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Current status and future prospects of toxicity assessment using organoids

  • Received : 2024.11.10
  • Accepted : 2025.05.18
  • Published : 2025.07.15

Abstract

Organoids offer innovative platforms for toxicity assessment by more accurately recapitulating the structural and functional complexity of human tissues compared to traditional 2D cultures and animal models. This review summarizes current applications and discusses future directions for kidney, cardiac, liver, and brain organoids within the context of toxicology. Brain organoids, which recapitulate key features of human neural development, have facilitated investigations into neurotoxicity induced by agents such as Zika virus and chlorpyrifos. Kidney organoids, with nephron-like structures, have been employed to model nephrotoxicity induced by chemotherapeutic agents. Cardiac organoids, which mimic heart contractions and electrical properties, are effective for assessing cardiotoxicity from drugs such as doxorubicin. Liver organoids allow prediction of hepatotoxicity through the modeling of human-specific drug metabolism, as shown in studies using compounds like acetaminophen and troglitazone. Despite their promise, organoid systems still face challenges such as cellular immaturity, batch-to-batch variability, and limited vascularization. Emerging technologies such as 3D bioprinting, vascular integration, and multi-organ assembly are expected to improve the applicability of organoids in toxicity testing. Additionally, integration with microfluidic platforms and artificial intelligence-based analysis will improve high-throughput screening and predictive accuracy. As these technologies continue to evolve, organoids are poised to play a critical role in safer drug development, reducing dependence on animal models and providing deeper insights into systemic toxicity.

Keywords

Acknowledgement

This work was supported by the Technology Innovation Program (or Industrial Strategic Technology Development Program-3D-TissueChip Based Drug Discovery Platform Program) (20009773, Commercialization of 3D Multifunction Tissue Mimetics Based Drug Evaluation Platform) funded by the Ministry of Trade, Industry & Energy (MOTIE, Korea).

References

  1. Kapałczyńska M, Kolenda T, Przybyła W, Zajączkowska M, Teresiak A, Filas V, Ibbs M, Bliźniak R, Łuczewski Ł, Lamperska K (2018) 2D and 3D cell cultures - a comparison of different types of cancer cell cultures. Arch Med Sci 14:910–919. https://doi.org/10.5114/aoms.2016.63743
  2. Griffith LG, Swartz MA (2006) Capturing complex 3D tissue physiology in vitro. Nat Rev Mol Cell Biol 7:211–224. https://doi.org/10.1038/nrm1858
  3. Yamada KM, Cukierman E (2007) Modeling tissue morphogenesis and cancer in 3d. Cell 130:601–610. https://doi.org/10.1016/j.cell.2007.08.006
  4. Breslin S, O'Driscoll L (2013) Three-dimensional cell culture: the missing link in drug discovery. Drug Discov Today 18:240–249. https://doi.org/10.1016/j.drudis.2012.10.003
  5. Jensen C, Teng Y (2020) Is it time to start transitioning from 2D to 3D cell culture? Front Mol Biosci 7:33. https://doi.org/10.3389/fmolb.2020.00033
  6. Brooks A, Liang X, Zhang Y, Zhao CX, Roberts MS, Wang H, Zhang L, Crawford DHG (2021) Liver organoid as a 3D in vitro model for drug validation and toxicity assessment. Pharmacol Res 169:105608. https://doi.org/10.1016/j.phrs.2021.105608
