Novel bioassays based on 3D-printed device for sensing of hypoxia and p53 pathway in 3D cell models.
3D cell model
3D-printed device
Bioactivity
Bioluminescence
Hypoxia
p53
Journal
Analytical and bioanalytical chemistry
ISSN: 1618-2650
Titre abrégé: Anal Bioanal Chem
Pays: Germany
ID NLM: 101134327
Informations de publication
Date de publication:
19 Oct 2024
19 Oct 2024
Historique:
received:
02
08
2024
accepted:
08
10
2024
revised:
29
09
2024
medline:
19
10
2024
pubmed:
19
10
2024
entrez:
19
10
2024
Statut:
aheadofprint
Résumé
Cell-based assays are widely exploited for drug screening and biosensing, providing useful information about bioactivity of target analytes and complex biological samples. It is well recognized that 3D cell models are required to achieve highly valuable information, also from the perspective of replacing animal models. However, bioassays relying on 3D cell models are generally highly demanding in terms of facilities, equipment, and skilled personnel requirements. To reduce cost, increase sustainability, and provide a flexible 3D cell-based platform for bioassays, we here report a novel approach based on a 3D-printed microtissue device. To assess the suitability of this strategy for reporter gene technology, we selected to monitor two molecular pathways which were of interest in several applications, hypoxia signaling and the p53 pathway. The investigation of such pathways is highly relevant in fields spanning from drug screening to bioactivity monitoring for industrial by-product valorization. Microtissues of human hepatocarcinoma (HepG2) and human embryonic kidney (Hek293T) cell lines were obtained with a low-cost and sustainable chip platform and bioassays were developed to monitor the hypoxia-inducible factors (HIFs) and the p53 tumor suppressor pathway. HepG2 and Hek293T 3D cell models were genetically engineered to express the Luc2P from Photinus pyralis firefly either under the regulation of p53 or HIF response elements. The bioassays allowed quantitative assessment of hypoxia and tumoral activity with 1,10-phenanthroline for HIF and with doxorubicin for p53 pathway activation, respectively, showing good potential for applications of this sustainable and low-cost 3D-printed microfluidic platform for bioactivity analyses, drug screening, and precision medicine.
Identifiants
pubmed: 39425762
doi: 10.1007/s00216-024-05606-0
pii: 10.1007/s00216-024-05606-0
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Subventions
Organisme : European Commission
ID : 0001052
Organisme : European Commission
ID : CN00000022
Organisme : HORIZON EUROPE Food, Bioeconomy, Natural Resources, Agriculture and Environment
ID : 101135533
Informations de copyright
© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.
Références
Russell WMS, Burch RL (1960) The principles of humane experimental technique. Med J Australia 1:500–500. https://doi.org/10.5694/j.1326-5377.1960.tb73127.x.
Calabretta MM, Michelini E. Current advances in the use of bioluminescence assays for drug discovery: an update of the last ten years. Expert Opin Drug Discov. 2024;19:85–95. https://doi.org/10.1080/17460441.2023.2266989 .
doi: 10.1080/17460441.2023.2266989
pubmed: 37814480
Lenin S, Ponthier E, Scheer KG, Yeo ECF, Tea MN, Ebert LM, Oksdath Mansilla M, Poonnoose S, Baumgartner U, Day BW, Ormsby RJ, Pitson SM, Gomez GA. A drug screening pipeline using 2D and 3D patient-derived in vitro models for pre-clinical analysis of therapy response in glioblastoma. Int J Mol Sci. 2021;22:4322. https://doi.org/10.3390/ijms22094322 .
doi: 10.3390/ijms22094322
pubmed: 33919246
pmcid: 8122466
Prince E, Kheiri S, Wang Y, Xu F, Cruickshank J, Topolskaia V, Tao H, Young EWK, AlisonP McGuigan, Cescon DW, Kumacheva E. Microfluidic arrays of breast tumor spheroids for drug screening and personalized cancer therapies. Adv Healthc Mater. 2022;11:e2101085. https://doi.org/10.1002/adhm.202101085 .
doi: 10.1002/adhm.202101085
pubmed: 34636180
Żuchowska A, Baranowska P, Flont M, Brzózka Z, Jastrzębska E. Review: 3D cell models for organ-on-a-chip applications. Anal Chim Acta. 2024;1301: 342413. https://doi.org/10.1016/j.aca.2024.342413 .
