The Deconstructed Granuloma: A Complex High-Throughput Drug Screening Platform for the Discovery of Host-Directed Therapeutics Against Tuberculosis

Huang, Lu; Kushner, Nicole L.; Theriault, Monique E.; Pisu, Davide; Tan, Shumin; McNamara, Case W.; Petrassi, H. Mike; Russell, David G.; Brown, Alexander

doi:10.3389/fcimb.2018.00275

Cited by 24 publications

(28 citation statements)

References 27 publications

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“…The impetus for research on this topic is further exacerbated by the inappropriate use of rich bacterial broth in studies that seeks to mimic the physiology of the site of infection of Mtb , which on the contrary is nutrient limiting. This has in fact been one of the most cited reasons for the failure of drug discovery programs [ 121 ] and establishment of in vitro conditions representative of in vivo infection and dormancy is critical for TB drug discovery.…”

Section: Metabolism In Drug Discoverymentioning

confidence: 99%

Metabolic Versatility of Mycobacterium tuberculosis during Infection and Dormancy

Chang

Guan

2021

Metabolites

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Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), is a highly successful intracellular pathogen with the ability to withstand harsh conditions and reside long-term within its host. In the dormant and persistent states, the bacterium tunes its metabolism and is able to resist the actions of antibiotics. One of the main strategies Mtb adopts is through its metabolic versatility—it is able to cometabolize a variety of essential nutrients and direct these nutrients simultaneously to multiple metabolic pathways to facilitate the infection of the host. Mtb further undergo extensive remodeling of its metabolic pathways in response to stress and dormancy. In recent years, advancement in systems biology and its applications have contributed substantially to a more coherent view on the intricate metabolic networks of Mtb. With a more refined appreciation of the roles of metabolism in mycobacterial infection and drug resistance, and the success of drugs targeting metabolism, there is growing interest in further development of anti-TB therapies that target metabolism, including lipid metabolism and oxidative phosphorylation. Here, we will review current knowledge revolving around the versatility of Mtb in remodeling its metabolism during infection and dormancy, with a focus on central carbon metabolism and lipid metabolism.

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Section: Metabolism In Drug Discoverymentioning

confidence: 99%

Metabolic Versatility of Mycobacterium tuberculosis during Infection and Dormancy

Chang

Guan

2021

Metabolites

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“…The substantial physiological reprogramming that occurs within mycobacteria during infection can render mycobacteria susceptible to therapies in the host that have no effect under standard in vitro growth conditions 63 . There is a growing recognition of the importance of developing models that reflect the in vivo cellular composition and reprogramming observed during mycobacterial infections 64 . Thus, the combination of chemical throughput, physiological context and the diversity of host and bacterial reporters available within the Myco-GEM model will enable screening for novel therapeutics.…”

Section: Discussionmentioning

confidence: 99%

An explant technique for high-resolution imaging and manipulation of mycobacterial granulomas

et al. 2018

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A central and critical structure in tuberculosis, the mycobacterial granuloma consists of highly organized immune cells, including macrophages that drive granuloma formation through a characteristic epithelioid transformation. Difficulties in imaging within intact animals as well as the inherent caveats of in vitro assembly models have severely limited the study and experimental manipulation of mature granulomas. Here we describe a new ex vivo granuloma culture technique, wherein mature, fully organized granulomas are microdissected and maintained in three-dimensional culture. This approach, in which granulomas retain key bacterial and host characteristics, enables high-resolution microscopy of granuloma macrophage dynamics, including epithelioid macrophage motility and granuloma consolidation. Through mass spectrometry, we find active production of key phosphotidylinositol species identified previously in human granulomas. We describe a method to transfect isolated granulomas, enabling genetic manipulation. In addition, we provide proof-of-concept for host-directed small molecule screens, identifying PKC signaling as an important regulator of granuloma macrophage organization.

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“…Taken together, Mm -infected zebrafish larvae coupled with improved automated imaging acquisition systems provide a reproducible and efficient model for: 1) evaluation of drug safety and toxicity; 2) rapid-screening of anti-tuberculosis drug effectiveness; 3) identification of potential HDT approaches. A limitation of screening in this model is that Mm antibiotic susceptibility is not identical to that of Mtb , so there is some risk of missing active compounds, but this in vivo screen may pick up other compounds that are only active in the presence of the host ( Dalton et al., 2017 ; Huang et al., 2018 ).…”

Section: Zebrafish Are An Effective Platform For Anti-tuberculosis Comentioning

confidence: 99%

One Size Fits All? Not in In Vivo Modeling of Tuberculosis Chemotherapeutics

Yang

Wang

Wen

et al. 2021

Front. Cell. Infect. Microbiol.

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Tuberculosis (TB) remains a global health problem despite almost universal efforts to provide patients with highly effective chemotherapy, in part, because many infected individuals are not diagnosed and treated, others do not complete treatment, and a small proportion harbor Mycobacterium tuberculosis (Mtb) strains that have become resistant to drugs in the standard regimen. Development and approval of new drugs for TB have accelerated in the last 10 years, but more drugs are needed due to both Mtb’s development of resistance and the desire to shorten therapy to 4 months or less. The drug development process needs predictive animal models that recapitulate the complex pathology and bacterial burden distribution of human disease. The human host response to pulmonary infection with Mtb is granulomatous inflammation usually resulting in contained lesions and limited bacterial replication. In those who develop progressive or active disease, regions of necrosis and cavitation can develop leading to lasting lung damage and possible death. This review describes the major vertebrate animal models used in evaluating compound activity against Mtb and the disease presentation that develops. Each of the models, including the zebrafish, various mice, guinea pigs, rabbits, and non-human primates provides data on number of Mtb bacteria and pathology resolution. The models where individual lesions can be dissected from the tissue or sampled can also provide data on lesion-specific bacterial loads and lesion-specific drug concentrations. With the inclusion of medical imaging, a compound’s effect on resolution of pathology within individual lesions and animals can also be determined over time. Incorporation of measurement of drug exposure and drug distribution within animals and their tissues is important for choosing the best compounds to push toward the clinic and to the development of better regimens. We review the practical aspects of each model and the advantages and limitations of each in order to promote choosing a rational combination of them for a compound’s development.

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The Deconstructed Granuloma: A Complex High-Throughput Drug Screening Platform for the Discovery of Host-Directed Therapeutics Against Tuberculosis

Cited by 24 publications

References 27 publications

Metabolic Versatility of Mycobacterium tuberculosis during Infection and Dormancy

Metabolic Versatility of Mycobacterium tuberculosis during Infection and Dormancy

An explant technique for high-resolution imaging and manipulation of mycobacterial granulomas

One Size Fits All? Not in In Vivo Modeling of Tuberculosis Chemotherapeutics

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