Impact of Multi-Mineral Intervention on the Proteome of Human Colon Organoids Under Pro- And Anti-Inflammatory Conditions

Abstract INTRODUCTION Aquamin, a multi-mineral supplement derived from fossilized red marine algae, has demonstrated benefits for colon barrier structure and function. Mesalamine (5-aminosalicylic acid; 5-ASA) is widely prescribed as maintenance therapy for patients with mild-to-moderate ulcerative colitis (UC) or those in remission. Our long-term objective is to assess whether Aquamin could be included in UC maintenance regimens by evaluating possible complementary effects or synergy with mesalamine and exploring any potential drug interactions. METHODS Human colon organoids were derived from healthy colon tissue and cultured under controlled conditions or subjected to a pro-inflammatory environment (by exposure to LPS and cytokines) to model mild-to-moderate UC. Organoids were then treated for 14 days with Aquamin alone, mesalamine alone, or a combination of both agents. Following the treatment period, proteomic analyses were performed to identify changes in protein expression induced by each treatment, both individually and in combination. RESULTS Distinct proteomic profiles were observed in colon organoids treated with Aquamin or mesalamine. Aquamin specifically enhanced the expression of proteins associated with the colon barrier, such as cadherin-17 and desmoglein-2. Mesalamine alone had minimal effect on these barrier components, and its co-administration with Aquamin did not alter Aquamin’s impact. In contrast, mesalamine alone upregulated several basement membrane proteins, with the combination of Aquamin and mesalamine eliciting a greater response than either agent individually. Moreover, mesalamine treatment led to upregulation of a range of inflammation-related proteins, including those involved in the complement and clotting/fibrinolytic cascades; these pro-inflammatory proteins were downregulated by Aquamin. Notably, when both agents were applied together, the proteomic changes in inflammation-associated proteins were more similar to the profile seen with mesalamine alone rather than Aquamin. The introduction of a pro-inflammatory stimulus further differentiated the responses to each intervention, with mesalamine more closely mirroring the effects of the pro-inflammatory challenge on inflammation-associated protein expression. CONCLUSION These findings indicate that Aquamin-induced upregulation of barrier proteins is not diminished by the simultaneous presence of mesalamine. There was also no evidence to suggest that Aquamin interferes with the capacity of mesalamine to modulate proteins involved in the anti-inflammatory response. Collectively, these results support the potential of Aquamin as a complementary agent in UC maintenance strategies, warranting further investigation into its combined use with established therapies.