  7. Juarez-Moreno K, Chávez-García D, Hirata G, Vazquez-Duhalt R (2022) Monolayer (2D) or spheroids (3D) cell cultures for nanotoxicological studies? Comparison of cytotoxicity and cell internalization of nanoparticles. Toxicol In Vitro 85:105461. https://doi.org/10.1016/j.tiv.2022.105461
  8. Lancaster MA, Knoblich JA (2014) Generation of cerebral organoids from human pluripotent stem cells. Nat Protoc 9:2329–2340. https://doi.org/10.1038/nprot.2014.158
  9. Shi Y, Sun L, Wang M, Liu J, Zhong S, Li R, Li P, Guo L, Fang A, Chen R, Ge WP, Wu Q, Wang X (2020) Vascularized human cortical organoids (vOrganoids) model cortical development in vivo. PLoS Biol 18:e3000705. https://doi.org/10.1371/journal.pbio.3000705
  10. Nickels SL, Modamio J, Mendes-Pinheiro B, Monzel AS, Betsou F, Schwamborn JC (2020) Reproducible generation of human midbrain organoids for in vitro modeling of Parkinson's disease. Stem Cell Res 46:101870. https://doi.org/10.1016/j.scr.2020.101870
  11. Huang WK, Wong SZH, Pather SR, Nguyen PTT, Zhang F, Zhang DY, Zhang Z, Lu L, Fang W, Chen L, Fernandes A, Su Y, Song H, Ming GL (2021) Generation of hypothalamic arcuate organoids from human induced pluripotent stem cells. Cell Stem Cell 28:1657–1670.e10. https://doi.org/10.1016/j.stem.2021.04.006
  12. Cakir B, Xiang Y, Tanaka Y, Kural MH, Parent M, Kang YJ, Chapeton K, Patterson B, Yuan Y, He CS, Raredon MSB, Dengelegi J, Kim KY, Sun P, Zhong M, Lee S, Patra P, Hyder F, Niklason LE, Lee SH, Yoon YS, Park IH (2019) Engineering of human brain organoids with a functional vascular-like system. Nat Methods 16:1169–1175. https://doi.org/10.1038/s41592-019-0586-5
  13. Mansour AA, Goncalves JT, Bloyd CW, Li H, Fernandes S, Quang D, Johnston S, Parylak SL, Jin X, Gage FH (2018) An in vivo model of functional and vascularized human brain organoids. Nat Biotechnol 36:432–441. https://doi.org/10.1038/nbt.4127
  14. Chu JTS, Lamers MM (2024) Organoids in virology. NPJ Viruses 2:5. https://doi.org/10.1038/s44298-024-00017-5
  15. Sasaki M, Uemura K, Sato A, Toba S, Sanaki T, Maenaka K, Hall WW, Orba Y, Sawa H (2021) SARS-CoV-2 variants with mutations at the S1/S2 cleavage site are generated in vitro during propagation in TMPRSS2-deficient cells. PLoS Pathog 17:e1009233. https://doi.org/10.1371/journal.ppat.1009233
  16. Garcez PP, Loiola EC, Madeiro da Costa R, Higa LM, Trindade P, Delvecchio R, Nascimento JM, Brindeiro R, Tanuri A, Rehen SK (2016) Zika virus impairs growth in human neurospheres and brain organoids. Science 352:816–818. https://doi.org/10.1126/science.aaf6116
  17. Wang Y, Wang L, Zhu Y, Qin J (2018) Human brain organoid-on-a-chip to model prenatal nicotine exposure. Lab Chip 18:851–860. https://doi.org/10.1039/c7lc01084b
  18. Yin F, Zhu Y, Wang Y, Qin J (2018) Engineering brain organoids to probe impaired neurogenesis induced by cadmium. ACS Biomater Sci Eng 4:1908–1915. https://doi.org/10.1021/acsbiomaterials.8b00160
  19. Modafferi S, Zhong X, Kleensang A, Murata Y, Fagiani F, Pamies D, Hogberg HT, Calabrese V, Lachman H, Hartung T, Smirnova L (2021) Gene-environment interactions in developmental neurotoxicity: a case study of synergy between Chlorpyrifos and CHD8 knockout in human brainspheres. Environ Health Perspect 129:77001. https://doi.org/10.1289/EHP8580
