SCFA Boosts HCC Therapy by Regulating Intracellular Calcium Homeostasis

In the evolving landscape of cancer treatment, the role of gut microbiota has garnered significant attention, particularly the impact of short-chain fatty acids (SCFAs) such as butyric, propionic, and acetic acid on hepatocellular carcinoma (HCC) therapy. This article delves into the groundbreaking research published in Hepatology in 2023, which highlights how gut microbial metabolite butyrate can enhance anticancer therapy by regulating intracellular calcium homeostasis. 

Key Role of Short-Chain Fatty Acids in Improving Hepatocellular Carcinoma Treatment

The gut microbiota is recognized as crucial in cancer therapy, as suppressing their metabolism with antibiotics can reduce the effectiveness of antitumor treatments. Conversely, supplementation with commensal bacteria can improve the response of cancer to therapy. However, the underlying mechanisms remain unclear. Short-chain fatty acids (SCFAs), including acetic acid, propionic acid, and butyric acid, are the most abundant gut microbial metabolites and have demonstrated immunomodulatory potential in various autoimmune, inflammatory, and cancer models. Prior gut microbiome studies have shown a decrease in butyrate-producing bacterial species in patients with human hepatocellular carcinoma (HCC). Whether SCFAs can enhance the anticancer therapeutic effect of HCC was elaborated in a research paper titled "Gut microbial metabolite butyrate improves anticancer therapy by regulating intracellular calcium homeostasis" published in Hepatology in 2023. MetwareBio provided targeted detection services for short-chain fatty acids in this study.

The butyrate metabolism is activated in the plasma of HCC patients

Using SCFA-targeted metabolomic profiling, butyrate levels were examined in both HCC patients and healthy individuals, revealing that butyrate levels in the plasma of HCC patients were lower than those in healthy individuals. Previous reports have indicated that, through 16S rRNA gene sequencing, certain butyrate-producing bacterial families were found to be depleted in early HCC patients. Combined with the GMrepo database (https://gmrepo.humangut.info), it was discovered that the abundances of primary butyrate-producing bacteria in the feces of patients with liver diseases, including Eubacterium rectale, Roseburia hominis, and Bifidobacterium, were significantly lower than those in healthy individuals. These findings suggest that butyric acid metabolism is activated in HCC patients, resulting in decreased butyrate levels.

The SCFA butyrate  inhibits cell proliferation, colony formation, and migrationin HCC

In vitro culture experiments of HCC cell lines HCCLM3 and MHCC97H revealed a dose-dependent inhibitory effect of three SCFAs on HCC cell growth. Cell migration analysis demonstrated that, compared to the control group, butyrate (NaBu) treatment significantly reduced the migratory capacity of HCCLM3 and MHCC97H cells, respectively. This phenomenon was not observed in cells treated with the same concentrations of sodium acetate or sodium propionate. In summary, the results indicate that butyrate (NaBu) exhibits anticancer effects in vitro.

Butyrate treatment upregulated calcium signaling pathways and promoted ROS production.

RNA-Seq analysis revealed a significant enrichment of "calcium signaling pathways" after NaBu treatment, suggesting that NaBu exerts anti-proliferative and anti-metastatic effects in HCC by regulating intracellular calcium signaling. Further experiments demonstrated that NaBu activates calcium signaling gene transcription by promoting H3K9 acetylation and binding to their promoters. These results indicate that NaBu exerts tumor-suppressive effects by increasing gene transcription in calcium signaling pathways and promoting ROS production.

The supplementation of butyrate improves the therapeutic effect of HCC

Based on the aforementioned results, researchers treated cells with increasing concentrations of both NaBu and sorafenib at a fixed ratio, leading to a significant increase in apoptotic cells. When compared to treatment with NaBu or sorafenib alone, the combination of NaBu and sorafenib strongly inhibited cell proliferation in MHCC97H cells. Additionally, the study developed an anti-HCC drug combination of sodium butyrate and sorafenib, designated as BS@PEAL-GPC3 NPs, and validated its therapeutic effect in a mouse model of liver cancer. BS@PEAL-GPC3 NPs significantly reduced HCC progression, demonstrating excellent targeting efficiency and safety for HCC.

This study demonstrates that the gut microbial metabolite butyric acid enhances anticancer therapeutic effects by regulating intracellular calcium homeostasis. Gas chromatography-mass spectrometry (GC-MS) analysis revealed lower butyrate levels in the plasma of HCC patients. Supplementation with butyrate or deletion of the ACADS gene, encoding a key enzyme in butyrate metabolism, significantly inhibited HCC proliferation and metastasis. Analysis of the gene expression profiles upregulated by butyrate supplementation or ACADS knockout revealed the activation of calcium signaling pathways, leading to disruption of intracellular calcium homeostasis and the generation of reactive oxygen species (ROS). The supplementation of butyrate potentiated the therapeutic effect of the tyrosine kinase inhibitor sorafenib. Based on these findings, researchers developed mPEG-PLGA-PLL nanoparticles coated with anti-gpc3 antibodies and encapsulating both butyrate and sorafenib. These nanoparticles prolonged drug retention time and enhanced drug targeting, thereby achieving a higher anticancer effect. In summary, this study provides novel insights into the mechanism of gut microbial metabolites inhibiting HCC progression and offers a transformative approach to improving the targeted therapeutic efficacy of clinical cancer treatment.

Conclusion

The pioneering study on the role of gut microbial metabolites, particularly butyric acid, in enhancing the efficacy of anticancer therapies represents a significant leap forward in the field of oncology. 

Short-chain fatty acids (SCFAs), also known as volatile fatty acids (VFAs), are metabolites produced by gut microbiota. SCFAs generated by the gut microbiota can directly regulate host health and exert their functions through energy regulation, intestinal mucosal barrier maintenance, immune modulation, and the induction of tumor cell differentiation and apoptosis. 

Metwarebio utilizes a GC-MS platform and a self-constructed database to perform absolute quantitative detection of 11 types of short-chain fatty acids simultaneously. This method employs a dual verification approach with internal and external standards, providing broad coverage and accurate quantification, suitable for various diseases related to short-chain fatty acids as well as studies on the interaction between gut microbiota and the host. For more detailed information, please visit https://www.metwarebio.com/short-chain-fatty-acids/.

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