High‐throughput screening in colorectal cancer tissue‐originated spheroids

Patient‐derived cancer organoid culture is an important live material that reflects clinical heterogeneity. However, the limited amount of organoids available for each case as well as the considerable amount of time and cost to expand in vitro makes it impractical to perform high‐throughput drug screening using organoid cultures from multiple patients. Here, we report an advanced system for the high‐throughput screening of 2427 drugs using the cancer tissue‐originated spheroid (CTOS) method. In this system, we apply the CTOS method in an ex vivo platform from xenograft tumors, using machines to handle CTOS and reagents, and testing a CTOS reference panel of multiple CTOS lines for the hit drugs. CTOS passages in xenograft tumors resulted in minimal changes of morphological and genomic status, and xenograft tumor generation efficiently expanded the number of CTOS to evaluate multiple drugs. Our panel of colorectal cancer CTOS lines exhibited diverse sensitivities to the hit compounds, demonstrating the usefulness of this system for investigating highly heterogeneous disease.

[1]  H. Clevers,et al.  Organoids in cancer research , 2018, Nature Reviews Cancer.

[2]  Andrea Sottoriva,et al.  Patient-derived organoids model treatment response of metastatic gastrointestinal cancers , 2018, Science.

[3]  C. Dejong,et al.  Patient‐derived organoid models help define personalized management of gastrointestinal cancer , 2018, The British journal of surgery.

[4]  A. Letai,et al.  Functional precision cancer medicine—moving beyond pure genomics , 2017, Nature Medicine.

[5]  Rameen Beroukhim,et al.  Patient-derived xenografts undergo murine-specific tumor evolution , 2017, Nature Genetics.

[6]  M. Inoue,et al.  The induction of MIG6 under hypoxic conditions is critical for dormancy in primary cultured lung cancer cells with activating EGFR mutations , 2017, Oncogene.

[7]  Davide Prandi,et al.  Personalized In Vitro and In Vivo Cancer Models to Guide Precision Medicine. , 2017, Cancer discovery.

[8]  M. Ohue,et al.  In vivo and ex vivo cetuximab sensitivity assay using three-dimensional primary culture system to stratify KRAS mutant colorectal cancer , 2017, PloS one.

[9]  Catherine L. Worth,et al.  Molecular dissection of colorectal cancer in pre-clinical models identifies biomarkers predicting sensitivity to EGFR inhibitors , 2017, Nature Communications.

[10]  Marc Bickle,et al.  Screening out irrelevant cell-based models of disease , 2016, Nature Reviews Drug Discovery.

[11]  Gordon B Mills,et al.  Integrated Patient-Derived Models Delineate Individualized Therapeutic Vulnerabilities of Pancreatic Cancer. , 2016, Cell reports.

[12]  M. Inoue,et al.  Drug screening and grouping by sensitivity with a panel of primary cultured cancer spheroids derived from endometrial cancer , 2016, Cancer science.

[13]  M. Hiraoka,et al.  Radiation sensitivity assay with a panel of patient‐derived spheroids of small cell carcinoma of the cervix , 2015, International journal of cancer.

[14]  Hayley E. Francies,et al.  Prospective Derivation of a Living Organoid Biobank of Colorectal Cancer Patients , 2015, Cell.

[15]  Ming-Rong Zhang,et al.  High-throughput screening with nanoimprinting 3D culture for efficient drug development by mimicking the tumor environment. , 2015, Biomaterials.

[16]  N. Nonomura,et al.  High-dose chemotherapeutics of intravesical chemotherapy rapidly induce mitochondrial dysfunction in bladder cancer-derived spheroids , 2014, Cancer science.

[17]  Hans Clevers,et al.  Organoid cultures for the analysis of cancer phenotypes. , 2014, Current opinion in genetics & development.

[18]  B. Giusti,et al.  EXCAVATOR: detecting copy number variants from whole-exome sequencing data , 2013, Genome Biology.

[19]  A. Sivachenko,et al.  Sensitive detection of somatic point mutations in impure and heterogeneous cancer samples , 2013, Nature Biotechnology.

[20]  M. Higashiyama,et al.  Spheroid Culture of Primary Lung Cancer Cells with Neuregulin 1/HER3 Pathway Activation , 2013, Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer.

[21]  Christopher A. Miller,et al.  VarScan 2: somatic mutation and copy number alteration discovery in cancer by exome sequencing. , 2012, Genome research.

[22]  H. Iishi,et al.  Retaining cell–cell contact enables preparation and culture of spheroids composed of pure primary cancer cells from colorectal cancer , 2011, Proceedings of the National Academy of Sciences.

[23]  M. DePristo,et al.  A framework for variation discovery and genotyping using next-generation DNA sequencing data , 2011, Nature Genetics.

[24]  R. Shoemaker.  The NCI60 human tumour cell line anticancer drug screen , 2006, Nature Reviews Cancer.

[25]  T. Hongyo,et al.  Effects of N-methyl-N'-nitro-N-nitrosoguanidine on the human colorectal polyps consecutively maintained in SCID mice. , 2002, Cancer letters.

[26]  T. Tsuruo,et al.  Potent antitumor activity of MS-247, a novel DNA minor groove binder, evaluated by an in vitro and in vivo human cancer cell line panel. , 1999, Cancer research.

[27]  D. Scudiero,et al.  Feasibility of a high-flux anticancer drug screen using a diverse panel of cultured human tumor cell lines. , 1991, Journal of the National Cancer Institute.

[28]  Juergen Friedrich,et al.  Spheroid-based drug screen: considerations and practical approach , 2009, Nature Protocols.

[29]  Claude-Alain H. Roten,et al.  Fast and accurate short read alignment with Burrows–Wheeler transform , 2009, Bioinform..