Abstract 7059: Remodeling of anaplastic thyroid cancer metabolic signature by natural alkaloid berberine
Authors: Jarboe Tara , DeSouza Nicole R. , Kopec Kaci , Geliebter Jan , Li Xiu-Min , Tiwari Raj K. 
Published: March 2024

Abstract
Anaplastic thyroid cancer (ATC) is a rare, fatal cancer with a five-year survival of 4%. Universally diagnosed at stage IV, ATC is characterized by its lack of differentiation and metabolic dysregulation. Refractory to all established therapies, we propose natural alkaloid berberine (BBR) as a therapeutic with multitarget efficacy to alter mitochondrial metabolism and reprogram ATC’s aggressive phenotype. Assays used monocyte cell line U937, ATC cell lines T238 and SW1736, and immortalized normal thyroid cell line Nthy-ori-3-1. All assays used 100µM concentration of BBR or DMSO vehicle control for 24 hours for a tumoricidal effect. Validation of in vitro findings via RNA-Seq was conducted by Genewiz from Azenta and Qiagen’s Ingenuity Pathway Analysis was used for in silico modeling. Mitochondria were isolated via differential centrifugation. Modeling the ATC tumor microenvironment, U937 cells were activated and polarized into a proinflammatory macrophage phenotype. BBR treatment at the activation/polarization stages, 33 soluble inflammatory mediators were downregulated in the conditioned media compared to controls. In targeting the aggressiveness of anaplastic thyroid disease, BBR slowed proliferation by 80% selectively in ATC cells from 48 to 72 hours, while sparing normal cells. BBR reduced migratory capacity by 30% in ATC cells after 24, 48, and 72 hours and reduced migration and invasion by 30%. These observations were substantiated by Western blot analysis - BBR selectively decreased phosphorylation of MEK, ERK, and ribosomal protein S6, crucial downstream regulators of the pro-proliferative and pro-survival pathways in ATC cells. Further, BBR specifically modulated cancer-associated metabolism as observed through an increase in AMPKα phosphorylation, a major rate-limiting protein in cancer-induced dysregulation with an anti-tumor effect. Validation of in vitro findings via RNA-Seq revealed more than 400 significantly differentially expressed genes involved in mitochondrial metabolism, glycometabolism, sirtuin signaling, apoptosis, and proliferation. Following a comprehensive analysis, we identified significant downregulation of 25 of 37 total mitochondrially encoded genes (MTs) and 13 of 13 mitochondrially encoded protein-coding genes comprising the oxidative phosphorylation complexes illuminating a clear link between BBR treatment and altered mitochondrial metabolism in ATC. Additionally, protein expression of significantly downregulated mitochondrial genes identified via RNA Seq was validated via Western blot demonstrating decreased mitochondrially-encoded protein expression related to oxidative phosphorylation. This work reveals a novel role for BBR as an inhibitor of mitochondrial metabolism that can be used to reprogram the aggressive nature of ATC and open the door for promising combination therapy in the treatment of ATC.
Citation Format: Tara Jarboe, Nicole R. DeSouza, Kaci Kopec, Jan Geliebter, Xiu-Min Li, Raj K. Tiwari. Remodeling of anaplastic thyroid cancer metabolic signature by natural alkaloid berberine [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 7059.