  20. Cook D, Brown D, Alexander R, March R, Morgan P, Satterthwaite G, Pangalos MN (2014) Lessons learned from the fate of AstraZeneca's drug pipeline: a five-dimensional framework. Nat Rev Drug Discov 13:419–431. https://doi.org/10.1038/nrd4309
  21. Liu F, Huang J, Liu Z (2019) Vincristine impairs microtubules and causes neurotoxicity in cerebral organoids. Neuroscience 404:530–540. https://doi.org/10.1016/j.neuroscience.2018.12.047
  22. Cui K, Wang Y, Zhu Y, Tao T, Yin F, Guo Y, Liu H, Li F, Wang P, Chen Y, Qin J (2020) Neurodevelopmental impairment induced by prenatal valproic acid exposure shown with the human cortical organoid-on-a-chip model. Microsyst Nanoeng 6:49. https://doi.org/10.1038/s41378-020-0165-z
  23. Pham MT, Pollock KM, Rose MD, Cary WA, Stewart HR, Zhou P, Nolta JA, Waldau B (2018) Generation of human vascularized brain organoids. NeuroReport 29:588–593. https://doi.org/10.1097/WNR.0000000000001014
  24. Shi H, Kowalczewski A, Vu D, Liu X, Salekin A, Yang H, Ma Z (2024) Organoid intelligence: integration of organoid technology and artificial intelligence in the new era of in vitro models. Med Nov Technol Devices. https://doi.org/10.1016/j.medntd.2023.100276
  25. Alam El Din DM, Shin J, Lysinger A, Roos MJ, Johnson EC, Shafer TJ, Hartung T, Smirnova L (2024) Organoid intelligence for developmental neurotoxicity testing. Front Cell Neurosci 18:1480845. https://doi.org/10.3389/fncel.2024.1480845
  26. Kim JI, Imaizumi K, Thete MV, Hudacova Z, Jurjuţ O, Amin ND, Scherrer G, Paşca SP (2025). Human assembloid model of the ascending neural sensory pathway. Nature. https://doi.org/10.1038/s41586-025-08808-3
  27. Takasato M, Er PX, Becroft M, Vanslambrouck JM, Stanley EG, Elefanty AG, Little MH (2014) Directing human embryonic stem cell differentiation towards a renal lineage generates a self-organizing kidney. Nat Cell Biol 16:118–126. https://doi.org/10.1038/ncb2894
  28. Taguchi A, Nishinakamura R (2017) Higher-order kidney organogenesis from pluripotent stem cells. Cell Stem Cell 21:730–746.e6. https://doi.org/10.1016/j.stem.2017.10.011
  29. Homan KA, Kolesky DB, Skylar-Scott MA, Herrmann J, Obuobi H, Moisan A, Lewis JA (2016) Bioprinting of 3D convoluted renal proximal tubules on perfusable chips. Sci Rep 6:34845. https://doi.org/10.1038/srep34845
  30. Freedman BS, Brooks CR, Lam AQ, Fu H, Morizane R, Agrawal V, Saad AF, Li MK, Hughes MR, Werff RV, Peters DT, Lu J, Baccei A, Siedlecki AM, Valerius MT, Musunuru K, McNagny KM, Steinman TI, Zhou J, Lerou PH, Bonventre JV (2015) Modelling kidney disease with CRISPR-mutant kidney organoids derived from human pluripotent epiblast spheroids. Nat Commun 6:8715. https://doi.org/10.1038/ncomms9715
  31. Yousef Yengej FA, Jansen J, Rookmaaker MB, Verhaar MC, Clevers H (2020) Kidney organoids and tubuloids. Cells. https://doi.org/10.3390/cells9061326
  32. Hirayama R, Toyohara K, Watanabe K, Otsuki T, Araoka T, Mae SI, Horinouchi T, Yamamura T, Okita K, Hotta A, Iijima K, Nozu K, Osafune K (2023) iPSC-derived type IV collagen α5-expressing kidney organoids model Alport syndrome. Commun Biol 6:854. https://doi.org/10.1038/s42003-023-05203-4
  33. Cui S, Fang X, Lee H, Shin YJ, Koh ES, Chung S, Park HS, Lim SW, Lee KI, Lee JY, Yang CW, Chung BH (2023) Modeling of Fabry disease nephropathy using patient-derived human induced pluripotent stem cells and kidney organoid system. J Transl Med 21:138. https://doi.org/10.1186/s12967-023-03992-0
