Abstract

Biomimetic Tissue-Engineered Technology to Identify Tumor-Stroma Cross Talk and Novel Drug Candidates

Tania Konry, Assistant Professor, Northeastern University

Development of a physiologically relevant and reproducible in vitro cancer model for preclinical and pharmacological studies continues remains a critical unmet need. Two-dimensional (2D) tissue culture models and monolayer cultures lack realistic complexity of TME and tumor heterogeneity, while the current methods of ex vivo 3D tumor models suffer from number of drawbacks including a) inconsistencies related to achieving uniformity of spheroid size, cell–cell and cell–matrix interactions, b) not suited for high-throughput screening assays and c) the difficulty to limit shear-related cell damage. A variety of microfluidic device designs have been utilized for long-term tumor spheroid culture and drug efficacy evaluation, however, most lack the integral TME and dynamic perfusion and/or influence immune-tumor cross-talk or/and prolong culture capabilities.  Our innovative microfluidic platform technology is focused to generate biomimetic multicellular immunogenic spheroids. The proposed technology is based on integrated high-throughput microfluidic droplet based device capable of sequential generation, maintenance via continuous perfusion, stimulation and monitoring cellular dynamics of Alginate (Alg)-doped based micronized tumor spheroids. The Alg - doped hydrogel provides the mechanical strength to maintain the structural integrity of the spheroids and permit cellular encapsulation to mimic several important aspects of the in vivo environment such as a synthetic extracellular matrix (ECM), which promotes attachment of stroma cellular integrins to initiate signaling for cell survival and simulate the TME. We examined its capability to (1) allow recreation of TME and pathophysiological gradients to mimic the resistance to drug penetration; (2) cell composition and heterogeneity of the  tumor to evaluate tumor cell survival in real-time; (3) investigate spatiotemporal interaction between cancer-stroma and cancer-immune cells during therapy; and (4) capture the dynamic data such as secretome analyses from lymphocyte and stroma cells that exert paradoxical effects in the TME, changes in cellular proliferation, and viability and migration in the presence and absence of immune and stromal cells. We hypothesize that a precisely defined spheroid size and composition at the onset of treatment will clearly affect and predict response to treatment. Our platform provides a novel yet overall simple in vitro platform that will generate and analyze 3D monodisperse hydrogel tumors for biologic studies and therapeutic drug screening, which is a critical step in the translational cancer drug development.