Abstract 2691: 4D printing of programmable smart material for drug screening in patient-derived organoids
Authors: Yang Chen , Chadwick Michelle , Lee Howon , Sabaawy Hatem E. 
Published: August 2019

Abstract
Numerous studies have contributed to a paradigm shift in cancer modeling, moving from traditional two-dimensional (2D) culture to three-dimensional (3D) sphere and organoid cultures, to recapitulate key aspects of in vivo multicellular, anatomical and functional organ complexity. In parallel, biomaterial 3D printing advances allowed generating devices, implants, and scaffolds for tissue engineering and drug delivery systems. Despite these advances, platforms that integrate genomic, proteomic, and bioprinting innovations for high-throughput screening (HTS) or selection of effective personalized cancer treatments are scarce. Here, we utilized cell-based 3D organoids with four-dimensional (4D) printing of smart biomaterial to generate devices and platforms for personalized and HTS of drugs. Because genomic assays alone are insufficient to identify therapeutic options for most patients, we integrated genomic sequencing, patient derived organoids (PDOs), histological and immunohistochemical (IHC) assessments with evaluating tumor cell responses to therapies. First, we developed a novel 4D printed temperature-sensitive smart material consisting of interconnected basket arrays that fit into 96-well plates. The temperature-responsive 4D-printed basket arrays were fabricated using projection micro-stereolithography (PμSL), with a spatial light modulator used as a reconfigurable photomask, and projection through reduction lenses into photo curable shape polymer resin. These 4D-printed baskets are 3.4 mm-deep and have a 1.8 mm-opening, into which cells, media and matrices could be plated (manually or automated) to generate 3D spheres and organoids, to be treated with different drugs. We have utilized these 4D-printed basket arrays to generate PDOs from glioblastoma (GBM) tissues that were subjected to exome sequencing. Single cell derived GBM were allowed to form PDOs for 2 weeks, followed by 72-hour treatments with various compounds, such as temozolomide, rapamycin, PI3K and BMI-1 inhibitors, among others. Following treatments, the entire 4D printed basket arrays were evolved, with a 10-min heating step at 50ºC, to their programmable cassette size to directly perform histological and IHC validation on the same day. PDOs were examined for cell proliferation by quantifying intracellular adenosine triphosphate (ATP), tumor invasiveness and pathway-specific markers to assess drug efficacy. This novel platform allowed the entire patient tissue and drug response assessment to be completed in ≤20 days. When including exome and/or single cell sequencing, histological, IHC and targeted therapeutic assays, our basket array platform could offer dynamic, automated, and quantitative drug analyses, thus allow the discovery of novel preclinical therapeutic approaches that can be assessed in clinical trials, and provides personalized therapies for individual patients in precision medicine.
Citation Format: Chen Yang, Michelle Chadwick, Howon Lee, Hatem E. Sabaawy. 4D printing of programmable smart material for drug screening in patient-derived organoids [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 2691.