Emanuel Holm, Isabeau Vermeulen, Saba Parween, Ana Lpez-Prez, Berta Cillero-Pastor, Michiel Vandenbosch, Silvia Remeseiro, Andreas Hrnblad
8 min
Metabolic dysfunction-associated steatotic liver disease (MASLD) is a major global health challenge with limited therapeutic options. Given that AMPK acts as a master regulator of cellular energy metabolism, researchers investigated whether ATX-304, a direct AMPK activator, could reverse the progression of fatty liver disease and associated metabolic dysfunction in mice.
Researchers used a mouse model fed a choline-deficient high-fat diet (CD-HFD) to induce progressive liver disease, including steatosis and fibrosis. Mice were treated with ATX-304 over short-term and long-term periods. The team employed a comprehensive analytical toolkit, including transcriptomics (RNA-seq), proteomics, spatial lipidomics (MALDI-MSI), and histology, to evaluate changes in liver function, lipid composition, and metabolic gene expression across different liver lobes.
ATX-304 treatment significantly improved the metabolic profile of the mice, notably reducing whole-body fat mass and lowering blood cholesterol levels. Within the liver, the drug induced a systemic 'catabolic switch,' characterized by suppressed lipid synthesis and enhanced fatty acid oxidation. This shift effectively reduced liver steatosis and limited the development of fibrosis. Interestingly, while the treatment was globally beneficial, the researchers noted pronounced lobular heterogeneities—where the left liver lobe often showed a more robust response than the right median lobe—and a distinct shift in the zonal distribution of lipid droplets within the liver lobules.
These results suggest that ATX-304 is a highly promising candidate for the clinical treatment of MASLD. By demonstrating that an AMPK activator can simultaneously manage whole-body adiposity, cholesterol, and specific hepatic pathology, the study provides a strong rationale for moving toward clinical trials. Furthermore, the findings highlight important spatial variations in how the liver responds to metabolic therapies, suggesting that future clinical assessments may need to account for localized differences in liver tissue health.
Metabolic dysfunction-associated steatotic liver disease (MASLD) is the most common chronic liver disease worldwide for which there is only one approved treatment. Adenosine monophosphate-activated protein kinase (AMPK) is an interesting therapeutic target since it acts as a central regulator of cellular metabolism. Despite efforts to target AMPK, no direct activators have yet been approved for treatment of this disease. This study investigated the effect of the AMPK activator ATX-304 in a preclinical mouse model of progressive fatty liver disease. The data demonstrated that ATX-304 diminishes body fat mass, lowers blood cholesterol levels, and mitigates general liver steatosis and the development of liver fibrosis, but with pronounced local heterogeneities. The beneficial effects of ATX-304 treatment were accompanied by a shift in the liver metabolic program, including increased fatty acid oxidation, reduced lipid synthesis, as well as remodeling of cholesterol and lipid transport. We also observed variations in lipid distribution among liver lobes in response to ATX-304, and a shift in the zonal distribution of lipid droplets upon treatment. Taken together, our data suggested that ATX-304 holds promise as a potential treatment for MASLD.
Alex: They stayed on it, with the drug mixed into the food. The comparisons were untreated mice on the same disease-inducing diet and mice eating regular chow. Different cohorts received treatment for shorter or longer periods.
Sam: Before we reach the liver, what happened to the animals as a whole?
Alex: Body weight fell rapidly after treatment began in the longer-term experiments. Body-composition measurements attributed that loss to fat rather than a reduction in lean mass. But treated animals gained slightly less lean mass over time than the comparison groups.
Sam: A drug mixed into food could make mice eat less. Could reduced intake account for the fat loss?
Alex: It could explain part of the initial loss in the longer-term cohort, because food intake initially declined. After that, treated animals ate more than untreated animals while maintaining low fat mass. That supports a metabolic change, although increased energy expenditure remained an interpretation rather than a direct measurement here.
Sam: Let’s separate the main liver result from its regional exception. How much did liver fat actually change?
