Patient-Derived Tumor Chemosensitization of GKB202, an Antrodia Cinnamomea Mycelium-Derived Bioactive Compound

Molecules2021,26, 6018. https://doi.org/10.3390/molecules26196018

Abstract

Oral cancers, hepatocellular carcinoma, and colorectal cancers are the three most common cancers, leading to 18,000 cases of cancer-related mortality in Taiwan per year. To bridge the gap towards clinical translation, we developed a circulating tumor cell (CTC) organoid culture workflow that efficiently expands CTC from patients to test Antrodia Cinnamomea mycelium-derived bioactive compounds. Three ACM-derived bioactive compounds were evaluated for tumor chemosensitization characteristics. Significant and consistent cytotoxic/5-FU sensitizing effects of GKB202 were found on 8 different patient-derived tumors. Acute toxicity profile and hepatic metabolism of GKB202 in rats suggest GKB202 is rapidly cleared by liver and is well tolerated up to the dose of 20 mg/kg. This comprehensive study provides new evidence that liquid fermentation of Antrodia cinnamomea mycelium (ACM) contains bioactive compounds that lead to effective control of CTC, especially when combined with 5-FU. Together, these data suggest ACM-derived GKB202 may be considered for further clinical investigation in the context of 5-FU-based combination therapy.

Figure 1. (A) HPLC of purified GKA201, GKB202, and GKC203 from ACM with retention time labeled at 5.063 for GKA201, 5.420 for GKB202, 7.285 min for GKC 203. (B) LC-QTOF/MS spectra of GKA201 with parent ion detected at m/z 313.14481; GKB202 with parent ion detected at m/z 463.26310; GKC203 with parent ion detected at m/z 330.16998.

Figure 2. Immunofluorescence analysis of expanded circulating tumor cells. OC: oral cancer; HCC: hepatocellular carcinoma; CRC: colorectal cancer. PanCK is a cancer-specific surface marker. CD45, GPC3, EGFR are markers specific to leucocytes, HCC, and CRC, respectively. Representative images from one patient in each cancer type is shown. The scale bar is 75 μm in length.

Figure 3. Ex vivo expansion toxicity assay with different purified AC compounds at low and high dosage (4 and 20 µg/mL respectively) in different cancer types. Each patient-derived CTC was tested three times for different assay groups. (A) OC: oral cancer, total patients, N = 2; (B) HCC: hepatocellular carcinoma, total patients, N = 3; © CRC: colorectal cancer, total patients N = 3. * Represents a significant difference p < 0.05 and ** represents a significant difference p < 0.01.

Figure 4. GKB202 remaining versus time profile in SD rat Liver S9. Data is presented in mean values of triplicate measurements.