X14 - Using 3D Spheroid Endometrial Cancer Models to understand the role of Adipose Stem Cells in Obesity Endometrial Cancer Tumorigenesis

Friday, October 10, 2025

10:00 AM - 11:00 AM PDT

Location: Exhibit Hall F, G & H

Presenting Author(s)

JV

Jasmine L. Vargas

Senior Undergraduate
UT San Antonio The University of San Antonio
Converse, Texas, United States

Co-Author(s)

FZ

Federico Zertuche

Graduate Student
University of Texas At San Antonio, United States

Primary Investigator(s)

Maria A. Gonzalez Porras

Assistant Professor
University of Texas San Antonio
San Antonio, Texas, United States

Introduction: : There has been an increase in endometrial cancer (EC) cases which have been linked to increasing obesity rates. The mechanisms linking obesity and EC remain undetermined. New findings suggest that adipose stem cells (ASCs) infiltrate the endometrial tumor microenvironment and have an influence on tumorigenesis as they secrete adipokines such as plasminogen activator inhibitor-1 (PAI-1), which has been found to exert paracrine influences on endometrial epithelial cells (EEC). Changes in extracellular matrix components such as integrins and collagen type I (COL-1) with ASC infiltration may play a role in the tumor microenvironment that may contribute to tissue remodeling and cancer progression and could be therapeutic targets. A 3D cell culture model was created to further understand the relationship between ASCs and EECs.

Materials and

Methods: : ASCs, non-oncogenic EECs (EME 6/7), and oncogenic EECs (KLE) were used to create spheroid models using the hang drop method. Monoculture and co-culture (1:1 ratio) spheroids were formed using ASCs and EEC cell lines with 10,000 total cells. Size analysis was done for all samples at different times over 3 days and 7 days by measuring spheroid area using ImageJ. qPCR was done to analyze expression of integrins, PAI-1, and COL-1.

Results, Conclusions, and Discussions:: Based on current data, size analysis showed that after 72 hours of spheroid formation, the average spheroid Areas were 0.129 mm^2 for ASC monoculture, 0.078 mm^2 for EME monoculture, 0.688 mm^2 for KLE monoculture, 0.102 mm^2 for ASC-EME co-culture, and 0.172 mm^2 for ASC-KLE co-culture. KLE monoculture spheroids displayed the largest size compared to all other groups. However, the addition of ASCs in co-culture led to a significant reduction in spheroid size, despite having the same total cell number. This suggests that ASC presence interferes with cancer cell–cell interactions and extracellular matrix (ECM) organization, likely through remodeling proteins such as collagen. When spheroid size was monitored over extended time points (96 hours, 144 hours, and 169 hours), minimal changes in spheroid size were observed, indicating that the size reduction occurred early during spheroid assembly. Given the current results, qPCR analysis revealed increased expression of PAI-1, integrin alpha 7 (ITA-7), and collagen I, in the ASC-KLE co-cultures, suggesting that ASC infiltration may alter ECM composition and disrupt cancer cell communication during spheroid formation. Gene expression analysis revealed an upregulation of ITA-7, PAI-1, and COL-1 in co-culture spheroids, suggesting that ASCs modulate signaling pathways within the endometrial tumor microenvironment. These changes indicate that ASCs may contribute to tumorigenic processes through paracrine and mechanical interactions. Notably, co-culture with KLE oncogenic cells resulted in smaller, more compact spheroids, which may reflect early extracellular matrix (ECM) remodeling and increased mechanical tension, hallmarks of tumor progression. The upregulation of ITA-7, a mechanosensitive integrin, further supports the involvement of ECM dynamics in these interactions. While direct assessment of ECM structure was not performed, the observed changes in current gene expression and spheroid area data imply that ASCs actively influence the 3D tumor architecture. This spheroid model therefore offers a physiologically relevant platform to study ASC–tumor interactions and their potential role in endometrial cancer progression.

Acknowledgements and/or References (Optional): :