Characterization of the Discrepancies Between Four-Dimensional Phase-Contrast Magnetic Resonance Imaging and In-Silico Simulations of Cerebrospinal Fluid Dynamics

The purpose of the present study was to compare subject-specific magnetic resonance imaging (MRI)-based computational fluid dynamics (CFD) simulations with time-resolved three-directional (3D) velocity-encoded phase-contrast MRI (4D PCMRI) measurements of the cerebrospinal fluid (CSF) velocity field in the cervical spinal subarachnoid space (SSS). Three-dimensional models of the cervical SSS were constructed based on MRI image segmentation and anatomical measurements for a healthy subject and patient with Chiari I malformation. CFD was used to simulate the CSF motion and compared to the 4D PCMRI measurements. Four-dimensional PCMRI measurements had much greater CSF velocities compared to CFD simulations (1.4 to 5.6 x greater). Four-dimensional PCMRI and CFD both showed anterior and anterolateral dominance of CSF velocities, although this flow feature was more pronounced in 4D PCMRI measurements compared to CFD. CSF flow jets were present near the nerve rootlets and denticulate ligaments (NRDL) in the CFD simulation. Flow jets were visible in the 4D PCMRI measurements, although they were not clearly attributable to nerve rootlets. Inclusion of spinal cord NRDL in the cervical SSS does not fully explain the differences between velocities obtained from 4D PCMRI measurements and CFD simulations.

An integrative in silico methodology for the identification of modulators of macrophage migration inhibitory factor (MIF) tautomerase activity

Farah El Turk, Bruno Claude Daniel Fauvet, Hilal Lashuel, Hajer Ouertatani-Sakouhi

Macrophage migration inhibitory factor (MIF) is a major proinflammatory cytokine that has been increasingly implicated in the pathogenesis of several inflammatory, autoimmune, infectious and oncogenic diseases. Accumulating evidence suggests that the tautomerase activity of MIF plays a role in modulating some of its intra- and extra-cellular activities. Therefore, the identification and development of small-molecule inhibitors targeting the catalytic activity of MIF has emerged as an attractive and viable therapeutic strategy to attenuate its function in health and disease. Herein we report a novel virtual screening protocol for the discovery of new inhibitors of MIF's tautomerase activity. Our protocol takes into account the flexibility and dynamics of the catalytic site by coupling molecular dynamics (MD) simulations aimed at modeling the protein's flexibility in solution to (i) docking with FlexX, or (ii) docking with FlexX and pharmacophoric filtering with Unity. In addition, we applied in parallel a standalone docking using the new version of Surflex software. The three approaches were used to screen the ChemBridge chemical library and the inhibitory activity of the top-ranked 333 compound obtained from each approach (1000 compound in total) was assessed in vitro using the tautomerase assay. This biochemical validation process resulted in the identification of 12 novel MIF inhibitors corresponding to a 1.2% hit rate. Six of these hits came from Surflex docking; two from FlexX docking with MD simulations and four hits were identified with MDS and pharmacophore filtering with minimal overlap between the hits from each approach. Six hits were identified with IC50 values lower than 10 microM (three hits with IC50 lower than 1 microM); four were shown to be suicide inhibitors and act via covalent modification of the N-terminal catalytic residues Pro1. One additional inhibitor, N-phenyl-N-1,3,4-thiadiazol-2-yl-thiourea, (IC50=300 nM) was obtained from FlexX docking combined to pharmacophoric filtering on one of the eight MD structures. These results demonstrate the power of integrative in silico approaches in the discovery of new modulator of MIF's tautomerase activity. The chemical diversity and mode of action of these compounds suggest that they could be used as molecular probes to elucidate the functions and biology of MIF and as lead candidates in drug developments of anti-MIF drugs.

Elsevier2010

An integrative in silico methodology for the identification of modulators of macrophage migration inhibitory factor (MIF) tautomerase activity

Farah El Turk, Bruno Claude Daniel Fauvet, Hilal Lashuel, Hajer Ouertatani-Sakouhi

Elsevier2010