A Colorectal Cancer 3D Bioprinting Workflow as a Platform for Disease Modeling and Chemotherapeutic Screening

Abstract

Colorectal cancer (CRC) is the third most common malignancy and has recently moved up to the second leading cause of death among carcinomas. Prognosis, especially for advanced diseases or certain molecular subtypes of CRC, remains poor, which highlights the urgent need for better therapeutic strategies. However, currently, as little as 0.1% of all drugs make it from bench to bedside because of the inherently high false-positive and false-negative rates of current preclinical and clinical drug testing data. Therefore, the success of developing novel treatment agents lies in the introduction of improved preclinical disease models which resemble in vivo carcinomas closer, possess higher predictive properties, and offer opportunities for individualized therapies. Aiming to address these needs, we have established an affordable, flexible, and highly reproducible 3D bioprinted CRC model. The histological assessment of Caco-2 cells in 3D bioprints revealed the formation of glandular-like structures which show greater pathomorphological resemblance to tumors than monolayer cultures do. RNA expression profiles in 3D bioprinted cells were marked by upregulation of genes involved in cell adhesion, hypoxia, EGFR/KRAS signaling, and downregulation of cell cycle programs. Testing this 3D experimental platform with three of the most commonly used chemotherapeutics in CRC (5-fluoruracil, oxaliplatin, and irinotecan) revealed overall increased resistance compared to 2D cell cultures. Last, we demonstrate that our workflow can be successfully extended to primary CRC samples. Thereby, we describe a novel accessible platform for disease modeling and drug testing, which may present an innovative opportunity for personalized therapeutic screening.

Characterisation of 3D Bioprinted Human Breast Cancer Model for In Vitro Drug and Metabolic Targeting

“…[65,[68][69][70][71][72]. Modelling the complexity of tumour progression, especially the steps of metastasis formation, also requires microfluidics and several host tissues to be combined in novel in vitro test systems [82][83][84][85][86][87][88][89][90][91][92][93]. The continuously emerging data about such devices for BC and more complex BC models were summarised by Moccia and Haase [94,95].…”

Constructing 3D In Vitro Models of Heterocellular Solid Tumors and Stromal Tissues Using Extrusion-Based Bioprinting

“…Interestingly, they also found an upregulation of HIF-1α, which regulates vascularization of in the TME. 64 Other cancer types recapitulated in vitro as spheroids inside bioprinted matrices include breast cancer, 65 melanoma, 66 colorectal cancer, 67 lung cancer, 68 glioma, 69 glioblastoma, 63 neuroblastoma, 70 and cervical cancer. 71 A key factor in tumor sphere elucidation for drug testing purposes is maintaining spheroid size uniformity.…”

The potential role of the three-dimensional-bioprinting model in screening and developing drugs

“…However, 3D-bioprinting provides several critical advantages over sphere formation assay in drug development or screening, such as using bio-ink to simulate the cytoskeleton or partial tumor tissue with multi-cell to a highly complex hierarchical 3D structure. These configurational were able to enhance intercellular communication and signaling factors transportation and provide a more accurate result for novel drug development[ 4 , 5 ]. Although the authors used organoid cultures to verify the drug's effectiveness at a later stage, the success rate of organ-like laboratory cultures is too low, which significantly limits the possibility of large-scale experimental validation.…”