  34. Zarate YA, Hopkin RJ (2008) Fabry's disease. Lancet 372:1427–1435. https://doi.org/10.1016/S0140-6736(08)61589-5
  35. Vormann MK, Vriend J, Lanz HL, Gijzen L, van den Heuvel A, Hutter S, Joore J, Trietsch SJ, Stuut C, Nieskens TTG, Peters JGP, Ramp D, Caj M, Russel FGM, Jacobsen B, Roth A, Lu S, Polli JW, Naidoo AA, Vulto P, Masereeuw R, Wilmer MJ, Suter-Dick L (2021) Implementation of a human renal proximal tubule-on-a-chip for nephrotoxicity and drug interaction studies. J Pharm Sci 110:1601–1614. https://doi.org/10.1016/j.xphs.2021.01.028
  36. Thevenod F, Friedmann JM (1999) Cadmium-mediated oxidative stress in kidney proximal tubule cells induces degradation of Na+/K(+)-ATPase through proteasomal and endo-/lysosomal proteolytic pathways. FASEB J 13:1751–1761. https://doi.org/10.1096/fasebj.13.13.1751
  37. Vidal Yucha SE, Quackenbush D, Chu T, Lo F, Sutherland JJ, Kuzu G, Roberts C, Luna F, Barnes SW, Walker J, Kuss P (2022) 3D, human renal proximal tubule (RPTEC-TERT1) organoids 'tubuloids' for translatable evaluation of nephrotoxins in high-throughput. PLoS ONE 17:e0277937. https://doi.org/10.1371/journal.pone.0277937
  38. Gv M, Faria J, Sendino Garvi E, Janssen MJ, Masereeuw R, Mihaila SM (2022) Organs-on-chip technology: a tool to tackle genetic kidney diseases. Pediatr Nephrol 37:2985–2996. https://doi.org/10.1007/s00467-022-05508-2
  39. Yang Z, Zhang Y, Wang J, Yin J, Wang Z, Pei R (2023) Cardiac organoid: multiple construction approaches and potential applications. J Mater Chem B 11:7567–7581. https://doi.org/10.1039/d3tb00783a
  40. Ni B, Ye L, Zhang Y, Hu S, Lei W (2025) Advances in humanoid organoid-based research on inter-organ communications during cardiac organogenesis and cardiovascular diseases. J Transl Med 23:380. https://doi.org/10.1186/s12967-025-06381-x
  41. Lian X, Hsiao C, Wilson G, Zhu K, Hazeltine LB, Azarin SM, Raval KK, Zhang J, Kamp TJ, Palecek SP (2012) Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling. Proc Natl Acad Sci U S A 109:E1848–E1857. https://doi.org/10.1073/pnas.1200250109
  42. Breckwoldt K, Letuffe-Breniere D, Mannhardt I, Schulze T, Ulmer B, Werner T, Benzin A, Klampe B, Reinsch MC, Laufer S, Shibamiya A, Prondzynski M, Mearini G, Schade D, Fuchs S, Neuber C, Kramer E, Saleem U, Schulze ML, Rodriguez ML, Eschenhagen T, Hansen A (2017) Differentiation of cardiomyocytes and generation of human engineered heart tissue. Nat Protoc 12:1177–1197. https://doi.org/10.1038/nprot.2017.033
  43. Chen X, Lu N, Huang S, Zhang Y, Liu Z, Wang X (2023) Assessment of doxorubicin toxicity using human cardiac organoids: a novel model for evaluating drug cardiotoxicity. Chem Biol Interact 386:110777. https://doi.org/10.1016/j.cbi.2023.110777
  44. Burridge PW, Matsa E, Shukla P, Lin ZC, Churko JM, Ebert AD, Lan F, Diecke S, Huber B, Mordwinkin NM, Plews JR, Abilez OJ, Cui B, Gold JD, Wu JC (2014) Chemically defined generation of human cardiomyocytes. Nat Methods 11:855–860. https://doi.org/10.1038/nmeth.2999
  45. Hulot JS (2019) Modeling cardiac arrhythmias with organoids. J Am Coll Cardiol 73:2325–2327. https://doi.org/10.1016/j.jacc.2019.01.076
  46. Hoang P, Kowalczewski A, Sun S, Winston TS, Archilla AM, Lemus SM, Ercan-Sencicek AG, Gupta AR, Liu W, Kontaridis MI, Amack JD, Ma Z (2021) Engineering spatial-organized cardiac organoids for developmental toxicity testing. Stem Cell Reports 16:1228–1244. https://doi.org/10.1016/j.stemcr.2021.03.013
