds are toxic and lead to cessation of growth. In this project, we aim to use fragment-based drug discovery to identify inhibitors of the key enzymes involved in propionate catabolism (PrpB and PrpC) and acetate assimilation (AceA). We have recently solved the x-ray crystal structure of each enzyme, and are supported by the Diamond Light Source to initiate a FBDD programme. Challenges will be to identify high affinity binders with specificity for the intended targets. Cell permeability and efflux of the “hits” will need to be investigated, as will “off target” effects, cytotoxicity to mammalian cells, and likely resistance mechanisms. Species specificity of inhibition will be examined in an in vitro polymicrobial system recently developed in the lab.