Han, Jeonghun Jun, Yukyung Kim, So Hyun Hoang, Hong-Hoa Jung, Yeonjoo Kim, Suyeon Kim, Jaesang Austin, Robert H. Lee, Sanghyuk Park, Sungsu 2024-01-20T02:34:04Z 2024-01-20T02:34:04Z 2022-01-25 2016-12 0027-8424 https://pubs.kist.re.kr/handle/201004/123339 In vitro prediction of the probable rapid emergence of resistance to a drug in tumors could act to winnow out potential candidates for further costly development. We have developed a microfluidic device consisting of similar to 500 hexagonal microcompartments that provides a complex ecology with wide ranges of drug and nutrient gradients and local populations. This ecology of a fragmented metapopulation induced the drug resistance in stage IV U87 glioblastoma cells to doxorubicin in 7 d. Exome and transcriptome sequencing of the resistant cells identified mutations and differentially expressed genes. Gene ontology and pathway analyses of the genes identified showed that they were functionally relevant to the established mechanisms of doxorubicin action. Specifically, we identified (i) a frame-shift insertion in the filamin-A gene, which regulates the influx and efflux of topoisomerase II poisons; (ii) the overexpression of aldo-keto reductase enzymes, which convert doxorubicin into doxorubicinol; and (iii) activation of NF-kappa B via alterations in the nucleotide-binding oligomerization domain (NOD)-like receptor signaling pathway from mutations in three genes (CARD6, NSD1, and NLRP13) and the overexpression of inflammatory cytokines. Functional experiments support the in silico analyses and, together, demonstrate the effects of these genetic changes. Our findings suggest that, given the rapid evolution of resistance and the focused response, this technology could act as a rapid screening modality for genetic aberrations leading to resistance to chemotherapy as well as counter selection of drugs unlikely to be successful ultimately. English NATL ACAD SCIENCES Rapid emergence and mechanisms of resistance by U87 glioblastoma cells to doxorubicin in an in vitro tumor microfluidic ecology Article 10.1073/pnas.1614898113 1 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, v.113, no.50, pp.14283 - 14288 PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 113 50 14283 14288 N scie scopus 000389696700049 Multidisciplinary Sciences Science & Technology - Other Topics Article NF-KAPPA-B CANCER DRUG-RESISTANCE ANALYSIS TOOLKIT ACTIVATION MUTATIONS PATHWAYS CULTURE GENES microhabitats cancer doxorubicin evolution resistance