Abstract 201: Establishment of 3D co-culture spheroid model for in- vitro drug screening

Abstract

Abstract Melanoma is an aggressive form of skin cancer that accounts for 75% of deaths of all skin cancers in the United States. Moreover, the number of cases is increasing at an alarming rate than any other cancer worldwide. But what is more alarming is the lack of chemo preventive or therapeutic drugs that are easily accessible to people and the absence of an efficient, effective and reliable model or platform for drug screening. A growing scientific and observational analyses suggests that antioxidants contain in blueberry inhibit tumor growth, and help slow down esophageal, lung, mouth, pharynx, endometrial, pancreatic, prostate, and colon cancers. Lately, the use of 3D multicellular spheroid culture has become a potential link to bridge the gap between monolayer culture and animal model. The treatment of blueberry extract (1mg) on the spheroids of A375 cells showed a disintegration of these spheroids and reduction in their growth. To optimize the culture conditions further herein, we used a novel three-dimensional (3D) coculture system with melanoma (A375), human fibroblast (HF-1) and Human umbilical endothelial cells (HUVECs) to study the effectiveness of drug in vitro. The multicellular nature of the coculture spheroid stimulates the microenvironment of the original tumor which makes it more efficient in drug screening process. The immunofluorescence assay on the 3D spheroid co-culture shows the interactions among these three cells which mimics the tumor microenvironment. We further analyzed the combination of blueberry extract and resveratrol which proved the effectiveness of blueberry extract as a potential chemosensitizer with resveratrol for skin cancer therapy. The complementary effect on the inhibition of A375 cell proliferation were observed when treated with 8 mg blueberry and 10 mM of resveratrol. The results of MTT assay indicates that the blueberry extracts showed potential cytotoxic activity against A375 cell line. 3D spheroid assay and 3D spheroid co-culture assays further confirmed the effectiveness of blueberry and resveratrol combination. Therefore, this platform could be applied for in vitro based drug screening studies. To conclude cytotoxic effects of the blueberry extract in human melanoma cells make them good candidates for further pharmacological studies to discover effective drugs for melanoma therapy. Citation Format: Christella J. Nelson, Vino T. Cheriyan, Abhishek Pandit, Joseph Francis, Alan Tackett. Establishment of 3D co-culture spheroid model for in- vitro drug screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 201.