That news release about the DSMB was a breath of fresh air to break the long, quiet summer. But things are about to pick up. Several things to look forward to in the next 5 weeks.

1) European Society of Cardiology symposium in Rome August 27-August 31. A very impressive 1 hour session that Resverlogix is organizing titled: A novel approach for high CV risk patients & diabetes: the potential for epigenetics on August 28th dedicated entirely to novel approach of BET inhibition in cardiovascular disease. Big names like Kausik Ray, Jorge Plutzky, John Kastelein and Stefano Del Prato focusing on BET inhibition. Ewelina Kulikowski will have a poster there too on August 28 following this session. The talk titles, presenters and schedule can be found here.

2) Rodman and Renshaw on Sept 11-13. Resverlogix will present on Sept 12 or 13th and according to Clayton Paradis, Resverlogix will be webcasting.

3) European Association for the Study of Diabetes meeting in Munich Sept 12-16. They will present a poster entitled "Apabetalone (RVX-208) acts on epigenetics to lower Major Adverse Cardiovascular Events (MACE) in diabetes patients with atherosclerosis via microbiome activity," which focuses on a new mechanism of how RVX-208 beneficially affects the complement cascade via modulating the hepatic response of trimethylamine oxide (TMAO) to increase complement proteins, as well as RVX-208 suppressing levels of flavin-containing monooxygenase 3 (FMO3), the enzyme that makes TMAO. Full abstract at end of this message.

4) BioPharm America in Boston Sept 13-15. Resverlogix is listed as a presenting company

EASD abstract.

Apabetalone (RVX-208) acts on epigenetics to lower Major Adverse Cardiovascular Events (MACE) in diabetes patients with atherosclerosis via microbiome activity

N.C. Wong, C. Calosing, L. Tsujikawa, E. Kulikowski, D. Gilham, S. Wasiak, C. Halliday, J. Johansson, M. Sweeney

Background and aims: RVX-208 selectively inhibits the second ligand domain in bromodomain extra-terminal (BET) proteins that are epigenetic readers of acetylated lysine marks on histone tails. In the SUSTAIN and ASSURE (n=499) trials, patients (n=192) with diabetes mellitus (DM) and cardiovascular disease (CVD) given 200 mg/d of RVX-208 had a 77% relative risk reduction (RRR) in MACE. This marked RRR likely arises from RVX-208’s additive effects that include attenuation of known contributors to CVD such as the complement, inflammatory and coagulation pathways plus raising high density lipoproteins (HDL). In studies below, we tested a hypothesis that trimethylamine oxide (TMAO) a circulating metabolite connected to the microbiome may heighten complement activity. Next we asked how RVX-208 may beneficially affect the complement cascade (CC) in possibly underpinning the lower MACE observed in recent trials.

Materials and methods: Plasma from above human trials were analyzed. Gene expression data from RVX-208 exposed Huh-7 hepatoma cells and primary human hepatocytes.

Results: Heightened CC activity in CVD may perhaps be related to, a metabolite from dietary meat consumption, TMAO because of its ability to predict CVD risks. Human Huh-7 hepatoma cells (HC) grown in high glucose (25 mM) exposed to varying doses (0.1-100 uM) of TMAO revealed a dose and time dependent rise in mRNA encoding components of the CC; mannose-binding lectin (MBL) and C3 ranging from 1.2 to 1.5-fold. In high glucose and TMAO treated HC, adding 25 uM RVX-208 reduced levels of these mRNAs by >80%. Conversely, initial findings in HC grown exposed to euglycemia (5.6 mM glucose) TMAO failed to induce MBL or C3, but adding RVX-208 still suppressed both mRNAs by >80%. C5 another CC member was unlike MBL and C3 because TMAO did not induce C5 mRNA in HC exposed to hyperglycemia but RVX-208 also suppressed its expression. These findings point to the idea that TMAO enhances CC activity in hyperglycemia by inducing selected components of the CC but not all. In humans, plasma TMAO comes from flavin mono-oxygenase 3 (FMO3) activity on dietary components. To further explore the TMAO and CC link we had to use primary human hepatocytes (PHH) because FMO3 is not expressed in HC. Exposure of PHH to RVX-208 lowered FMO3 mRNA levels by 60%. Next we wondered whether these findings in vitro extended into humans by measuring CC proteins in plasma samples from our human trials. Consistent, in part with in vitro, MBL and C5 were significantly reduced by 15 and 12% respectively in plasma from patients given RVX-208 vs baseline in the ASSURE trial but C3 in the same samples did not change in a similar fashion.

Conclusion: HC grown in high glucose exposed to TMAO induces mRNAs encoding MBL and C3, two central components of the CC. Not only is TMAO induction of the mRNAs abrogated by apabetalone, it also suppresses by >80% levels of mRNAs encoding these CC. A separate effect of RVX-208 appears upstream of TMAO by suppressing FMO3 the enzyme that makes TMAO. Our findings suggest that RVX-208 has multiple ways of calming the CC at several points in liver cells exposed to hyperglycemia. Together our findings suggest that actions of RVX-208 may beneficially impact innate immunity by affecting CC in ways that could lead to lower MACE in patients with DM and CVD.