title: Novel Morphologic and Genetic Analysis of Cancer Cells in a 3D Microenvironment Identifies STAT3 as a Regulator of Tumor Permeability Barrier Function.
abstract: Tumor permeability is a critical determinant of drug delivery and sensitivity, but systematic methods to identify factors that perform permeability barrier functions in the tumor microenvironment are not yet available. Multicellular tumor spheroids have become tractable in vitro models to study the impact of a three-dimensional (3D) environment on cellular behavior. In this study, we characterized the spheroid-forming potential of cancer cells and correlated the resulting spheroid morphologies with genetic information to identify conserved cellular processes associated with spheroid structure. Spheroids generated from 100 different cancer cell lines were classified into four distinct groups based on morphology. In particular, round and compact spheroids exhibited highly hypoxic inner cores and permeability barriers against anticancer drugs. Through systematic and correlative analysis, we reveal JAK-STAT signaling as one of the signature pathways activated in round spheroids. Accordingly, STAT3 inhibition in spheroids generated from the established cancer cells and primary glioblastoma patient-derived cells altered the rounded morphology and increased drug sensitivity. Furthermore, combined administration of the STAT3 inhibitor and 5-fluorouracil to a mouse xenograft model markedly reduced tumor growth compared with monotherapy. Collectively, our findings demonstrate the ability to integrate 3D culture and genetic profiling to determine the factors underlying the integrity of the permeability barrier in the tumor microenvironment, and may help to identify and exploit novel mechanisms of drug resistance.
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea. Advanced Institutes of Convergence Technology, Seoul National University, Suwon, Korea.
authors.forename: Min Chul
authors.initials: MC
authors.lastname: Park
authors.affiliation: School of Interdisciplinary Bioscience and Bioengineering, POSTECH, Pohang, Korea.
authors.forename: Hyobin
authors.initials: H
authors.lastname: Jeong
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea.
authors.forename: Sung Hwa
authors.initials: SH
authors.lastname: Son
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea. Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Technology, College of Pharmacy, Seoul National University, Seoul, Korea.
authors.forename: YounHa
authors.initials: Y
authors.lastname: Kim
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea. Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Technology, College of Pharmacy, Seoul National University, Seoul, Korea.
authors.forename: Daeyoung
authors.initials: D
authors.lastname: Han
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea. Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Technology, College of Pharmacy, Seoul National University, Seoul, Korea.
authors.forename: Peter C
authors.initials: PC
authors.lastname: Goughnour
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea.
authors.forename: Taehee
authors.initials: T
authors.lastname: Kang
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea.
authors.forename: Nam Hoon
authors.initials: NH
authors.lastname: Kwon
authors.affiliation: Department of Neurosurgery, Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Hyo Eun
authors.initials: HE
authors.lastname: Moon
authors.affiliation: Department of Neurosurgery, Cancer Research Institute, Seoul National University College of Medicine, Seoul, Korea.
authors.forename: Sun Ha
authors.initials: SH
authors.lastname: Paek
authors.affiliation: School of Interdisciplinary Bioscience and Bioengineering, POSTECH, Pohang, Korea. Center for Systems Biology of Plant Senescence and Life History, Institute for Basic Science, Department of New Biology, DGIST, Daegu, Korea.
authors.forename: Daehee
authors.initials: D
authors.lastname: Hwang
authors.affiliation: Department of Neurosurgery, Samsung Medical Center, School of Medicine, Sungkyunkwan University, Seoul, Korea.
authors.forename: Ho Jun
authors.initials: HJ
authors.lastname: Seol
authors.affiliation: Department of Neurosurgery, Samsung Medical Center, School of Medicine, Sungkyunkwan University, Seoul, Korea.
authors.forename: Do-Hyun
authors.initials: DH
authors.lastname: Nam
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul, Korea. Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Technology, College of Pharmacy, Seoul National University, Seoul, Korea. The National Center for Drug Screening, Shanghai Institute of Materia Medica, Chinese Academy of Sciences, Shanghai, China. sungkim@snu.ac.kr.
authors.forename: Sunghoon
authors.initials: S
authors.lastname: Kim
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000494:pharmacology; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D019008:Drug Resistance, Neoplasm; D005472:Fluorouracil / Q000494:pharmacology; D006801:Humans; D053612:Janus Kinases / Q000502:physiology; D047029:Lactams, Macrocyclic / Q000494:pharmacology; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D050796:STAT3 Transcription Factor / Q000502:physiology*; D015398:Signal Transduction; D018874:Spheroids, Cellular; D059016:Tumor Microenvironment; D020032:Tyrphostins / Q000494:pharmacology
chemical_list: D016227:Benzoquinones; D047029:Lactams, Macrocyclic; D050796:STAT3 Transcription Factor; C494086:STAT3 protein, human; D020032:Tyrphostins; C095512:alpha-cyano-(3,4-dihydroxy)-N-benzylcinnamide; C112765:tanespimycin; D053612:Janus Kinases; D005472:Fluorouracil