Alex: After ten weeks of treatment, liver triglycerides—the main stored fats—were approximately sixty percent lower than in untreated mice on the disease-inducing diet. That comparison used ten animals per group. Ultrasound also suggested that treatment suppressed further progression of fatty liver.
Sam: Those measurements average across tissue. What changed when the researchers looked at individual liver regions?
Alex: Fat staining showed a strong response in the left lobe, but not the same improvement in the right median lobe. Scarring also fell in the left lobe, reaching levels seen in healthy-diet controls. In the right median lobe, however, the fibrosis score exceeded both untreated disease-model mice and healthy controls.
Sam: So calling the treatment antifibrotic without that qualification would hide a result going in the opposite direction.
Alex: It would. The authors describe improvement across the majority of liver volume, but the regional exception is central to interpreting it. The scarring analysis used five animals per group, so I’d also be cautious about how precisely these regional effects are estimated.
Sam: What supports the proposed explanation that the liver shifted from making fat toward burning it?
Alex: The researchers measured gene activity and protein abundance. Both pointed toward reduced fatty acid synthesis and increased beta-oxidation, a pathway that breaks down fatty acids. Several fat-transport proteins also increased, suggesting that treated livers could take up more fat from the circulation.
Sam: Increased uptake sounds potentially troublesome in a fatty liver. How do the authors reconcile that with less stored fat?
Alex: Their interpretation is that uptake, breakdown, and storage must be considered together. The liver could receive more fatty acids yet retain less if it breaks them down faster. Some genes involved in making stored triglycerides also increased, so this was not simply a shutdown of every fat-building process.
Sam: Do those molecular results explain why one lobe improved more than another?
Alex: Not convincingly. Gene and protein profiles showed broadly similar metabolic programs across the treated lobes. In the longer-term gene analysis, very few genes differed significantly in the direct treated-versus-untreated comparison; much of the interpretation came from comparisons with healthy controls.
Sam: That makes the overall metabolic story more secure than the explanation for the regional mismatch. Did they find any other spatial change?
Alex: Yes. Normally, fat droplets were concentrated near the central veins of small liver units. After treatment, more appeared toward the portal regions, where blood enters those units. The authors propose that local differences in fat uptake and use could explain that redistribution, but they did not establish the mechanism.
Sam: Could uneven drug delivery explain either the redistributed droplets or the differences between lobes?
Alex: They tested that using mass spectrometry imaging, which maps chemicals within tissue sections. The drug appeared evenly distributed, with similar signals in both lobes. That argues against uneven delivery, although the imaging experiments used only three animals per group.
Sam: Blood cholesterol improved too. Did cholesterol simply disappear from the liver as well?
Alex: No. Blood cholesterol fell toward healthy-control levels, while total liver cholesterol increased. The authors interpret the combination as likely increased liver uptake from blood, supported by transport-related gene changes. The balance between stored and free cholesterol also suggested improved handling, but uptake was not directly measured.
Sam: And how confidently can we attribute all these effects specifically to AMPK activation?
Alex: That remains unresolved here. ATX-304 also promotes mitochondrial uncoupling, where respiration becomes less tightly linked to making ATP, the cell’s energy currency. The authors propose that this could help explain energy expenditure and fewer oxidized, potentially harmful lipids, but this experiment does not separate those mechanisms.
Sam: Given the benefits and the unresolved regional harm, who should read the full study, and where should they start?
Alex: Researchers developing metabolic liver treatments, studying AMPK, or investigating spatial liver biology should read it. Start with the results on heterogeneous reductions in liver fat and fibrosis, especially the left-lobe versus right-median-lobe comparison. Then read the molecular analyses and discussion to separate measured changes from proposed explanations.
Sam: And what should everyone else carry away, given that this was a male-only mouse model rather than a human liver-disease trial?
Alex: This is evidence for further investigation, not established clinical benefit. Remember the central warning: a liver can improve on average while one region gets worse.
Sam: Thanks for listening.