Real-time non-invasive fluorescence imaging of gut commensal bacteria to detect dynamic changes in the microbiome of live mice

Apostolos, Alexis J.; Chordia, Mahendra D.; Kolli, Sree H; Dalesandro, Brianna E.; Rutkowski, Melanie R.; Pires, Marcos M.

doi:10.1016/j.chembiol.2022.11.010

Cited by 11 publications

(10 citation statements)

References 47 publications

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“…In 2017, Hudak et al (and since then others − ) conjugated a fluorophore to a D -amino acid metabolic tag and found that this probe labeled bacteria in the gut of mice. At the same time, our laboratory showed in vivo metabolic labeling of peptidoglycan from Staphylococcus aureus harbored in Caenorhabditis elegans followed by a recent demonstration in mice …”

Section: Resultsmentioning

confidence: 96%

“…At the same time, our laboratory showed in vivo metabolic labeling of peptidoglycan from Staphylococcus aureus harbored in Caenorhabditis elegans 73 followed by a recent demonstration in mice. 74 To initially benchmark the metabolic labeling conditions, L. casei was cultured with DMSO or in the presence of D-KAz (Figure 6A). D-KAz bears an azido tag and it is expected to be metabolically incorporated throughout the peptidoglycan scaffold.…”

Section: ■ Resultsmentioning

confidence: 99%

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Noninvasive Analysis of Peptidoglycan from Living Animals

Ocius,

Kolli,

Ahmad

et al. 2024

Bioconjugate Chem.

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The role of the intestinal microbiota in host health is increasingly revealed in its contributions to disease states. The host-microbiome interaction is multifactorial and dynamic. One of the factors that has recently been strongly associated with host physiological responses is peptidoglycan from bacterial cell walls. Peptidoglycan from gut commensal bacteria activates peptidoglycan sensors in human cells, including the nucleotide-binding oligomerization domain-containing protein 2. When present in the gastrointestinal tract, both the polymeric form (sacculi) and depolymerized fragments can modulate host physiology, including checkpoint anticancer therapy efficacy, body temperature and appetite, and postnatal growth. To utilize this growing area of biology toward therapeutic prescriptions, it will be critical to directly analyze a key feature of the host-microbiome interaction from living hosts in a reproducible and noninvasive way. Here we show that metabolically labeled peptidoglycan/sacculi can be readily isolated from fecal samples collected from both mice and humans. Analysis of fecal samples provided a noninvasive route to probe the gut commensal community including the metabolic synchronicity with the host circadian clock. Together, these results pave the way for noninvasive diagnostic tools to interrogate the causal nature of peptidoglycan in host health and disease.

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Section: Resultsmentioning

confidence: 96%

Section: ■ Resultsmentioning

confidence: 99%

Noninvasive Analysis of Peptidoglycan from Living Animals

Ocius,

Kolli,

Ahmad

et al. 2024

Bioconjugate Chem.

Self Cite

View full text Add to dashboard Cite

show abstract

“…At the same time, our laboratory showed in vivo metabolic labeling of peptidoglycan in Caenorhabditis elegans 67 followed by a recent demonstration in mice. 68…”

Section: Resultsmentioning

confidence: 99%

Non-invasive Analysis of Peptidoglycan from Living Animals

Ocius

Kolli

Rutkowski

et al. 2023

Preprint

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The role of the intestinal microbiota in host health is increasingly revealed in its contributions to disease states. The host-microbiome interaction is multifactorial and dynamic. One of the factors that has recently been strongly associated with host physiological responses is peptidoglycan from bacterial cell walls. Peptidoglycan from gut commensal bacteria activate peptidoglycan sensors in human cells, including the Nucleotide-binding oligomerization domain containing protein 2 (NOD2). When present in the gastrointestinal tract, both the polymeric form (sacculi) and de-polymerized fragments can modulate host physiology including checkpoint anticancer therapy efficacy, body temperature and appetite, and postnatal growth. To leverage this growing area of biology towards therapeutic prescriptions, it will be critical to directly analyze sacculi from living hosts in a reproducible and non-invasive way. Here we show that peptidoglycan/sacculi can be readily isolated from fecal samples of mice and humans. Analysis of fecal samples provided a non-invasive route to probe the gut commensal community including the metabolic synchronicity with the host circadian clock. Together, these results pave the way for non-invasive diagnostic tools to emerge that inform on the causal nature of peptidoglycan in host health and disease.

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“…Further, we demonstrated that such analogs can be incorporated in vivo into the cell wall of S. aureus infected C. elegans [23h] and various diverse bacterial species residing in the gut microbiome of a mouse model. [25] To start, we prepared DBCO-tagged PGs by synthesizing the unnatural amino acid, D-diaminopropanoic acid (Dap), and conjugating its sidechain with DBCO to yield D-DapD. Incubation of this amino acid in the growth media of S. aureus results in the exchange of the terminal D-alanine on the PG stem peptide with the supplemented unnatural amino acid (Figure S1).…”

Section: Methodsmentioning

confidence: 99%

Measurement of Accumulation of Antibiotics to Staphylococcus aureus in Phagosomes of Live Macrophages**

Kelly,

Dalesandro,

Liu

et al. 2023

Angew Chem Int Ed

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Staphylococcus aureus (S. aureus) has evolved the ability to persist after uptake into host immune cells. This intracellular niche enables S. aureus to potentially escape host immune responses and survive the lethal actions of antibiotics. While the elevated tolerance of S. aureus to small‐molecule antibiotics is likely to be multifactorial, we pose that there may be contributions related to permeation of antibiotics into phagocytic vacuoles, which would require translocation across two mammalian bilayers. To empirically test this, we adapted our recently developed permeability assay to determine the accumulation of FDA‐approved antibiotics into phagocytic vacuoles of live macrophages. Bioorthogonal reactive handles were metabolically anchored within the surface of S. aureus, and complementary tags were chemically added to antibiotics. Following phagocytosis of tagged S. aureus cells, we were able to specifically analyze the arrival of antibiotics within the phagosomes of infected macrophages. Our findings enabled the determination of permeability differences between extra‐ and intracellular S. aureus, thus providing a roadmap to dissect the contribution of antibiotic permeability to intracellular pathogens.

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Real-time non-invasive fluorescence imaging of gut commensal bacteria to detect dynamic changes in the microbiome of live mice

Cited by 11 publications

References 47 publications

Noninvasive Analysis of Peptidoglycan from Living Animals

Noninvasive Analysis of Peptidoglycan from Living Animals

Non-invasive Analysis of Peptidoglycan from Living Animals

Measurement of Accumulation of Antibiotics to Staphylococcus aureus in Phagosomes of Live Macrophages**

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