Studying Tumor Angiogenesis and Cancer Invasion in a Three‐Dimensional Vascularized Breast Cancer Micro‐Environment

Abstract

Metastatic breast cancer is one of the deadliest forms of malignancy, primarily driven by its characteristic micro‐environment comprising cancer cells interacting with stromal components. These interactions induce genetic and metabolic alterations creating a conducive environment for tumor growth. In this study, a physiologically relevant 3D vascularized breast cancer micro‐environment is developed comprising of metastatic MDA‐MB‐231 cells and human umbilical vein endothelial cells loaded in human dermal fibroblasts laden fibrin, representing the tumor stroma. The matrix, as well as stromal cell density, impacts the transcriptional profile of genes involved in tumor angiogenesis and cancer invasion, which are hallmarks of cancer. Cancer‐specific canonical pathways and activated upstream regulators are also identified by the differential gene expression signatures of these composite cultures. Additionally, a tumor‐associated vascular bed of capillaries is established exhibiting dilated vessel diameters, representative of in vivo tumor physiology. Further, employing aspiration‐assisted bioprinting, cancer–endothelial crosstalk, in the form of collective angiogenesis of tumor spheroids bioprinted at close proximity, is identified. Overall, this bottom–up approach of tumor micro‐environment fabrication provides an insight into the potential of in vitro tumor models and enables the identification of novel therapeutic targets as a preclinical drug screening platform.

3D Bioprinted perfusable and vascularized breast tumor model for dynamic screening of chemotherapeutics and CAR-T cells

“…Collagen and fibrin, both being abundantly present at a native tumor site, were incorporated in our study as the bioprinting substrate. Building on our previous study (20) , we incorporated 2 mg/ml collagen along with 3 mg/ml fibrinogen in this study (C2F3), to build the tumor microenvironment. In order to precisely position H231F spheroids in the composite C2F3 hydrogel, it was essential to realize the entire bioprinting process prior to complete crosslinking of C2F3.…”

3D bioprinting technology for modeling vascular diseases and its application

“…3D bioprinting provides an approach for the controlled printing or seeding of tumor cells or spheroids with ECs and fibroblasts, understanding the study of interactions between angiogenesis and the TME. Additionally, 3D bioprinting enables the fabrication of multilayer hydrogel structures that mimic human physiological scale, supporting the study of angiogenesis complexity and cancer-EC interactions [195][196][197][198][199][200].…”

Tumoural Hypoxic Extracellular Vesicles Foster a Protective Microenvironment in Triple‐Negative Breast Cancer

“…A significant limitation of multicellular 3D co‐cultures is the difficulty of evaluating responses in individual cells from the cultured mass, which often relies upon more refined and less accessible techniques (Dey et al. 2021 ). To circumvent this issue, we described a novel method to study the TME in vitro where cells are cultured in tandem in a commercially available system, henceforth named a multicellular circulating co‐culture (MC‐CC).…”

“…The recent development of techniques such as multicellular co-cultures, microfluidics and 3D culture has improved the complexity with which we can investigate basic mechanisms (Casali et al 2024;Park et al 2022;Rebelo et al 2018). A significant limitation of multicellular 3D cocultures is the difficulty of evaluating responses in individual cells from the cultured mass, which often relies upon more refined and less accessible techniques (Dey et al 2021). To circumvent this issue, we described a novel method to study the TME in vitro where cells are cultured in tandem in a commercially available system, henceforth named a multicellular circulating co-culture (MC-CC).…”