Research ArticleDRUG TESTING

Multi-organ system for the evaluation of efficacy and off-target toxicity of anticancer therapeutics

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Science Translational Medicine  19 Jun 2019:
Vol. 11, Issue 497, eaav1386
DOI: 10.1126/scitranslmed.aav1386
  • Fig. 1 Five-chamber reconfigurable multi-organ system.

    (A) Photograph of the multi-organ system filled with green-colored dye for visualization. Scale bar, 2 cm. (B) Exploded view schematic representation of the platform assembly and design used in the system 2 study of tamoxifen. Chamber 1 houses hepatocytes on coverslips. Chambers 2 and 4 are cardiac cantilevers and microelectrode arrays (MEAs), respectively. Chambers 3 and 5 are for cancer cells SW-962 and MCF-7. Drugs were applied to medium access port A and initially passed over the liver to mimic aspects of first pass metabolism.

  • Fig. 2 Kasumi-1 and megakaryocyte (MEG-01) proliferation over 14 days.

    (A) Representative phase contrast images of Kasumi-1 and megakaryocytes growing inside the multi-organ system chamber on days in vitro (DIV) 1, 7, and 14. (B) Quantification of proliferation, represented as both a linear fold change and a log-transformed change in cell number over time. Error bars are represented as SD of at least six replicates across a minimum of three biological replicates.

  • Fig. 3 Effect of imatinib and diclofenac on proliferative capability of megakaryocyte and Kasumi-1 cells.

    (A) MEG-01 and (B) Kasumi-1 cells in bone marrow/liver system under continuous flow. Drug addition was performed on day 4, and effects on proliferation were evaluated at DIV 7 (day 3 after drug addition). Error bars are represented as SEM. *P < 0.02 [one-way analysis of variance (ANOVA) followed by Dunnett’s test (α = 0.05)] and represents a minimum of n = 4 biological replicates.

  • Fig. 4 Liver functionality and viability in bone marrow/liver system.

    End point measurements of CYP isoforms 3A4 (A), 1A1 (B), and 2C9 (C). (D) Relative viability after 3 days of drug exposure. Liver viability was assessed after drug exposure to determine drug effects on liver function and survival at DIV 7. Error bars are represented as SEM. CYP values are double-normalized to control values and to cell number. *P < 0.003 and **P < 0.0002 [one-way ANOVA followed by Dunnett’s test (α = 0.05)] and represent a minimum of n = 3 biological replicates.

  • Fig. 5 Tamoxifen treatment effects on cancer cell viability in the presence and absence of liver tissue.

    (A) Normalized viability of MCF-7 cells treated with 4-hydroxytamoxifen or tamoxifen with or without verapamil in the presence or absence of liver tissue. (B) Normalized viability of SW-962 cancer cells, which express Pgp, treated with 4-hydroxytamoxifen or tamoxifen with or without verapamil in the presence or absence of liver tissue. Error bars are represented as SEM. *P < 0.02 [one-way ANOVA followed by Tukey’s test (α = 0.05)] and represents a minimum of n = 4 biological replicates.

  • Fig. 6 Effects of tamoxifen on cardiac function.

    Normalized (A) CV, (B) cardiac force, (C) beat frequency, and (D) cell viability of cardiac cells treated with tamoxifen with or without verapamil. Error bars are represented as SEM. *P < 0.02 [one-way (A) or two-way (B, C, and D) ANOVA followed by Tukey’s test (α = 0.05)] and represents a minimum of n = 3 biological replicates.

  • Fig. 7 CFD modeling and HPLC drug analysis in a multi-organ BoaC system.

    (A) CFD modeling was performed to determine PD parameters, incorporating cyclic rocking flow, drug metabolism by liver cells, and complexing of drug and albumin. (B) A time-dependent HPLC analysis was performed on sample aliquots drawn from the system to generate an AUC tamoxifen concentration profile within the system 2 configuration.

Supplementary Materials

  • stm.sciencemag.org/cgi/content/full/11/497/eaav1386/DC1

    Fig. S1. Overview of four-organ system with cancer, cardiac, and liver and phase contrast microscopy of tissues at 14 days.

    Fig. S2. Escalating dose of verapamil effect on MCF-7 and SW-962 viability.

    Fig. S3. In situ functional cardiac and liver readouts after 14 days.

    Table S1. Formulation of human iPSC-derived cardiomyocyte culture medium.

    Table S2. Formulation of primary human liver culture medium.

    Table S3. Formulation of multi-organ system common medium.

    Data file S1. Primary data.

  • The PDF file includes:

    • Fig. S1. Overview of four-organ system with cancer, cardiac, and liver and phase contrast microscopy of tissues at 14 days.
    • Fig. S2. Escalating dose of verapamil effect on MCF-7 and SW-962 viability.
    • Fig. S3. In situ functional cardiac and liver readouts after 14 days.
    • Table S1. Formulation of human iPSC-derived cardiomyocyte culture medium.
    • Table S2. Formulation of primary human liver culture medium.
    • Table S3. Formulation of multi-organ system common medium.

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    Other Supplementary Material for this manuscript includes the following:

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