Quantitative Analysis of Trimethylamine Oxide and Related Metabolites

Trimetlylamine oxide (TMAO) is a small molecule compound widely found in aquatic products in nature, also exists in mammals, plants and fungi. It can participate in many important biological functions of the body, such as osmotic pressure regulation , maintain cellular homeostasis, etc. In recent years, with the depth study of intestinal flora, scientists have found that the levels of TMAO and related metabolites are closely related to cardiovascular disease, myocardial infarction, stroke, diabetes, chronic kidney disease, cancer and other diseases. Therefore, TMAO has received more and more attention, and related studies have been reported.

---

---

Sample Requirements

• Animal tissue ≥ 0.2g
• Stool ≥ 0.3g
• Serum, plasma≥ 0.3ml
• Cells ≥ 5×10⁶
• Store in liquid nitrogen or -80°C
• Dry ice shipping

---

Case Analysis

Development of a Gut Microbe-targeted Nonlethal Therapeutic to Inhibit Thrombosis Potential

Journal: Nature medicine       Impact factor: 36.13 Published date: September, 2018       Published by: Cleveland Lerner Institute, United States

Research Background

Trimethylamine N-oxide (TMAO) is a gut microbiota–derived metabolite that enhances both platelet responsiveness and in vivo thrombosis potential in animal models, and TMAO plasma levels predict incident atherothrombotic event risks in human clinical studies. TMAO is formed by gut microbe–dependent metabolism of trimethylamine (TMA) moiety-containing nutrients, which are abundant in a Western diet.

Research Method

TMA-generating enzyme pair, CutC and CutD (CutC/D), we developed inhibitors that are potent, time-dependent, and irreversible and that do not affect commensal viability. In animal models, a single oral dose of a CutC/D inhibitor significantly reduced plasma TMAO levels for up to 3 d and rescued diet-induced enhanced platelet responsiveness and thrombus formation, without observable toxicity or increased bleeding risk. The inhibitor selectively accumulated within intestinal microbes to millimolar levels, a concentration over 1-million-fold higher than needed for a therapeutic effect.

Research Result

These studies reveal that mechanismbased inhibition of gut microbial TMA and TMAO production reduces thrombosis potential, a critical adverse complication in heart disease. They also offer a generalizable approach for the selective nonlethal targeting of gut microbial enzymes linked to host disease limiting systemic exposure of the inhibitor in the host.

DMB attenuates choline diet–enhanced platelet responsiveness and in vivo rate of thrombus formation

The platelet activity index of mice did not change, but TMAO level and aggregation reaction increased sharply. DMB could reverse TMAO level and ADP-dependent platelet aggregation in mice fed a cholinergic diet, however, TMAO levels and platelet aggregation were not reversed after direct injection of TMAO.

DMB could significantly prolong the time of blood spot formation, but DMB was not effective after TMAO direct injection.

Design and development of potent, mechanism-based, nonlethal microbial CutC/D inhibitors.

Development of two potent inhibitors (IMC and FMC) that do not affect the viability of symbiotic bacteria but permanently inactivate CutC/D, a key enzyme for TMA production in bacteria.

CutC/D inhibitor in choline-fed mice

Accumulates in intestinal commensal bacteria, almost inhibit the production of TMA and TMAO caused by high choline diet completely, reduce platelet aggregation and thrombosis, and no obvious toxic side effects were observed.

Inhibition of choline TMA lyase activity alters gut microbiota

IMC and FMC significantly increased the abundance of akermann bacteria in the gut of cholinergic mice

Conclusion

These studies suggest a new potential target for the treatment of subjects at risk for thrombotic complications and cardiovascular disease.

Reference

Roberts Adam B,Gu Xiaodong,Buffa Jennifer A et al. Development of a gut microbe-targeted nonlethal therapeutic to inhibit thrombosis potential.[J] .Nat. Med., 2018, 24: 1407-1417