An Innovative Mass Spectrometry-Based Workflow for Drug Discovery

Descripción del proyecto

Descubrimiento de fármacos para la hipertensión pulmonar

La hipertensión arterial pulmonar (HTP) es una enfermedad rara relacionada con la hipertensión arterial en los vasos sanguíneos que irrigan los pulmones. La mayoría de los fármacos abordan la vasoconstricción inducida provocada por el estrechamiento o la obstrucción de las arterias pequeñas en los pulmones. Pruebas recientes señalan que los inhibidores de la proteína cinasa asociada a Rho (ROCK, por sus siglas en inglés) pueden beneficiar a los enfermos con HTP al actuar de manera específica contra la remodelación vascular y la inflamación en los pulmones. El proyecto MS4Drug, financiado con fondos europeos, tiene por objeto caracterizar los cambios conformacionales en las ROCK «in vitro», así como en el entorno celular inducido por estos inhibidores. Sus investigadores estudiarán los mecanismos moleculares que subyacen a las rutas de señalización de las ROCK y, de este modo, esperan poder diseñar fármacos mejores para la HTP.

Objetivo

Pulmonary Arterial Hypertension (PAH) is a life-threatening condition; even if treated, patients only have a 5-year survival rate of less than 60%. For the treatment of PAH, Chiesi Pharmaceuticals develops Rho-associated, coiled-coil-containing protein kinase (ROCK) inhibitors.
The aim of this project is to elucidate conformational changes in ROCK in vitro as well as in the cellular environment that are induced by anti-PAH drug candidates, which are currently being tested in preclinical trials at Chiesi Pharmaceuticals. The ROCK interactome will be mapped to gain detailed insights into the molecular mechanisms underlying the signaling pathways of ROCK. We will decipher the effect of anti-PAH drug candidates on the ROCK interactome and elucidate drug-induced conformational changes in ROCK. Identifying novel pharmacological targets modulating specific ROCK/protein interactions will eventually lead to novel drugs for an improved treatment of PAH.
This project will integrate, for the first time, in cell cross-lining mass spectrometry (XLMS), in conjunction with hydrogen/deuterium exchange mass spectrometry (HDX-MS), in the drug discovery pipeline of a pharma company. It will be the first application of the XLMS approach in the field of structural biology in Italy. The conformations and interactions of ROCK, derived by in cell XLMS, will allow a systematic validation of in vitro biochemical studies. This project aims to advance the in cell XLMS approach into a routine method for drug discovery on the system-wide level that complements HDX-MS studies using isolated proteins. Novel cross-linking reagent will be developed to bring the XLMS approach to a new level of time-resolved protein interaction studies. As such, the integrated approach described herein opens unmatched and novel perspectives for biotechnological and pharmaceutical companies.
The outcome of this project is expected to have a large impact on structural biology and drug discovery in general.