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©The Author(s) 2025. Published by Baishideng Publishing Group Inc. All rights reserved.
Growth differentiation factor 11 modulates metabolism, mitigating the pro-tumoral behavior provided by M2-like macrophages in hepatocellular carcinoma-derived cells
Alejandro Escobedo-Calvario, Lisette Chávez-Rodríguez, Verónica Souza-Arroyo, Leticia Bucio-Ortiz, Roxana U Miranda-Labra, Felipe Masso, Araceli Páez-Arenas, Rogelio Hernández-Pando, Jens Marquardt, María Concepción Gutiérrez-Ruiz, Luis E Gomez-Quiroz
Alejandro Escobedo-Calvario, Lisette Chávez-Rodríguez, Verónica Souza-Arroyo, Leticia Bucio-Ortiz, Roxana U Miranda-Labra, María Concepción Gutiérrez-Ruiz, Luis E Gomez-Quiroz, Department of Ciencias de la Salud, Universidad Autónoma Metropolitana, Mexico City 09340, Ciudad de México, Mexico
Felipe Masso, Araceli Páez-Arenas, Unidad de Medicina Traslacional, Instituto Nacional de Cardiología Ignacio Chávez, Mexico City 14080, Ciudad de México, Mexico
Rogelio Hernández-Pando, Department of Pathology, National Institute of Medical Sciences and Nutrition Salvador Zubiran, Mexico City 14080, Ciudad de México, Mexico
Jens Marquardt, Department of Medicine, University of Lübeck, Lübeck 23552, Schleswig-Holstein, Germany
Co-corresponding authors: María Concepción Gutiérrez-Ruiz and Luis E Gomez-Quiroz.
Author contributions: Escobedo-Calvario A performed experiments; Escobedo-Calvario A, Chávez-Rodríguez L, Souza-Arroyo V, Bucio-Ortiz L, Hernández- Pando R, Marquardt J, and Gutiérrez-Ruiz MC contributed to methodology; Escobedo-Calvario A and Chávez-Rodríguez L contributed to investigation; Escobedo-Calvario A, Souza-Arroyo V, Bucio-Ortiz L, Masso F, Páez-Arenas A, and Gutiérrez-Ruiz MC contributed to writing; Souza-Arroyo V designed the biostatistical analysis; Bucio-Ortiz L obtained the funding; Miranda-Labra RU performed the reactive oxygen species studies; Masso F and Páez-Arenas A contributed to flow cytometry analysis; Hernández-Pando R and Marquardt J reviewed the manuscript; Gomez-Quiroz LE performed the conceptualization, project administration, initial draft preparation, and paper submission. All authors approved the final version of the article.
Institutional review board statement: The study was reviewed and approved by the Graduate Program in Experimental Biology at the Metropolitan Autonomous University.
Conflict-of-interest statement: All the authors report no relevant conflicts of interest for this article.
Data sharing statement: The data supporting the findings of this study are available from the corresponding author upon reasonable request.
Open Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Luis E Gomez-Quiroz, PhD, Department of Ciencias de la Salud, Universidad Autónoma Metropolitana Iztapalapa, No. 186 San Rafael Atlixco, Mexico City 09340, Ciudad de México, Mexico.
legq@xanum.uam.mx
Received: June 27, 2025
Revised: July 24, 2025
Accepted: September 23, 2025
Published online: October 28, 2025
Processing time: 122 Days and 12.3 Hours
BACKGROUND
Hepatocellular carcinoma (HCC) is one of the most aggressive tumors worldwide. Chronic inflammation contributes to tumor evolution, and the infiltration of tumor-associated macrophages (TAMs), also known as M2-like macrophages, is associated with the most aggressive behavior. Therefore, these macrophages provide the primary growth and migratory factors to the tumor cells, including those of HCC. Current therapies are not well optimized for eliminating transformed cells or neutralizing the tumor immune microenvironment leukocytes, such as TAMs. Growth differentiation factor 11 (GDF11) may represent a promising dual therapeutic target due to its reported anti-tumorigenic and immunomodulatory properties.
AIM
To characterize the effects of GDF11 in M2-like macrophages and the HCC cell interaction using a functional in vitro model.
METHODS
This research used THP-1 and Huh7 cell lines. We applied recombinant GDF11 (50 ng/mL) every 24 hours on THP-1 differentiated macrophages with M2-like polarization using interleukin-4 and interleukin-13. Firstly, the GDF11 effects on signaling, viability, proliferation, metabolism, and redox state in macrophages were characterized. Subsequently, we extracted conditioned media (CM) from macrophages and performed indirect co-cultures with Huh7 cells. The functional parameters were proliferation and migration assays. Finally, we characterized secretion in the CM using the cytokine array membrane assay.
RESULTS
The present study demonstrated that GDF11 activates the canonical pathway Smad2/3 without cytotoxic or proliferative effects. We provide evidence that GDF11 also diminishes the pro-tumoral properties of M2-like macrophages. GDF11 promoted the reduction of the M2-like macrophage marker, cluster of differentiation 206, indicating a loss of pro-tumoral properties in these leukocytes. Furthermore, this molecule induced changes in metabolism and an increase in reactive oxygen species. Using CM derived from GDF11-treated M2-like macrophages, we observed a reduction in the proliferation and migratory capacity of liver cancer cells. Moreover, the cytokine profile was affected by GDF11 stimulus, demonstrating that this molecule alters the pro-tumoral properties of TAMs, which in turn impact the behavior of HCC-derived cells.
CONCLUSION
This in vitro study suggests that mitigating tumor-promoting or M2-like macrophages with GDF11 may be an effective strategy for controlling the aggressiveness of HCC.
Core Tip: Hepatocellular carcinoma (HCC) is an aggressive tumor, and the infiltration of tumor-associated macrophages or M2-like macrophages increases aggressiveness. We need therapies to mitigate this pathology in the immune context. In the present study, our in vitro model demonstrates that growth differentiation factor 11 (GDF11) counteracts the pro-tumorigenic properties of M2-like macrophages. This was shown in the reduction of cluster of differentiation 206; furthermore, it induces changes in metabolism and the redox state. Conditioned media from GDF11-treated M2-like macrophages possessed changes in their cytokine profile that neutralized the proliferation and migration of HCC cells, demonstrating that GDF11 alters the pro-tumoral properties of macrophages. This study suggests that mitigating tumor-associated macrophages may be an effective strategy for controlling HCC.
