Next-Generation Phenotypic Screening in Early Drug Discovery for Infectious Diseases.

Cell-based phenotypic screening has proven to be valuable, notably in recapitulating relevant biological conditions, for example, the host cell/pathogen niche. However, the corresponding methodological complexity is not readily compatible with high-throughput pipelines, and fails to inform either molecular target or mechanism of action, which frustrates conventional drug-discovery roadmaps. We review the state-of-the-art and emerging technologies that suggest new strategies for harnessing value from the complexity of phenotypic screening and augmenting powerful utility for translational drug discovery. Advances in cellular, molecular, and bioinformatics technologies are converging at a cutting edge where the complexity of phenotypic screening may no longer be considered a hinderance but rather a catalyst to chemotherapeutic discovery for infectious diseases.

Notch signaling in the pigmented epithelium of the anterior eye segment promotes ciliary body development at the expense of iris formation.

The ciliary body and iris are pigmented epithelial structures in the anterior eye segment that function to maintain correct intra-ocular pressure and regulate exposure of the internal eye structures to light, respectively. The cellular and molecular factors that mediate the development of the ciliary body and iris from the ocular pigmented epithelium remain to be fully elucidated. Here, we have investigated the role of Notch signaling during the development of the anterior pigmented epithelium by using genetic loss- and gain-of-function approaches. Loss of canonical Notch signaling results in normal iris development but absence of the ciliary body. This causes progressive hypotony and over time leads to phthisis bulbi, a condition characterized by shrinkage of the eye and loss of structure/function. Conversely, Notch gain-of-function results in aniridia and profound ciliary body hyperplasia, which causes ocular hypertension and glaucoma-like disease. Collectively, these data indicate that Notch signaling promotes ciliary body development at the expense of iris formation and reveals novel animal models of human ocular pathologies.

Manipulation of cells using an ultrasonic pressure field.

A novel device is described that generates an ultrasonic force field in a fluid layer. The force field arises because of the acoustic radiation force, a second order effect, generated as an ultrasonic wave interacts with a suspended particle. This force field can be used to manipulate objects in the fluid layer trapped between this device and an arbitrary surface, in this case, a flat object slide. The device is shown to be capable of positioning and, in doing so, concentrating human cells to predictable locations. Mesenchymal and HeLa cells were used. Critically, the forces required to do this can be generated by ultrasonic pressure fields that do not affect the viability of the cells. The viability has been assessed using trypan blue dye. The device used consists of a 14 mm square glass plate that is excited by at least one of four piezotransducers attached to the edges. The resulting ultrasonic force field and, importantly, the location of the minima in the force potential at which the cells are collected, has been calculated analytically.

High-throughput cell manipulation using ultrasound fields.

A method is presented for high-throughput cell manipulation that is capable of trapping and transporting biological cells in liquid using acoustic forces in an ultrasound field. The authors applied this technique for concentrating several cell types such as HeLa cells and human mesencymal stem cells. More than 90% of the cells were successfully concentrated into desired patterns. They also investigated cell viability in ultrasound fields and found little adverse effect. This work demonstrates that ultrasonic cell manipulation is suitable for being integrated into lab-on-a-chip systems for trapping and transporting large numbers of cells rapidly and is promising in cell fractioning.

High transfection efficiency of poly(4-vinylimidazole) as a new gene carrier.

Poly(4-vinylimidazole) (P4V) was obtained by free radical polymerization of 4-vinylimidazole (4V) prepared by decarboxylation of urocanic acid. P4V formed a complex with DNA that exhibited higher transfection effiency on Hela cells than polyethylenimine (PEI), through the proton sponge mechanism of the imidazole groups in the side chain of the P4V, and low cell toxicity.