Developing an Approach for Evaluating the Cardiotoxic Potential of Botanicals

Julie Krzykwa¹Hemantkumar S Chaudhari²Andre Monteiro Da Rocha³Matthias Gossmann⁴Peter Hoffmann⁵Yaser Khokhar⁶Nathan Meyer⁷Jin Young K Park⁸Robert Sprando⁹Ravi Vaidyanathan¹⁰Remco HS Westerink¹¹Joseph C Wu⁶Jeffrey Yourick¹²Shane Rui Zhao⁶Constance A Mitchell^1*

• ¹Health and Environmental Sciences Institute (HESI), Washington, D.C., United States
• ²L’Oréal Research & Innovation, Clark, United States
• ³Frankel Cardiovascular Center Cell Regeneration Core/Internal Medicine-Cardiology, University of Michigan, Ann Arbor, United States
• ⁴innoVitro GmbH,, Juelich, Germany
• ⁵Consultant,, Beaufort, United States
• ⁶Stanford Cardiovascular Institute, Stanford University, Stanford, United States
• ⁷FUJIFILM Cellular Dynamics,, Madison, United States
• ⁸U.S. Food and Drug Administration, Human Foods Program, Office of Food Chemical Safety, Dietary Supplements and Innovation, College Park, United States
• ⁹U.S. Food and Drug Administration, Human Foods Program, Office of Chemistry and Toxicology,, Laurel, United States
• ¹⁰FUJIFILM Cellular Dynamics, Inc., Madison, United States
• ¹¹Neurotoxicology Research Group, Division of Toxicology, Institute for Risk Assessment Sciences (IRAS), Faculty of Veterinary Medicine, Utrecht University, Utrecht, Netherlands
• ¹²U.S. Food and Drug Administration, Human Foods Program, Office of Chemistry and Toxicology, Laurel, United States

Botanicals (e.g., extracts derived from plants, algae, or fungi) are increasingly utilized by consumers with 2 the hope of enhancing their health, managing symptoms, or preventing ailments; however, these 3 products have often had limited pre-market toxicity testing. Traditional toxicity testing (e.g., rodent 4 testing) is complicated by the nature of botanicals as complex mixtures and the potential for lot-to-lot 5 variability in chemical constituents. Cardiotoxicity is a key area of concern, as adverse effects on the 6 cardiovascular system can have severe consequences, and although not commonly reported, there have 7 been reports of adverse cardiac events. New approach methodologies (NAMs) offer human-relevant, 8 efficient, innovative, and cost-effective solutions for evaluating the cardiotoxicity of botanicals. The 9 Botanical Safety Consortium (BSC) was established to focus on identifying suitable NAMs to screen for 10 potential toxicities associated with these widely used products. This manuscript outlines the BSC 11 Cardiotoxicity Working Group's approach for evaluating NAMs for assessing the potential cardiotoxicity 12 of botanicals. These NAMs leverage in vitro models, such as human-induced pluripotent stem cell-13 derived cardiomyocytes, and techniques like microelectrode arrays, voltage and calcium optical 14 mapping, contractile force measurement, and mitochondrial function assays to evaluate botanical-15 induced effects on the cardiovascular system. Using well-characterized botanical extracts as case 16 studies, the BSC aims to refine a toolkit for high-throughput and human-relevant cardiotoxicity 17 screening. This foundational work supports the broader goal of improving botanical safety assessment 18 practices and advancing the application of NAMs in regulatory toxicology.