Interrogation of Breast Tumor-Vessel Interactions in a Tissue-Engineered Patient-Derived Biomimetic Platform
Xue Dong, Sarah Caughey, Jason Harris, Ryan Bender, Nabih Berri, Jason A. Spector
Laboratory of Bioregenerative Medicine & Surgery, Department of Surgery, Division of Plastic Surgery, Weill Cornell Medical College, New York, NY
Breast cancer (BC) is the most common cancer worldwide in women among all malignancies. As such, there is an urgent need for high fidelity in vitro platforms which closely resemble the local tumor microenvironment for the development of clinical therapeutics. Herein we created a tissue-engineered 3D biomimetic platform, derived from patient specific breast tissues that contains all components of the breast tumor microenvironment (glandular epithelial organoids, adipocytes, stromal vascular fraction (SVF)) within which are embedded engineered vascular channels. Hemispheroid triple negative breast tumor “buttons” were placed at predetermined distances between the vessels allowing for detailed studies of the tumor/vessel interactions. Polydimethylsiloxane (PDMS) molds were created using custom designed 3D-printed Poly-lactic-acid (PLA) molds (a Makerbot Replicator 5th Generation). The biomimetic platform was fabricated using adipocytes and other patient-derived tissue components mixed within neutralized 1% (w/v) Type I collagen to form the main structure in PDMS molds. RFP-MDA-MB-231 cells mixed with 1% collagen at a density of 40,000 cells/1.5uL were plated into the bulk to create the tumor hemispheroid “buttons”. The putative vascular channels within the same Z coordinate were seeded with RFP-smooth muscle cells (SMC) and GFP-endothelial cells (EC) sequentially after 24 hours of plating. Control constructs were made by generating the vascular structures within a collagen-only matrix. Constructs were cultured for 7, 14 and 21 days, confocal imaging and histological analysis followed. Patent vascular channels lined with SMC and EC were visualized 24 hours after seeding. H&E staining revealed successful fabrication of biomimetic platforms containing patient-derived adipocytes, SVF and ductal organoids. Fluorescent imaging and H&E staining showed SMC sprouts had formed and massively increased from vessels into surrounding ECM between 7 and 21 days of culture. Concurrently, there was a consistent EC neo-intimal monolayer formed within the channel. The hemispheroid tumor “buttons” were noted to increase in size and individual cells were seen invading preferentially towards the vessels to a much greater degree over time. Compared to collagen-only control group, the biomimetic group displayed increased tumor cell invasion on Day 7, 14 and 21. We have successfully engineered an advanced, patient-specific, vascularized biomimetic platform of the breast cancer microenvironment that not only replicates patient tissue characteristics, but also includes vascular structures and cancer hemispheres that closely resemble early tumors. Such a platform represents a highly potent tool that holds significant promise for diagnostic and therapeutic applications in the study of breast cancer.
Back to 2021 Abstracts