doi: 10.1016/j.aca.2024.342413
pubmed: 38553129
Totaro G, Sisti L, Vannini M, Marchese P, Tassoni A, Lenucci MS, Lamborghini M, Kalia S, Celli A. A new route of valorization of rice endosperm by-product: production of polymeric biocomposites. Compos B Eng. 2018;139:195–202. https://doi.org/10.1016/j.compositesb.2017.11.055 .
doi: 10.1016/j.compositesb.2017.11.055
Asma ST, Acaroz U, Imre K, Morar A, Shah SRA, Hussain SZ, Arslan-Acaroz D, Demirbas H, Hajrulai-Musliu Z, Istanbullugil FR, Soleimanzadeh A, Morozov D, Zhu K, Herman V, Ayad A, Athanassiou C, Ince S. Natural products/bioactive compounds as a source of anticancer drugs. Cancers (Basel). 2022;14:6203. https://doi.org/10.3390/cancers14246203 .
doi: 10.3390/cancers14246203
pubmed: 36551687
Mármol I, Quero J, Ibarz R, Ferreira-Santos P, Teixeira JA, Rocha CMR, Pérez-Fernández M, García-Juiz S, Osada J, Martín-Belloso O, Rodríguez-Yoldi MJ. Valorization of agro-food by-products and their potential therapeutic applications. Food Bioprod Process. 2021;128:247–58. https://doi.org/10.1016/j.fbp.2021.06.003 .
doi: 10.1016/j.fbp.2021.06.003
Socas-Rodríguez B, Álvarez-Rivera G, Valdés A, Ibáñez E, Cifuentes A. Food by-products and food wastes: are they safe enough for their valorization? Trends Food Sci Technol. 2021;114:133–47. https://doi.org/10.1016/j.tifs.2021.05.002 .
doi: 10.1016/j.tifs.2021.05.002
Mohammadnezhad P, Valdés A, Álvarez-Rivera G. Bioactivity of food by-products: an updated insight. Curr Opin Food Sci. 2023;52: 101065. https://doi.org/10.1016/j.cofs.2023.101065 .
doi: 10.1016/j.cofs.2023.101065
Rather RA, Bhagat M. Quercetin as an innovative therapeutic tool for cancer chemoprevention: molecular mechanisms and implications in human health. Cancer Med. 2020;9:9181–92. https://doi.org/10.1002/cam4.1411 .
doi: 10.1002/cam4.1411
pubmed: 31568659
Nagle D, Zhou Y-D. Natural product-based inhibitors of hypoxia-inducible factor-1 (HIF-1). Curr Drug Targets. 2006;7:355–69. https://doi.org/10.2174/138945006776054979 .
doi: 10.2174/138945006776054979
pubmed: 16515532
pmcid: 2908043
Zhang C, Liu J, Wang J, Zhang T, Xu D, Hu W, Feng Z. The interplay between tumor suppressor p53 and hypoxia signaling pathways in cancer. Front Cell Dev Biol. 2021;9:648808. https://doi.org/10.3389/fcell.2021.648808 .
doi: 10.3389/fcell.2021.648808
pubmed: 33681231
pmcid: 7930565
Wicks EE, Semenza GL. Hypoxia-inducible factors: cancer progression and clinical translation. J Clin Invest. 2022;132:e159839. https://doi.org/10.1172/JCI159839 .
doi: 10.1172/JCI159839
pubmed: 35642641
pmcid: 9151701
Petrova V, Annicchiarico-Petruzzelli M, Melino G, Amelio I. The hypoxic tumour microenvironment. Oncogenesis. 2018;7:10. https://doi.org/10.1038/s41389-017-0011-9 .
doi: 10.1038/s41389-017-0011-9
pubmed: 29362402
pmcid: 5833859
Choueiri TK, Kaelin WG. Targeting the HIF2–VEGF axis in renal cell carcinoma. Nat Med. 2020;26:1519–30. https://doi.org/10.1038/s41591-020-1093-z .
doi: 10.1038/s41591-020-1093-z
pubmed: 33020645
Zhang C, Liu J, Xu D, Zhang T, Hu W, Feng Z. Gain-of-function mutant p53 in cancer progression and therapy. J Mol Cell Biol. 2020;12:674–87. https://doi.org/10.1093/jmcb/mjaa040 .
doi: 10.1093/jmcb/mjaa040
pubmed: 32722796
pmcid: 7749743
Hsu C-W, Huang R, Khuc T, Shou D, Bullock J, Grooby S, Griffin S, Zou C, Little A, Astley H, Xia M. Identification of approved and investigational drugs that inhibit hypoxia-inducible factor-1 signaling. Oncotarget. 2016;7:8172–83. https://doi.org/10.18632/oncotarget.6995 .
