TY - JOUR

T1 - The Activity of Human NK Cells Towards 3D Heterotypic Cellular Tumor Model of Breast Cancer

AU - Leonteva, Anastasia

AU - Abdurakhmanova, Maria

AU - Bogachek, Maria

AU - Belovezhets, Tatyana

AU - Yurina, Anna

AU - Troitskaya, Olga

AU - Kulemzin, Sergey

AU - Richter, Vladimir

AU - Kuligina, Elena

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 - 2025/7/8

Y1 - 2025/7/8

N2 - Due to the complexity of modeling tumor-host interactions within the tumor microenvironment in vitro, we developed a 3D heterotypic cellular breast cancer (BC) model. We generated spheroid models using MCF7, MDA-MB-231, and SK-BR-3 cell lines alongside cancer-associated (BrC4f) and normal (BN120f) fibroblasts in ultra-low attachment plates. Stromal spheroids (3Df) were formed using a liquid overlay technique (graphical abstract). The YT cell line and peripheral blood NK (PB-NK) cells were used as immune components in our 3D model. In this study, we showed that stromal cells promoted tumor cell aggregation into spheroids, regardless of the initial proliferation rates, with NK cells accumulating in fibroblast-rich regions. The presence of CAFs within the model induced alterations in the expression levels of MICA/B and PD-L1 by tumor cells within the 3D-2 model. The feasibility of utilizing a 3D cell BC model in combination with cytokines and PB-NKs was evaluated. We observed that IL-15 and IL-2 enhanced NK cell activity within spheroids, whereas TGFβ had varying effects on proliferation depending on the cell type. Stimulation with IL-2 and IL-15 or TGFβ1 altered PB-NK markers and stimulated their differentiation into ILC1-like cells in 3D models. These findings underscore the regulatory function of CAFs in shaping the response of the tumor microenvironment to immunotherapeutic interventions.

AB - Due to the complexity of modeling tumor-host interactions within the tumor microenvironment in vitro, we developed a 3D heterotypic cellular breast cancer (BC) model. We generated spheroid models using MCF7, MDA-MB-231, and SK-BR-3 cell lines alongside cancer-associated (BrC4f) and normal (BN120f) fibroblasts in ultra-low attachment plates. Stromal spheroids (3Df) were formed using a liquid overlay technique (graphical abstract). The YT cell line and peripheral blood NK (PB-NK) cells were used as immune components in our 3D model. In this study, we showed that stromal cells promoted tumor cell aggregation into spheroids, regardless of the initial proliferation rates, with NK cells accumulating in fibroblast-rich regions. The presence of CAFs within the model induced alterations in the expression levels of MICA/B and PD-L1 by tumor cells within the 3D-2 model. The feasibility of utilizing a 3D cell BC model in combination with cytokines and PB-NKs was evaluated. We observed that IL-15 and IL-2 enhanced NK cell activity within spheroids, whereas TGFβ had varying effects on proliferation depending on the cell type. Stimulation with IL-2 and IL-15 or TGFβ1 altered PB-NK markers and stimulated their differentiation into ILC1-like cells in 3D models. These findings underscore the regulatory function of CAFs in shaping the response of the tumor microenvironment to immunotherapeutic interventions.

KW - 3D cancer cell models

KW - breast cancer

KW - cancer-associated fibroblast

KW - co-culture

KW - immunotherapy

KW - natural killer cell

KW - patient-derived cell culture

KW - spheroids

KW - stromal cell

KW - tumor microenvironment

UR - https://www.mendeley.com/catalogue/0a9a4005-9769-3eea-bd8f-cba4dc288a66/

UR - https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=105011651090&origin=inward

U2 - 10.3390/cells14141039

DO - 10.3390/cells14141039

M3 - Article

C2 - 40710292

VL - 14

JO - Cells

JF - Cells

SN - 2073-4409

IS - 14

M1 - 1039

ER -