Application of 3D tumoroid systems to define immune and cytotoxic therapeutic responses based on tumoroid and tissue slice culture molecular signatures

Abstract

We have developed 3D-tumoroids and tumor slice in vitro culture systems from surgical tumor specimens derived from patients with colorectal cancer (CRC) or lung cancer to evaluate immune cell populations infiltrating cultured tissues. The system incorporates patient's peripherally and tumor-derived immune cells into tumoroid in vitro cultures to evaluate the ability of the culture to mimic an immunosuppressive tumor microenvironment (ITM). This system enables analysis of tumor response to standard therapy within weeks of surgical resection. Here we show that tumoroid cultures from a CRC patient are highly sensitive to the thymidylate synthase inhibitor 5-fluorouracil (adrucil) but less sensitive to the combination of nucleoside analog trifluridine and thymidine phosphorylase inhibitor tipiracil (Lonsurf). Moreover, re-introduction of isolated immune cells derived from surrounding and infiltrating tumor tissue as well as CD45+ tumor infiltrating hematopoietic cells displayed prolonged (>10 days) survival in co-culture. Established tumor slice cultures were found to contain both an outer epithelial and inner stromal cell compartment mimicking tumor structure in vivo.Collectively, these data suggest that, 3D-tumoroid and slice culture assays may provide a feasible in vitro approach to assess efficacy of novel therapeutics in the context of heterogeneous tumor-associated cell types including immune and non-transformed stromal cells. In addition, delineating the impact of therapeutics on immune cells, and cell types involved in therapeutic resistance mechanisms may be possible in general or for patient-specific responses.

Three-Dimensional in Vitro Cell Culture Models in Drug Discovery and Drug Repositioning

“…For example, comparison of drug response profiles of breast, prostate and lung cancer cell lines revealed clear differences in dose response curves to docetaxel and fulvestrant when cells were grown in collagen and compared to 2D cultures or other 3D culture systems such as low attachment plates and other natural scaffolds, including alginate and Matrigel (Stock et al, 2016 ). Multiple myeloma cells cultured in a Matrigel-based human bone marrow-like microenvironment provide a system for preclinical testing of chemotherapeutics that take into account adhesion-mediated mechanisms of drug resistance (Kirshner et al, 2008 ) and Matrigel-embedded 3D-tumoroids derived from tissue of patients with colorectal cancer and lung cancer can provide a 3D culture system for drug testing that contains not only tumor cells but also immune cells from surrounding tissues (Finnberg et al, 2017 ). Matrigel and similar products are a gelatinous mixture of proteins and growth factors secreted by Engelbreth-Holm-Swarm mouse sarcoma cells (Kleinman and Martin, 2005 ).…”

Modeling neoplastic disease with spheroids and organoids

“…Organoids resemble the heterogeneous cytoarchitecture found in vivo and advanced microscopy techniques can be used to follow the dynamic processes of organoid development and maturation been made in cancer immunotherapy, and several organoid-based models have been created to better our understanding on how the immune system eradicates tumor cells, facilitating a personalized immunotherapeutic approach [171]. Currently, two conceptually different organoid models are used-i.e., cancer organoids cultured directly from tumors preserving endogenous immune cells and other non-epithelial cell types (holistic approach) [172][173][174][175], and cancer organoids co-cultured with immune cell subsets isolated and separately expanded (reductionist approach) [176][177][178]. The two are equally valid approaches to understanding cell-cell interactions, modelling immune checkpoint blockade and testing CAR-T cell-mediated cytotoxicity and represent a highly informative platform for the development of cancer immunotherapy.…”

Organoid Models of Colorectal Pathology: Do They Hold the Key to Personalized Medicine? A Systematic Review

“… 70 – 74 Tissue-derived organoids can be cocultured with local tumor-infiltrating immune cells to assess the response to chemotherapeutic agents and radiation. 75 Along with the ability to assess the effects of chemotherapeutic agents and radiation directly on patient tumor samples, organoids also allow us to more deeply analyze the transcriptional and proteomic profiles of individual patients. Using normal and cancer organoids derived from the same patient, Cristobal and coworkers 76 determined a CRC-specific proteomic profile that showed a significant enrichment in Wnt signaling as would be expected, along with unique tumor proteomic signatures.…”