doi: 10.18632/oncotarget.6995
pubmed: 26882567
pmcid: 4884984
Aggarwal V, Miranda O, Johnston PA, Sant S. Three dimensional engineered models to study hypoxia biology in breast cancer. Cancer Lett. 2020;490:124–42. https://doi.org/10.1016/j.canlet.2020.05.030 .
doi: 10.1016/j.canlet.2020.05.030
pubmed: 32569616
pmcid: 7442747
Cevenini L, Calabretta MM, Lopreside A, Tarantino G, Tassoni A, Ferri M, Roda A, Michelini E. Exploiting NanoLuc luciferase for smartphone-based bioluminescence cell biosensor for (anti)-inflammatory activity and toxicity. Anal Bioanal Chem. 2016;408:8859–68. https://doi.org/10.1007/s00216-016-0062-3 .
doi: 10.1007/s00216-016-0062-3
pubmed: 27853830
Lopreside A, Calabretta MM, Montali L, Ferri M, Tassoni A, Branchini BR, Southworth T, D’Elia M, Roda A, Michelini E. Prêt-à-porter nanoYESα and nanoYESβ bioluminescent cell biosensors for ultrarapid and sensitive screening of endocrine-disrupting chemicals. Anal Bioanal Chem. 2019;411:4937–49. https://doi.org/10.1007/s00216-019-01805-2 .
doi: 10.1007/s00216-019-01805-2
pubmed: 30972468
Kirchherr J, Reike D, Hekkert M. Conceptualizing the circular economy: an analysis of 114 definitions. Resour Conserv Recycl. 2017;127:221–32. https://doi.org/10.1016/j.resconrec.2017.09.005 .
doi: 10.1016/j.resconrec.2017.09.005
Cevenini L, Calabretta MM, Calabria D, Roda A, Michelini E (2015) Luciferase genes as reporter reactions: how to use them in molecular biology? pp 3–17.
D’Alessandro S, Camarda G, Corbett Y, Siciliano G, Parapini S, Cevenini L, Michelini E, Roda A, Leroy D, Taramelli D, Alano P. A chemical susceptibility profile of the Plasmodium falciparum transmission stages by complementary cell-based gametocyte assays. J Antimicrob Chemother. 2016;71:1148–58. https://doi.org/10.1093/jac/dkv493 .
doi: 10.1093/jac/dkv493
pubmed: 26888912
Lopreside A, Montali L, Wang B, Tassoni A, Ferri M, Calabretta MM, Michelini E. Orthogonal paper biosensor for mercury(II) combining bioluminescence and colorimetric smartphone detection. Biosens Bioelectron. 2021;194:113569. https://doi.org/10.1016/j.bios.2021.113569 .
doi: 10.1016/j.bios.2021.113569
pubmed: 34438340
Calabretta MM, Gregucci D, Michelini E. New synthetic red- and orange-emitting luciferases to upgrade in vitro and 3D cell biosensing. Analyst. 2023;148:5642–9. https://doi.org/10.1039/D3AN01251D .
doi: 10.1039/D3AN01251D
pubmed: 37791570
Calabretta M, Gregucci D, Martínez-Pérez-Cejuela H, Michelini E. A luciferase mutant with improved brightness and stability for whole-cell bioluminescent biosensors and in vitro biosensing. Biosensors (Basel). 2022;12:742. https://doi.org/10.3390/bios12090742 .
doi: 10.3390/bios12090742
pubmed: 36140127
Calabretta MM, Gregucci D, Guardigli M, Michelini E. Low-cost and sustainable smartphone-based tissue-on-chip device for bioluminescence biosensing. Biosens Bioelectron. 2024;261:116454. https://doi.org/10.1016/j.bios.2024.116454 .
doi: 10.1016/j.bios.2024.116454
pubmed: 38875866
Wang Y, Gao Y, Pan Y, Zhou D, Liu Y, Yin Y, Yang J, Wang Y, Song Y. Emerging trends in organ-on-a-chip systems for drug screening. Acta Pharm Sin B. 2023;13:2483–509. https://doi.org/10.1016/j.apsb.2023.02.006 .
doi: 10.1016/j.apsb.2023.02.006
pubmed: 37425038
pmcid: 10326261
Calabretta MM, Lopreside A, Montali L, Zangheri M, Evangelisti L, D’Elia M, Michelini E. Portable light detectors for bioluminescence biosensing applications: a comprehensive review from the analytical chemist’s perspective. Anal Chim Acta. 2022;1200:339583. https://doi.org/10.1016/j.aca.2022.339583 .
