TY - JOUR

T1 - Exploring the Heterogeneity of Cancer-Associated Fibroblasts via Development of Patient-Derived Cell Culture of Breast Cancer

AU - Ilyina, Anna

AU - Leonteva, Anastasia

AU - Berezutskaya, Ekaterina

AU - Abdurakhmanova, Maria

AU - Ermakov, Mikhail

AU - Mishinov, Sergey

AU - Kuligina, Elena

AU - Vladimirov, Sergey

AU - Bogachek, Maria

AU - Richter, Vladimir

AU - Nushtaeva, Anna

N1 - This research was funded by the grant of the State Program of the «Sirius» Federal Territory, «Scientific and technological development of the «Sirius» Federal Territory» (agreement no. 27-03 dated 27 September 2024).

PY - 2025/8/12

Y1 - 2025/8/12

N2 - Cancer-associated fibroblasts (CAFs) constitute a heterogeneous population of cells within the tumor microenvironment and are associated with cancer development and drug resistance. The absence of a universal classification for CAFs hinders their research and therapeutic targeting. To define CAF phenotypes, we developed patient-derived cell cultures of breast cancer (BC) and validated and characterized four distinct CAF subtypes (S1–S4) by Costa’s classification. Three out of five primary cell cultures of BC demonstrated different functional features rather than fixed cellular states due to the plasticity of the CAF phenotype. CAF crosstalk with cancer cells supported their survival in the presence of anticancer drugs. Based on the analysis of the cytotoxic effect of doxorubicin, cisplatin and tamoxifen, it was demonstrated that CAF-S4 and CAF-S1 cells were sensitive to the action of all drugs investigated, despite the fact that they possessed different mechanisms of action. CAF-S2 cells exhibited the highest level of resistance to the antitumour agents. Homotypic and heterotypic spheroids with CAFs could be used to model the fibrotic area of BC in vitro. The patient-derived cell cultures of CAFs formed spheroids. Hypoxia-activated CAF-S4 have been shown to stimulate the metastatic potential of triple-negative BC cells in a heterotypic spheroid model. Consequently, this study could be a starting point for the development of novel therapeutic strategies that target CAFs and their interactions with cancer cells.

AB - Cancer-associated fibroblasts (CAFs) constitute a heterogeneous population of cells within the tumor microenvironment and are associated with cancer development and drug resistance. The absence of a universal classification for CAFs hinders their research and therapeutic targeting. To define CAF phenotypes, we developed patient-derived cell cultures of breast cancer (BC) and validated and characterized four distinct CAF subtypes (S1–S4) by Costa’s classification. Three out of five primary cell cultures of BC demonstrated different functional features rather than fixed cellular states due to the plasticity of the CAF phenotype. CAF crosstalk with cancer cells supported their survival in the presence of anticancer drugs. Based on the analysis of the cytotoxic effect of doxorubicin, cisplatin and tamoxifen, it was demonstrated that CAF-S4 and CAF-S1 cells were sensitive to the action of all drugs investigated, despite the fact that they possessed different mechanisms of action. CAF-S2 cells exhibited the highest level of resistance to the antitumour agents. Homotypic and heterotypic spheroids with CAFs could be used to model the fibrotic area of BC in vitro. The patient-derived cell cultures of CAFs formed spheroids. Hypoxia-activated CAF-S4 have been shown to stimulate the metastatic potential of triple-negative BC cells in a heterotypic spheroid model. Consequently, this study could be a starting point for the development of novel therapeutic strategies that target CAFs and their interactions with cancer cells.

KW - breast cancer

KW - cancer-associated fibroblast

KW - fibrosis

KW - heterotypic 3D

KW - hypoxia

KW - patient-derived cell cultures

KW - primary cell culture

KW - spheroid

KW - stromal cells

KW - tumor microenvironment

UR - https://www.scopus.com/pages/publications/105014242546

UR - https://www.mendeley.com/catalogue/90b27bb7-a745-3c52-92d0-648c4ae88f1e/

U2 - 10.3390/ijms26167789

DO - 10.3390/ijms26167789

M3 - Article

C2 - 40869109

VL - 26

JO - International Journal of Molecular Sciences

JF - International Journal of Molecular Sciences

SN - 1661-6596

IS - 16

M1 - 7789

ER -