title: High-throughput screening with nanoimprinting 3D culture for efficient drug development by mimicking the tumor environment.
abstract: Anti-cancer drug development typically utilizes high-throughput screening with two-dimensional (2D) cell culture. However, 2D culture induces cellular characteristics different from tumors in vivo, resulting in inefficient drug development. Here, we report an innovative high-throughput screening system using nanoimprinting 3D culture to simulate in vivo conditions, thereby facilitating efficient drug development. We demonstrated that cell line-based nanoimprinting 3D screening can more efficiently select drugs that effectively inhibit cancer growth in vivo as compared to 2D culture. Metabolic responses after treatment were assessed using positron emission tomography (PET) probes, and revealed similar characteristics between the 3D spheroids and in vivo tumors. Further, we developed an advanced method to adopt cancer cells from patient tumor tissues for high-throughput drug screening with nanoimprinting 3D culture, which we termed Cancer tissue-Originated Uniformed Spheroid Assay (COUSA). This system identified drugs that were effective in xenografts of the original patient tumors. Nanoimprinting 3D spheroids showed low permeability and formation of hypoxic regions inside, similar to in vivo tumors. Collectively, the nanoimprinting 3D culture provides easy-handling high-throughput drug screening system, which allows for efficient drug development by mimicking the tumor environment. The COUSA system could be a useful platform for drug development with patient cancer cells.
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan. Electronic address: yukiey@nirs.go.jp.
authors.forename: Yukie
authors.initials: Y
authors.lastname: Yoshii
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Furukawa
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan.
authors.forename: Atsuo
authors.initials: A
authors.lastname: Waki
authors.affiliation: Department of Biochemistry, Osaka Medical Center for Cancer and Cardiovascular Diseases, Higashinari-ku, Osaka 537-8511, Japan.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Okuyama
authors.affiliation: Department of Biochemistry, Osaka Medical Center for Cancer and Cardiovascular Diseases, Higashinari-ku, Osaka 537-8511, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Inoue
authors.affiliation: SCIVAX Life Sciences, Inc., Kawasaki, Kanagawa 213-0012, Japan.
authors.forename: Manabu
authors.initials: M
authors.lastname: Itoh
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan.
authors.forename: Ming-Rong
authors.initials: MR
authors.lastname: Zhang
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan.
authors.forename: Hidekatsu
authors.initials: H
authors.lastname: Wakizaka
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan; The Jackson Laboratory, Bar Harbor, ME 04609, USA.
authors.forename: Chizuru
authors.initials: C
authors.lastname: Sogawa
authors.affiliation: Biomedical Imaging Research Center, University of Fukui, Eiheiji, Fukui 910-1193, Japan.
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Kiyono
authors.affiliation: Research Center for Radiation Emergency Medicine, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Yoshii
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan.
authors.forename: Yasuhisa
authors.initials: Y
authors.lastname: Fujibayashi
authors.affiliation: Molecular Imaging Center, National Institute of Radiological Sciences, Anagawa, Chiba 263-8555, Japan.
authors.forename: Tsuneo
authors.initials: T
authors.lastname: Saga
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001681:Biological Assay; D018929:Cell Culture Techniques / Q000379:methods*; D015687:Cell Hypoxia / Q000187:drug effects; D045744:Cell Line, Tumor; D015195:Drug Design; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D036103:Nanotechnology / Q000379:methods*; D009369:Neoplasms / Q000473:pathology; D018874:Spheroids, Cellular / Q000187:drug effects / Q000378:metabolism; D059016:Tumor Microenvironment / Q000187:drug effects
chemical_list: D000970:Antineoplastic Agents; D007976:Leupeptins; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde
keywords: 3D cell culture; High-throughput drug screening; Hypoxia; Nanoimprinting technology; Spheroid