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Proteomic Profile of Human Colon Organoids: Effects of a multi-mineral intervention alone and in the presence of pro-Inflammatory and anti-inflammatory treatments

ABSTRACT: Introduction: Aquamin, a multi-mineral product derived from fossilized red marine algae, has been shown to improve colon barrier structure and function. Mesalamine is commonly used as maintenance therapy for patients with mild-to-moderate ulcerative colitis (UC) or those in remission. Our long-term aim is to evaluate if Aquamin can be part of a UC maintenance regimen, examining potential complementary efficacy or synergy with Mesalamine, as well as any possible drug interactions. Methods: Human colon organoids were maintained under controlled conditions or exposed to a pro-inflammatory stimulus to mimic the environment in mild-to-moderate UC. Organoids were treated with Aquamin alone, Mesalamine alone, or the two agents in combination for 14 days. At the end of the treatment period, proteomic analysis was conducted to evaluate protein changes induced by the two agents (individually and in combination). Results: Colon organoids treated with Aquamin or Mesalamine exhibited distinct protein expression profiles. Aquamin enhanced the expression of colon barrier proteins (e.g., cadherin-17 and desmoglein-2). Mesalamine by itself had minimal impact on these moieties; when present with Aquamin, it did not alter Aquamin’s response. By itself, treatment with Mesalamine alone resulted in up-regulation of basement membrane proteins; the combination of Aquamin and Mesalamine was more effective than either alone. In contrast to these results, Mesalamine up-regulated numerous proteins directly related to inflammation including members of the complement and clotting/fibrinolytic cascades. These were down-regulated with Aquamin. When the two agents were utilized in combination, changes in the expression of inflammation-related proteins resembled the profile seen with Mesalamine alone more than the profile obtained with Aquamin. Of interest, the presence of a pro-inflammatory stimulus further highlighted the unique responses to the two interventions, with Mesalamine aligning more closely with the pro-inflammatory stimulus in its effect on the expression of inflammation-associated proteins. Conclusion: The data presented here suggest that the induction of barrier proteins by Aquamin would not be counteracted by the concomitant presence of Mesalamine. The current studies also found no evidence to suggest that the presence of Aquamin would interfere with the capacity of Mesalamine to alter the expression of proteins that are part of the anti-inflammatory shield.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Colon Epithelial Cell, Colon

DISEASE(S): Ulcerative Colitis

SUBMITTER: Muhammad Aslam

LAB HEAD: Muhammad N Aslam

PROVIDER: PXD062086 | Pride | 2026-03-16

REPOSITORIES: Pride

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			Action	DRS
	PRF_A_2023_J_VARA_1130_64479_64486_1.raw	Raw
	PRF_A_2023_J_VARA_1130_64479_64486_2.raw	Raw
	PRF_A_2023_J_VARA_1130_64479_64486_3.raw	Raw
	PRF_A_2023_J_VARA_1130_64479_64486_4.raw	Raw
	PRF_A_2023_J_VARA_1130_64479_64486_5.raw	Raw

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Project description:Background. Recent studies demonstrated that Aquamin®, a calcium-, magnesium-, and multiple trace element-rich natural product, improves barrier structure and function in colonoids obtained from the tissue of healthy subjects. The goal of the present study was to determine if the colonic barrier could be improved in tissue from subjects with ulcerative colitis (UC). Methods. Colonoid cultures were established with colon biopsies from 9 individuals with UC. The colonoids were then incubated for a 2-week period under control conditions (i.e., in culture medium with a final calcium concentration of 0.25 mM) or in the same medium supplemented with Aquamin® to provide 1.5 – 4.5 mM calcium. Effects on differentiation and barrier improvement were determined using several approaches: phase-contrast & scanning electron microscopy, quantitative histology & immunohistology, mass spectrometry-based proteome assessment and transmission electron microscopy. Results. While there were no gross changes in colonoid appearance, there was evidence of increased differentiation (larger lumen diameter and wall thickness) and greater expression of cytokeratin 20 (CK20) and lesser expression of Ki67 with the intervention. Aquamin®-treated colonoids demonstrated a modest up-regulation of tight junctional proteins but stronger induction of adherens junction proteins and desmosomal proteins. Increased desmosomes were seen at the ultrastructural level. Proteomic analysis also demonstrated increased expression of basement membrane proteins and hemidesmosomal components. Proteins expressed at the apical surface (mucins and trefoils) were also increased as were several additional proteins with anti-microbial activity or that modulate inflammation. Similarly, growth and cell cycle regulatory proteins (e.g., Ki67, nucleophosmin, and stathmin) were significantly down-regulated. Laminin interactions, fibronectin matrix formation and extracellular matrix organization were the top three up-regulated pathways with the intervention. Conclusion. A majority of individuals including patients with UC do not reach the recommended daily intake for calcium and other minerals. To the extent that such deficiencies might contribute to the weakening of the colonic barrier, the findings employing UC tissue-derived colonoids here suggest that adequate mineral intake might improve the colonic barrier

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Proteomic Profile of Human Colon Organoids: Effects of a multi-mineral intervention alone and in the presence of pro-Inflammatory and anti-inflammatory treatments

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