doi: 10.1016/j.aca.2022.339583
pubmed: 35256132
McSweeney KM, Bozza WP, Alterovitz W-L, Zhang B. Transcriptomic profiling reveals p53 as a key regulator of doxorubicin-induced cardiotoxicity. Cell Death Discov. 2019;5:102. https://doi.org/10.1038/s41420-019-0182-6 .
doi: 10.1038/s41420-019-0182-6
pubmed: 31231550
pmcid: 6561911
Linders AN, Dias IB, LópezFernández T, Tocchetti CG, Bomer N, Van der Meer P. A review of the pathophysiological mechanisms of doxorubicin-induced cardiotoxicity and aging. npj Aging. 2024;10:9. https://doi.org/10.1038/s41514-024-00135-7 .
doi: 10.1038/s41514-024-00135-7
pubmed: 38263284
pmcid: 10806194
Karlsson H, Fryknäs M, Larsson R, Nygren P. Loss of cancer drug activity in colon cancer HCT-116 cells during spheroid formation in a new 3-D spheroid cell culture system. Exp Cell Res. 2012;318:1577–85. https://doi.org/10.1016/j.yexcr.2012.03.026 .
doi: 10.1016/j.yexcr.2012.03.026
pubmed: 22487097
Fontoura JC, Viezzer C, dos Santos FG, Ligabue RA, Weinlich R, Puga RD, Antonow D, Severino P, Bonorino C. Comparison of 2D and 3D cell culture models for cell growth, gene expression and drug resistance. Mater Sci Eng C. 2020;107:110264. https://doi.org/10.1016/j.msec.2019.110264 .
doi: 10.1016/j.msec.2019.110264
Yip D, Cho CH. A multicellular 3D heterospheroid model of liver tumor and stromal cells in collagen gel for anti-cancer drug testing. Biochem Biophys Res Commun. 2013;433:327–32. https://doi.org/10.1016/j.bbrc.2013.03.008 .
doi: 10.1016/j.bbrc.2013.03.008
pubmed: 23501105
Azimi T, Loizidou M, Dwek MV. Cancer cells grown in 3D under fluid flow exhibit an aggressive phenotype and reduced responsiveness to the anti-cancer treatment doxorubicin. Sci Rep. 2020;10:12020. https://doi.org/10.1038/s41598-020-68999-9 .
doi: 10.1038/s41598-020-68999-9
pubmed: 32694700
pmcid: 7374750
Xia M, Huang R, Sun Y, Semenza GL, Aldred SF, Witt KL, Inglese J, Tice RR, Austin CP. Identification of chemical compounds that induce HIF-1α activity. Toxicol Sci. 2009;112:153–63. https://doi.org/10.1093/toxsci/kfp123 .
doi: 10.1093/toxsci/kfp123
pubmed: 19502547
pmcid: 2910898
Maxwell P, Salnikow K. HIF-1, an oxygen and metal responsive transcription factor. Cancer Biol Ther. 2004;3:29–35. https://doi.org/10.4161/cbt.3.1.547 .
doi: 10.4161/cbt.3.1.547
pubmed: 14726713
Wang GL, Semenza GL. General involvement of hypoxia-inducible factor 1 in transcriptional response to hypoxia. Proc Nat Acad Sci. 1993;90:4304–8. https://doi.org/10.1073/pnas.90.9.4304 .
doi: 10.1073/pnas.90.9.4304
pubmed: 8387214
pmcid: 46495
Rauen U, Springer A, Weisheit D, Petrat F, Korth H, de Groot H, Sustmann R. Assessment of chelatable mitochondrial iron by using mitochondrion-selective fluorescent iron indicators with different iron-binding affinities. ChemBioChem. 2007;8:341–52. https://doi.org/10.1002/cbic.200600311 .
doi: 10.1002/cbic.200600311
pubmed: 17219451
Matos CP, Addis Y, Nunes P, Barroso S, Alho I, Martins M, Matos APA, Marques F, Cavaco I, Costa Pessoa J, Correia I. Exploring the cytotoxic activity of new phenanthroline salicylaldimine Zn(II) complexes. J Inorg Biochem. 2019;198:110727. https://doi.org/10.1016/j.jinorgbio.2019.110727 .
doi: 10.1016/j.jinorgbio.2019.110727
pubmed: 31195153