ABC99

Cell lines

Human stem cells from the apical papilla (hSCAPs)

Preparation Method

Human stem cells from the apical papilla (hSCAPs) were recovered and cultured in a cell culture incubator under the conditions of 37°C with 5% CO₂ (v/v). The culture medium was refreshed every 3 days. The normal proliferation medium (PM) contained α-MEM, 10% fetal bovine serum, and 1% penicillin. When cells reached 80% confluency, odontoblastic medium (OM) contained α-MEM, 10% fetal bovine serum, 50mg/L L-Ascorbic acid, 5mmol/L sodium β-phosphoglycerate, and 10nmol/L dexamethasone, was used to induce odontoblastic differentiation. To down-regulate the extracellular NOTUM level, ABC99, a small molecule inhibitor of NOTUM, was added to the OM medium at 2ug/ml. Cells were collected on odontoblastic induction day 3, 7, and 14. Alkaline phosphatase (ALP) staining, Alizarin red (ARS) staining and qRT-PCR were performed according to the manufacturer’s Instructions.

Reaction Conditions

2µg/mL; 7–14 days

Applications

ABC99 suppressed odontoblastic differentiation of human stem cells from apical papilla (hSCAPs), reduced ALP activity, blocked mineralized nodule formation, and decreased DSPP, DMP1 and ALP mRNA levels.

Animal models

C57BL/6J mice

Preparation Method

8-week-old, specific-pathogen-free C57BL/6J mice were kept in an air-conditioned room at a temperature of 23±1℃, a relative humidity of 50–70%, and a 12h light/dark cycle. APKS tumor organoids were orthostatic implanted into the colonic mucosa of C57BL/6J mice under endoscopic guidance to establish an in situ colorectal cancer model. Mice were treated with ABC99(10mg/kg/day; p.o.) or vehicle (corn oil) control after 4 weeks of engraftment and tumour growth. Mice are treated for indicated number of weeks (week 0-4 weeks post-implantation) (n=7 mice). Tumour weight, number of macrometastases in the liver and number of lymph node metastases in mice were qualified. Bioluminescent imaging (BLI) and Kaplan–Meier survival analysis were performed to monitor tumor burden and compare survival. 

Dosage form

10mg/kg/day; p.o.; 4 weeks

Applications

ABC99 almost completely prevented liver metastasis and lymph-node metastasis, reduced primary tumor weight in orthotopic colorectal cancer mice.

References:
[1] Zhao Q, Ren H, Wang N, Yuan X, Zhao Y, Wen Q. NOTUM plays a bidirectionally modulatory role in the odontoblastic differentiation of human stem cells from the apical papilla through the WNT/?-catenin signaling pathway. Arch Oral Biol. 2024;160:105896.
[2] Tian Y, Wang X, Cramer Z, et al. APC and P53 mutations synergise to create a therapeutic vulnerability to NOTUM inhibition in advanced colorectal cancer. Gut. 2023;72(12):2294-2306.