title: Integration of chemical and RNAi multiparametric profiles identifies triggers of intracellular mycobacterial killing.
abstract: Pharmacological modulators of host-microbial interactions can in principle be identified using high-content screens. However, a severe limitation of this approach is the lack of insights into the mode of action of compounds selected during the primary screen. To overcome this problem, we developed a combined experimental and computational approach. We designed a quantitative multiparametric image-based assay to measure intracellular mycobacteria in primary human macrophages, screened a chemical library containing FDA-approved drugs, and validated three compounds for intracellular killing of M. tuberculosis. By integrating the multiparametric profiles of the chemicals with those of siRNAs from a genome-wide survey on endocytosis, we predicted and experimentally verified that two compounds modulate autophagy, whereas the third accelerates endosomal progression. Our findings demonstrate the value of integrating small molecules and genetic screens for identifying cellular mechanisms modulated by chemicals. Furthermore, selective pharmacological modulation of host trafficking pathways can be applied to intracellular pathogens beyond mycobacteria.
authors: Max-Planck Institute of Molecular Cell Biology and Genetics, 108 Pfotenhauerstrasse, 01307 Dresden, Germany.
authors: Varadharajan
authors: V
authors: Sundaramurthy
authors: Rico
authors: R
authors: Barsacchi
authors: Nikolay
authors: N
authors: Samusik
authors: Giovanni
authors: G
authors: Marsico
authors: Jerome
authors: J
authors: Gilleron
authors: Inna
authors: I
authors: Kalaidzidis
authors: Felix
authors: F
authors: Meyenhofer
authors: Marc
authors: M
authors: Bickle
authors: Yannis
authors: Y
authors: Kalaidzidis
authors: Marino
authors: M
authors: Zerial
mesh_terms: D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000494:pharmacology*; D001343:Autophagy / Q000187:drug effects*; D001692:Biological Transport; D015169:Colony Count, Microbial; D019295:Computational Biology / Q000379:methods; D004705:Endocytosis; D011992:Endosomes; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006220:Haloperidol / Q000737:chemistry / Q000494:pharmacology; D006367:HeLa Cells; D054884:Host-Pathogen Interactions / Q000187:drug effects; D006801:Humans; D008264:Macrophages / Q000187:drug effects / Q000382:microbiology / Q000648:ultrastructure; D008826:Microbial Sensitivity Tests; D050296:Microbial Viability / Q000187:drug effects; D009169:Mycobacterium tuberculosis / Q000187:drug effects* / Q000472:pathogenicity; D009661:Nortriptyline / Q000737:chemistry / Q000494:pharmacology; D010588:Phagosomes; D011346:Prochlorperazine / Q000737:chemistry / Q000494:pharmacology; D034622:RNA Interference
chemical_list: D000900:Anti-Bacterial Agents; D049452:Green Fluorescent Proteins; D009661:Nortriptyline; D006220:Haloperidol; D011346:Prochlorperazine