3D multicellular in vitro models for tumors, angiogenesis and tissue engineering

Forschungsthema

Predicting the clinical response to anti-cancer therapy is one of the major challenges in modern oncology. Despite the progress in targeted therapy approaches and comprehensive molecular sequencing techniques, many patients do not benefit from therapy and for numerous agents there are no useful predictive biomarkers to guide patient stratification. Up to now, therapeutic decision-making is based on “static” features of dead tumor tissue (histology, immunohistochemistry, mutation analysis, expression analysis) without the possibility to measure “dynamic” cellular functions or responses to specific (drug-induced) pertubation. Therefore, we aim at developing novel 3D tumor models that allow to functionally assess the response of vital tumor cells of individual patients to (targeted) therapeutic agents. The main techniques to provide clinically useful predictive information from living cancer cells include tumor tissue slice culture, organoid culture, immune cell culture, and ex vivo short-term cell cultures. The establishment of innovative tumor models will serve collaboration projects on target validation as well as research topics relying on conservation of 3D tumor complexity and heterogeneity. The long-term goal is the initiation of clinical studies to validate broadly applicable predictive biomarker tests for personalized medicine. The integration of response patterns with large-scale mutational analysis data will provide a comprehensive phenotype-genotype profile that guides individual therapeutic decisions.

Vascularization and angiogenesis are important endothelial cell functions that drive the success of wound healing, a biomaterial implant or the growth of a tumor in vivo, to name a few. An understanding of the basis of these endothelial processes is essential for determining strategies for tissue regeneration as well as for inhibiting tumor cell growth. Numerous tissue engineering approaches are being pursued in order to biofabricate a replacement for a defective tissue or organ that exhibits a correct 3-D structure and functionality after implantation. A limiting factor in the success of a biomaterial seeded with cells in vitro and biomaterial implants is a rapid delivery of blood to all cells on the implant to supply nutrients to the growing tissue. Similarly, tumor growth in vivo requires a rapidly increasing vascularization for the ever expanding tumor cell population for the same reason. Thus, in vitro primary 2- and 3-D human multi-cell culture models including endothelial cells that mimic healthy in vivo like tissue and organs as well as models with cancer cells and endothelial cells to mimic tumors are being developed in order to analyze individual cell growth and gene expression and evaluate the effects of nanoparticles, growth factors, chemotherapeutic compounds, etc. on tissue-specific cells and on vasculogenesis and angiogenesis. The specific aim is to use these tissue-like models to investigate the toxicity of biomaterials, to determine the biocompatibility of biomaterials, to identify the best physico-chemical makeup of biomaterials that allows cells to distribute and mimic the tissue, to determine conditions for the generation of a rapid vascularization of biomaterials destined for tissue regeneration and in the opposite context to evaluate the efficacy of novel chemotherapeutic approaches to inhibt the rapid vascularization of a growing tumor. The goal is to demonstrate that these in vitro models mimic the in vivo situation and can be used as tools to gain an understanding of cell-cell-material interactions, predict the success of the biomaterial and chemotherapeutic approaches and can be used as alternatives to animal studies.

Ausgewählte Publikationen

Gdynia G, Sauer SW, Kopitz J, Fuchs D, Duglova K, Ruppert T, Miller M, Pahl J, Cerwenka A, Enders M, Mairbäurl H, Kamiński MM, Penzel R, Zhang C, Fuller JC, Wade RC, Benner A, Chang-Claude J, Brenner H, Hoffmeister M, Zentgraf H, Schirmacher P, Roth W. (2016) The HMGB1 protein induces a metabolic type of tumor cell death by blocking aerobic respiration. Nature Communications, 7, 10764
Metzig MO, Fuchs D, Tagscherer KE, Gröne HJ, Schirmacher P, Roth W. (2016) Inhibition of caspases primes colon cancer cells for 5-Fluorouracil induced necroptosis driven by NF-κB. Oncogene, 35, 3399-3409
Unger RE, Dohle E, Kirkpatrick C J. (2015). Improving vascularization of engineered bone through the generation of pro-angiogenic effects in co-culture systems. Advanced Drug Delivery Reviews, 94, 116–125.
Heller M, Frerick-Ochs E V, Bauer HK, Schiegnitz E, Flesch D, Brieger J, Stein R, Al-Nawas B, Brochhausen C, Thüroff JW, Unger RE, Brenner W. (2015). Tissue engineered pre-vascularized buccal mucosa equivalents utilizing a primary triculture of epithelial cells, endothelial cells and fibroblasts. Biomaterials, 77, 207–215.
Freese C, Schreiner D, Anspach L, Bantz C, Maskos M, Unger RE, Kirkpatrick, CJ. (2014). In vitro investigation of silica nanoparticle uptake into human endothelial cells under physiological cyclic stretch. Particle and Fibre Toxicology, 11(1), 1.
Alekseeva T, Unger RE, Brochhausen C, Brown R A, Kirkpatrick JC. (2014) Engineering a microvascular capillary bed in a tissue-like collagen construct. Tissue Engineering. Part A, 20 (19-20), 2656–2665.
Weissinger D, Tagscherer KE, Macher-Goeppinger S, Haferkamp A, Wagener N, Roth W. (2013) The soluble Decoy Receptor 3 isregulated by a PI3K-dependent mechanism and promotes migration and invasion in renal cell carcinoma. Molecular Cancer 10, 120
Fassl A, Tagscherer KE, Richter J, Berriel Diaz M, Alcantara Llaguno SR, Campos B, Kopitz J, Herold-Mende C, Herzig S, Schmidt MH, Parada LF, Wiestler OD, Roth W. (2012) Notch1 signaling promotes survival of glioblastoma cells via EGFR-mediated induction of anti-apoptotic Mcl-1. Oncogene 31, 4698-708
Unger R E, Ghanaati S, Orth C, Sartoris A, Barbeck M, Halstenberg S, Motta A, Migliaresi C, Kirhpatrick CJ. (2010). The rapid anastomosis between prevascularized networks on silk fibroin scaffolds generated in vitro with cocultures of human microvascular endothelial and osteoblast cells and the host vasculature. Biomaterials, 31(27), 6959–6967.
Santos MI, Tuzlakoglu K, Fuchs S, Gomes ME, Peters K, Unger RE, Piskin E, Reis RL, Kirkpatrick JC. (2008). Endothelial cell colonization and angiogenic potential of combined nano- and micro-fibrous scaffolds for bone tissue engineering. Biomaterials, 29(32), 4306–4313.
Unger RE, Sartoris A, Peters K, Motta A, Migliaresi C, Kunkel M, Bulnheim U, Rychly J, Kirkpatrick CJ. (2007). Tissue-like self-assembly in cocultures of endothelial cells and osteoblasts and the formation of microcapillary-like structures on three-dimensional porous biomaterials. Biomaterials, 28(27), 3965–3976.