Metabolomics of 3D cell co-culture reveals alterations in energy metabolism at the cross-talk of colorectal cancer-adipocytes

Andrea Corazzi Pelosi; Alex Ap Rosini Silva; Anna Maria Alves Piloto Fernandes; Pedro Paulo Menezes Scariot; Manoela Stahl Parisotto Oliveira; Andreia M Porcari; Denise Gonçalves Priolli; Leonardo Henrique Dalcheco Messias

doi:10.3389/fmed.2024.1436866

Metabolomics of 3D cell co-culture reveals alterations in energy metabolism at the cross-talk of colorectal cancer-adipocytes

Front Med (Lausanne). 2024 Oct 3:11:1436866. doi: 10.3389/fmed.2024.1436866. eCollection 2024.

Authors

Andrea Corazzi Pelosi¹, Alex Ap Rosini Silva², Anna Maria Alves Piloto Fernandes², Pedro Paulo Menezes Scariot¹, Manoela Stahl Parisotto Oliveira¹, Andreia M Porcari², Denise Gonçalves Priolli³, Leonardo Henrique Dalcheco Messias¹

Affiliations

¹ Research Group on Technology Applied to Exercise Physiology-GTAFE, Health Sciences Postgraduate Program, São Francisco University, Bragança Paulista, SP, Brazil.
² MS4Life Laboratory of Mass Spectrometry, Health Sciences Postgraduate Program, São Francisco University, Bragança Paulista, SP, Brazil.
³ Coloproctology Service of the Federal University of São Paulo, São Paulo and Faculty of Health Sciences Pitágoras de Codó, Codó, Brazil.

Abstract

Introduction: Colorectal cancer (CRC) is the third most incident and the second most lethal malignant tumor. Despite the recognized association between obesity and CRC, further clarification is necessary regarding the lipids that are overexpressed during the development of CRC. In this scenario, the combination of metabolomics and a three-dimensional (3D) co-culture model involving CRC tumor cells and lipids can enhance the knowledge of energy metabolism modifications at the cross-talk between colorectal cancer and adipocytes. This study aimed to screen potential metabolites in the three dimensional (3D) co-culture of CRC and adipocytes by investigating the metabolome composition of this co-culture released into the extracellular space, which is known as the secretome.

Methods: Pre-adipocyte cells (3T3-L1), human colon carcinoma (HT-29), and the 3D co-culture (3T3-L1 + HT-29) were cultured for the secretome obtention. Then, ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry (LC-HRMS) was employed to analyze the metabolomics of each secretome.

Results: Overall, 3.731 molecules were detected independent of the cell culture. When comparing the three cultures, 105 molecules presented a statistically significant difference in abundance between groups. Among these molecules, 16 were identified, with a particular emphasis on six lipids (PG 20:0, octadecenal, 3-Hydroxytetracosanoyl-CoA, 9,10-dihydroxy-octadecenoic acid, palmitoleic acid, and PA 18:4) and one amino acid derivative (acetylglutamic acid), which presented significant scores during the partial least-squares discriminant analysis (PLS-DA).

Discussion: Although it is too early to determine the possible impact of such molecules in a CRC microenvironment, these results open new avenues for further studies on the energy metabolism at the cross-talk of colorectal cancer adipocytes.

Keywords: bioinformatics; co-culture; metabolomics; neoplasm; rectum.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study was financed in part by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 (CAPES - 88887.504805/2020-000 AARS), São Paulo Research Foundation - FAPESP (Grants no. 2019/04314-6 APo and 2023/11638-8 AS), and the National Council for Scientific and Technological Development - CNPq (Process no. 408680/2021-0 LHDM) for the financial support.