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An Expandable Mechanopharmaceutical Device (2): Drug Induced Granulomas Maximize the Cargo Sequestering Capacity of Macrophages in the Liver

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Abstract

Purpose

Drug-induced liver injuries (DILI) comprise a significant proportion of adverse drug reactions leading to hospitalizations and death. One frequent DILI is granulomatous inflammation from exposure to harmful metabolites that activate inflammatory pathways of immune cells of the liver, which may act as a barrier to isolate the irritating stimulus and limit tissue damage.

Methods

Paralleling the accumulation of CFZ precipitates in the liver, granulomatous inflammation was studied to gain insight into its effect on liver structure and function. A structural analog that does not precipitate within macrophages was also studied using micro-analytical approaches. Depleting macrophages was used to inhibit granuloma formation and assess its effect on drug bioaccumulation and toxicity.

Results

Granuloma-associated macrophages showed a distinct phenotype, differentiating them from non-granuloma macrophages. Granulomas were induced by insoluble CFZ cargo, but not by the more soluble analog, pointing to precipitation being a factor driving granulomatous inflammation. Granuloma-associated macrophages showed increased activation of lysosomal master-regulator transcription factor EB (TFEB). Inhibiting granuloma formation increased hepatic necrosis and systemic toxicity in CFZ-treated animals.

Conclusions

Granuloma-associated macrophages are a specialized cell population equipped to actively sequester and stabilize cytotoxic chemotherapeutic agents. Thus, drug-induced granulomas may function as drug sequestering “organoids” –an induced, specialized sub-compartment– to limit tissue damage.

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Abbreviations

ADR:

Adverse Drug Reaction

CFZ:

Clofazimine

CLDI:

Crystal-Like Drug Inclusion

DILI:

Drug-induced liver injury

IL-1RA:

Interleukin-1 Receptor Antagonist

LAMP1:

Lysosomal Associated Membrane Protein 1

LC3:

Microtubule-associated protein 1A/1B-light chain 3

TFEB:

Transcription Factor EB

TLR9:

Toll-Like Receptor 9

TUNEL:

Terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling

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Acknowledgments and Disclosures

The authors thank Eric Bushong and Mark Ellisman for helping us with electron microscopy analysis done at the National Center for Microscopy and Imaging Research, University of California San Diego. The authors would also like to acknowledge support received from the Upjohn Research Award presented to GRR by the University Of Michigan College Of Pharmacy. This work was supported by NIH grant R01GM078200 to GRR.

Author information

Authors and Affiliations

  1. Department of Pharmaceutical Sciences, College of Pharmacy, University of Michigan, 428 Church Street, Ann Arbor, Michigan, 48109, USA

    Phillip Rzeczycki, Gi Sang Yoon, Rahul K. Keswani, Sudha Sud, Jason Baik, Mikhail D. Murashov & Gus R. Rosania

  2. Unit for Laboratory Animal Medicine, Medical School Office of Research, University of Michigan, 2800 Plymouth Road, Ann Arbor, Michigan, 48109, USA

    Ingrid L. Bergin

  3. Department of Clinical Pharmacy, College of Pharmacy, University of Michigan, 428 Church Street, Ann Arbor, Michigan, 48104, USA

    Kathleen A. Stringer

  4. University of Michigan College of Pharmacy, Ann Arbor, Michigan, 48109, USA

    Gus R. Rosania

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  1. Phillip Rzeczycki
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Contributions

P.R., G.S.Y, J.B., K.A.S. and G.R.R designed the research. P.R., G.S.Y., J.B., E.B., and M.E. conducted the experiments. G.R.R. and K.A.S. contributed new reagents and analytical tools. P.R., G.S.Y and I. B. analyzed the data. P.R., G.S.Y., I.B., K.A.S. and G.R.R. wrote or contributed to the writing of the manuscript.

Corresponding author

Correspondence to Gus R. Rosania.

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Rzeczycki, P., Yoon, G.S., Keswani, R.K. et al. An Expandable Mechanopharmaceutical Device (2): Drug Induced Granulomas Maximize the Cargo Sequestering Capacity of Macrophages in the Liver. Pharm Res 36, 3 (2019). https://doi.org/10.1007/s11095-018-2541-z

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