Engineered and Instrumented Three-Dimensional Tumor-Immune Model System

Elizabeth Wheeler | 19-SI-003

Project Overview

Cancer is a complex, multi-scale, systemic disease that has continued to elude cure. One of the greatest challenges in developing predictive cancer models and therapeutic screens is the lack of adequate in vitro models that accurately reflect in vivo tumor behavior and can provide quantitative multi-scale data to define the complex relationship between tumor composition, microenvironment, cellular signaling and functional outcomes. The overall goal of this project was to develop an instrumented ex vivo 3D tumor model that enables spatio-temporal control and measurement of multiple tissue, cellular and molecular components that drive tumor growth. Our approach was to bioprint a tumor/microenvironment-immune construct instrumented with sensors (ranging from genetic to macro scale) to study tumor physiology, drug response and evolution. In addition, we measured biophysical attributes of the structural environment of the tumor such as stiffness, pH, and oxygen tension to map out tumor architecture. This research will advance understanding of the role of tumor-immune crosstalk on drug response and resistance by identifying the cellular pathways that determine whether or not tumors grow and are recognized by the immune system.

Using our engineered multiwell bioreactor platform we were able to increase the throughput of experiments and generate gradients in chemotherapy concentration better mimicking transport in tumors. While performing chemotherapeutic dosing of human derived tumoroids in the platform for comparison to other in vitro tests, we demonstrated the use of a non-destructive metabolic assay for monitoring organoid health over time as well as monitored the interplay between tumor and extracellular matrix. In addition, we demonstrated that the extracellular matrix can alter tumor-immune interactions. Throughout the lifetime of this project, we correlated cellular, genomic and proteomic datasets for future computational prediction and discovery studies

Mission Impact

This project supports the NNSA strategic goal of the advancement of the science, technology, and engineering competencies that are the foundation of the NNSA mission, as well as the expansion and application of our science and technology capabilities to deal with broader national security issues. This project also supports Lawrence Livermore National Laboratory's core competency in bioscience and bioengineering and addresses its chemical and biological countermeasures research and development challenge.

Publications, Presentations, and Patents

Sebastian, Aimy, Hum, Nicholas, Martin, Kelly, Gilmore, Sean, Peran, Ivana, Byers, Stephen, Wheeler, Elizabeth, Coleman, Matthew and Loots, Gabriela. 2020. "Single-Cell Transcriptomic Analysis of Tumor-Derived Fibroblasts and Normal Tissue-Resident Fibroblasts Reveals Fibroblast Heterogeneity in Breast Cancer." Cancers. 12: 1307.

Martin, Kelly, Hum, Nicholas, Sebastian, Aimy, He, Wei, Siddiqui, Salma, Ghosh, Paramita, Pan, Chong-xian, White, Ralph and Gabriela G. Loots. 2019. "Methionine Adenosyltransferase 1a (MAT1A) Enhances Cell Survival During Chemotherapy Treatment and is Associated with Drug Resistance in Bladder Cancer PDX Mice." International Journal of Molecular Science. 20:4983.

Parasido, Erika, Avetian, George S., Naeem, Aisha, Graham, Garrett, Pishvaian, Michael, Glasgow, Eric, Mudambi, Shaila, Lee, Yichien, Ihemelandu, Chukwuemeka, Choudhry, Muhammad, Peran, Ivana, Banerjee, Partha P., Avantaggiati, Maria, Laura, Bryant, Kirsten, Baldelli, Elisa, Pierobon, Mariaelena, Liotta, Lance, Petricoin, Emanuel, Fricke, Stanley T., Sebastian, Aimy, Cozzitorto, Joseph, Loots, Gabriela G., Kumar, Deepak, Byers, Stephen, Londin, Eric, DiFeo, Analisa, Narla, Goutham, Winter, Jordan, Brody, Jonathan R., Rodriguez, Olga and Chris Albanese. 2019. "The Sustained Induction of c-MYC Drives Nab-Paclitaxel Resistance in Primary Pancreatic Ductal Carcinoma Cells." Molecular Cancer Research 17 (9): 1815-1827.

Dubbin, Karen, Robertson, Claire, Hinckley, Aubree, Alvarado, Javier A., Gilmore, Sean F., Hynes, William F., Wheeler Elizabeth K., and Monica L. Moya. 2020. "Macromolecular gelatin properties affect fibrin microarchitecture and tumor spheroid behavior in fibrin-gelatin gels." Biomaterials. Aug;250:120035.

Hum, Nicholas R., Sebastian, Aimy, Gilmore, Sean F., He, Wei, Martin, Kelly A., Hinckley, Aubree, Dubbin, Karen R., Moya, Monica L., Wheeler, Elizabeth K., Coleman, Matthew A., and Gabriela G. Loots. 2020. "Comparative Molecular Analysis of Cancer Behavior Cultured In Vitro, In Vivo, and Ex Vivo " 2020, Cancers 12(3): 690.

Peran, Ivana, Dakshanamurthy, Sivanesan, McCoy, Matthew D., Mavropoulos, Anastasia, Allo, Bedily, Sebastian, Aimy, Hum, Nicholas R., Sprague, Sara C., Martin, Kelly A., Pishvaian, Michael J., Vietsch, Eveline E., Wellstein, Anton, Atkins, Michael B., Weiner, Louis M., Quong, Andrew A., Loots, Gabriela G., Yoo, Stephen S., Assefnia, Shahin, and Stephen W. Byers. 2020. "Cadherin 11 Promotes Immunosuppression and Extracellular Matrix Deposition to Support Growth of Pancreatic Tumors and Resistance to Gemcitabine in Mice." Gastroenterology 160, no. 4: 1359-1372.://