BCP directly identifies the mode of action (MOA) of new candidates, information key to identifying molecules hitting new targets and for advancing these hits through the drug development pipeline. BCP reports MOA information for 100% of the hits obtained from screening, allowing investigators to immediately focus on the best hits in their collection.
BCP can detect cytological changes that do not kill or even impair growth, making it highly sensitive and allowing the discovery of molecules previously missed by other whole cell-based screens. The technology can also identify compounds that alter the cell in critical ways useful for developing new treatments. For example, BCP can identify compounds that permeabilize the outer membrane of Gram negative bacteria without affecting the inner membrane or cell growth. Such permeabilization agents sensitize bacteria to many clinically approved compounds and can be used in combination therapy to enhance susceptibility of Gram negative pathogens to many conventional antibiotics.
BCP works with all types of potential antimicrobial compounds, including small synthetic chemicals, crude natural product extracts, fractionated natural products, peptides, or synthetic molecules that mimic peptides. BCP can identify molecules that have multiple MOAs which are traditionally more difficult to characterize but highly desired due to their inherently low rates of resistance. Similarly, BCP can distinguish different MOAs which may arise in a mixture of molecules or a natural product extract.
Linnaeus has also developed a high throughput version of BCP (HTP-BCP) for identifying hits from large chemical or natural product libraries with unparalleled sensitivity.
Fungal Cytological Profiling (FCP)
Linnaeus has leveraged its experience with bacterial systems to develop its Fungal Cytological Profiling (FCP) platform to help accelerate the discovery and development of novel anti-fungals, a badly needed class of anti-infectives. While cytological profiling was initially developed in eukaryotic systems, such as mammalian cell lines, and groups such as Gebre et. al. have shown that such profiling can provide insight into the mechanism of action of anti-fungal drugs in the yeast Saccharomyces cerevisiae, it has not previously been developed for drug discovery. Linnaeus, through its partnership with Amplyx Pharmaceuticals, Inc., has met that challenge and developed platforms for C. albicans, A.fumigatus and C.neoformans. This platforms allows Linnaeus to intervene early in the anti-fungal drug discovery pipeline to rapidly characterize MOA of candidate molecules, identify and prioritize leads and quickly provide rich SAR data.
C. albicans displays unique cytological profiles after 24 hours in response to drugs targeting different pathways.
Manogepix (4X MIC) has a dramatic effect on C. albicans over time.
Effect of Manogepix (APX001A) on the C. albicans Cell Wall.
Cells are stained with Calcofluor White (blue, chitin) and Concanavalin A (green, mannan/glucan).
Arylthiazole antibiotics targeting intracellular methicillin-resistant Staphylococcus aureus (MRSA) that interfere with bacterial cell wall synthesis
Eid I., Elsebaei M.M., Mohammad H., Hagras M., Peters C.E., Hegazy Y.A., Cooper B., Pogliano J., Pogliano K., Abulkhair H.S., Seleem M.N., Mayhoub A.S.
2017 European Journal of Medicinal Chemistry
Phenylthiazole Antibacterial Agents Targeting Cell Wall Synthesis Exhibit Potent Activity in Vitro and in Vivo against Vancomycin-Resistant Enterococci
Mohammad, H., Younis, W., Chen, L., Peters, C.E., Pogliano, J., Pogliano, K., Cooper, B., Zhang, J., Mayhoub, A., Oldfield, E., Cushman, M., Seleem, M.N.
2017 Journal of Medicinal Chemistry
Bacteriological profiling of diphenylureas as a novel class of antibiotics against methicillin-resistant Staphylococcus aureus
Mohammad, H., Younis, W., Ezzat, H.G., Peters, C.E., AbdelKhalek, A., Cooper, B., Pogliano, K., Pogliano, J., Mayhoub, A.S., Seleem, M.N.
2017 PLOS ONE
Rapid Inhibition Profiling in Bacillus subtilis to Identify the Mechanism of Action of New Antimicrobials
Lamsa, A., Lopez-Garrido, J., Quach, D., Riley, E. P., Pogliano, J., and Pogliano, K.
2016 ACS Chemical Biology
Bacterial Cytological Profiling (BCP) as a Rapid and Accurate Antimicrobial Susceptibility Testing Method for Staphylococcus aureus
Quach D.T., Sakoulas G., Nizet V., Pogliano J., Pogliano K.
Application of bacterial cytological profiling to crude natural product extracts reveals the antibacterial arsenal of Bacillus subtilis
Nonejuie, P., Trial, R. M., Newton, G. L., Lamsa, A., Ranmali Perera, V., Aguilar, J., Liu, W. T., Dorrestein, P. C., Pogliano, J., and Pogliano, K.
2016 Journal of Antibiotics
Azithromycin Synergizes with Cationic Antimicrobial Peptides to Exert Bactericidal and Therapeutic Activity Against Highly Multidrug-Resistant Gram-Negative Bacterial Pathogens
Lin L., Nonejuie P., Munguia J., Hollands A., Olson J., Dam Q., Kumaraswamy M., Rivera H. Jr, Corriden R., Rohde M., Hensler M.E., Burkart M.D., Pogliano J., Sakoulas G., Nizet V.
Small-Molecule Inhibitors of Gram-Negative Lipoprotein Trafficking Discovered by Phenotypic Screening
McLeod, S.M., Fleming, P.R., MacCormack, K., McLaughlin, R.E., Whiteaker, J.D., Narita, S., Mori, M., Tokuda, H., Miller, A.A.
2015 Journal of Bacteriology
Bacterial cytological profiling rapidly identifies the cellular pathways targeted by antibacterial molecules
Nonejuie P., Burkart M., Pogliano K., Pogliano J.
2013 Proceedings of the National Academy of Sciences (PNAS)
The Bacillus subtilis cannibalism toxin SDP collapses the proton motive force and induces autolysis
Lamsa, A., Lui, W.T., Dorrestein, P.C. , Pogliano, K.
2012 Molecular Microbiology