  47. Teles D, Kim Y, Ronaldson-Bouchard K, Vunjak-Novakovic G (2021) Machine learning techniques to classify healthy and diseased cardiomyocytes by contractility profile. ACS Biomater Sci Eng 7:3043–3052. https://doi.org/10.1021/acsbiomaterials.1c00418
  48. Skardal A, Murphy SV, Devarasetty M, Mead I, Kang HW, Seol YJ, Zhang YS, Shin SR, Zhao L, Aleman J, Hall AR, Shupe TD, Kleensang A, Dokmeci MR, Lee SJ, Jackson JD, Yoo JJ, Hartung T, Khademhosseini A, Soker S, Bishop CE, Atala A (2017) Multi-tissue interactions in an integrated three-tissue organ-on-a-chip platform. Sci Rep 7:8837. https://doi.org/10.1038/s41598-017-08879-x
  49. Takebe T, Sekine K, Enomura M, Koike H, Kimura M, Ogaeri T, Zhang RR, Ueno Y, Zheng YW, Koike N, Aoyama S, Adachi Y, Taniguchi H (2013) Vascularized and functional human liver from an iPSC-derived organ bud transplant. Nature 499:481–484. https://doi.org/10.1038/nature12271
  50. Sun XC, Kong DF, Zhao J, Faber KN, Xia Q, He K (2023) Liver organoids: established tools for disease modeling and drug development. Hepatol Commun. https://doi.org/10.1097/HC9.0000000000000105
  51. Shinozawa T, Kimura M, Cai Y, Saiki N, Yoneyama Y, Ouchi R, Koike H, Maezawa M, Zhang RR, Dunn A, Ferguson A, Togo S, Lewis K, Thompson WL, Asai A, Takebe T (2021) High-fidelity drug-induced liver injury screen using human pluripotent stem cell-derived organoids. Gastroenterology 160:831–846.e10. https://doi.org/10.1053/j.gastro.2020.10.002
  52. Zhang C, Zhang Q, Li J, Yu L, Li F, Li W, Li Y, Peng H, Zhao J, Carmichael PL, Wang Y, Peng S, Guo J (2020) Integration of in vitro data from three dimensionally cultured HepaRG cells and physiologically based pharmacokinetic modeling for assessment of acetaminophen hepatotoxicity. Regul Toxicol Pharmacol 114:104661. https://doi.org/10.1016/j.yrtph.2020.104661
  53. Ramli MNB, Lim YS, Koe CT, Demircioglu D, Tng W, Gonzales KAU, Tan CP, Szczerbinska I, Liang H, Soe EL, Lu Z, Ariyachet C, Yu KM, Koh SH, Yaw LP, Jumat NHB, Lim JSY, Wright G, Shabbir A, Dan YY, Ng HH, Chan YS (2020) Human pluripotent stem cell-derived organoids as models of liver disease. Gastroenterology 159:1471–1486.e12. https://doi.org/10.1053/j.gastro.2020.06.010
  54. Cheng W, Zhou Y, Xie Y, Li Y, Zhou R, Wang H, Feng Y, Wang Y (2023) Combined effect of polystyrene microplastics and bisphenol A on the human embryonic stem cells-derived liver organoids: the hepatotoxicity and lipid accumulation. Sci Total Environ 854:158585. https://doi.org/10.1016/j.scitotenv.2022.158585
  55. Li Y, Nie Y, Yang X, Liu Y, Deng X, Hayashi Y, Plummer R, Li Q, Luo N, Kasai T, Okumura T, Kamishibahara Y, Komoto T, Ohkuma T, Okamoto S, Isobe Y, Yamaguchi K, Furukawa Y, Taniguchi H (2024) Integration of Kupffer cells into human iPSC-derived liver organoids for modeling liver dysfunction in sepsis. Cell Rep 43:113918. https://doi.org/10.1016/j.celrep.2024.113918
  56. Nguyen VVT, Ye S, Gkouzioti V, van Wolferen ME, Yengej FY, Melkert D, Siti S, de Jong B, Besseling PJ, Spee B, van der Laan LJW, Horland R, Verhaar MC, van Balkom BWM (2022) A human kidney and liver organoid-based multi-organ-on-a-chip model to study the therapeutic effects and biodistribution of mesenchymal stromal cell-derived extracellular vesicles. J Extracell Vesicles 11:e12280. https://doi.org/10.1002/jev2.12280