Liver-Chip: Reproducing Human and Cross-Species Toxicities

Jang, K.-J.; Otieno, M.; Ronxhi, J.; Lim, H.-K.; Ewart, L.; Kodella, K.; Petropolis, D.; Kulkarni, G.; Rubins, J.; Conegliano, D.; Nawroth, J.; Simic, D.; Lam, W.; Singer, M.; Barale, E.; Singh, B.; Sonee, M.; Streeter, A.; Manthey, C.; Jones, B.; Srivastava, A.; Andersson, L.; Williams, D.; Park, H.; Barrile, R.; Sliz, J.; Herland, A.; Haney, S.; Karalis, K.; Ingber, D.; Hamilton, G.

doi:10.1101/631002

Home/Archive/bioRxiv

bioRxiv · 10.1101/631002

Liver-Chip: Reproducing Human and Cross-Species Toxicities

View original source ↗Export metadata ↓

Abstract

Nonclinical rodent and non-rodent toxicity models used to support clinical trials of candidate drugs may produce discordant results or fail to predict complications in humans contributing to drug failures in the clinic. Here we applied microengineered Organ-on-Chip (Organ-Chip) technology to design rat, dog, and human Liver-Chips containing species-specific primary hepatocytes interfaced with liver sinusoidal endothelial cells, with or without Kupffer cells and hepatic stellate cells, cultured under physiological fluid flow. The Liver-Chips detected diverse phenotypes of liver toxicity including hepatocellular injury, steatosis, cholestasis, and fibrosis as well as species-specific toxicities when treated with tool compounds. Multi-species Liver-Chips may provide a useful platform for prediction of liver toxicity and inform human relevance of liver toxicities detected in animal studies to better determine safety and human risk.\n\nOne Sentence SummaryMicroengineered Organ-Chip technology has been used to design rat, dog and human Liver-Chips that recapitulate species-specific liver toxicities.

Explore related subjects

pharmacology and toxicology Preprint

Keep this discovery

Explore connections, maps & timelines

BibTeX RIS

Jang, K.-J., Otieno, M., Ronxhi, J., Lim, H.-K., Ewart, L., Kodella, K., Petropolis, D., Kulkarni, G., Rubins, J., Conegliano, D., Nawroth, J., Simic, D., Lam, W., Singer, M., Barale, E., Singh, B., Sonee, M., Streeter, A., Manthey, C., Jones, B., Srivastava, A., Andersson, L., Williams, D., Park, H., Barrile, R., Sliz, J., Herland, A., Haney, S., Karalis, K., Ingber, D., Hamilton, G.. 2019-05-15. Liver-Chip: Reproducing Human and Cross-Species Toxicities. https://doi.org/10.1101/631002

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

bioRxiv2026

Lipid-ASO therapeutics exhibit differential tissue targeted delivery upon systemic or local CNS administration

Antisense oligonucleotides (ASOs) are a powerful therapeutic modality, but their full potential is hindered by pharmacokinetic properties that affect tissue and cellular delivery. Lipid conjugation is increasingly used to modulate ASO's biodistribution and promote extrahepatic activity, yet lipid dependent effects on in vivo functional delivery, particularly in the central nervous system (CNS), remain less explored. Here, we performed a side by side in vivo comparison of cholesterol, palmitic acid (C16:0), docosanoic acid (C22:0), and eicosapentaenoic acid (C20:5) conjugated to a fully phosphorothioated 3 10 3 LNA gapmer ASO targeting the Malat1 long non coding RNA. Lipid-ASO conjugates were administered systemically or locally in the brain of mice and evaluated for tissue level and cellular level distribution by imaging, qPCR and single-cell RNA sequencing, simultaneously annotating cell origin and global transcriptional changes within the cell. Following systemic administration in mice, lipid conjugation improved overall multi organ efficacy compared to unconjugated ASO, but with pronounced tissue specific differences. Single cell sequencing of liver and heart transcriptomes revealed lipid dependent cellular uptake patterns and transcriptional responses distinct from administration of unconjugated ASO. After intracerebroventricular administration, selected fatty acid conjugates enhanced silencing in deep brain regions such as the striatum, whereas cholesterol conjugation impaired functional delivery despite increased CNS retention. Light-sheet microscopy showed restricted parenchymal penetration of cholesterol ASOs compared with broader but heterogeneous distribution of palmitic acid conjugate. Together, these findings demonstrate that lipid identity critically determines ASO efficacy, productive cellular uptake, and regional CNS engagement, emphasizing the need for context specific lipid design in ASO therapeutic development.

pharmacology and toxicology↗

bioRxiv2026

Novel Dissymmetric Ionizable Lipid-Assembled Lipid Nanoparticles for Delivery of Ferroptosis-Related siRNA in Diabetic Treatment

Small interfering RNA (siRNA) enables precise post-transcriptional gene silencing for refractory diseases, yet its clinical translation remains limited by the lack of safe and efficient delivery vectors. Inspired by the dissymmetric alkyl chain architecture of natural membrane phospholipids, we designed and synthesized 34 novel ionizable lipids with dissymmetric hydrophobic tails and formulated them into lipid nanoparticles (LNPs). Through systematic physicochemical and biological assessments, we established clear structure-activity relationships and identified two lead LNPs (O14-LNP, H18a-LNP) with superior endosomal escape capacity, enhanced in vivo gene silencing potency, and favorable biosafety relative to the clinical benchmark MC3-LNP. In both streptozotocin-induced and spontaneous db/db type 2 diabetes (T2D) mouse models, lead LNPs delivering ferroptosis-related siRNAs effectively ameliorated glucose and lipid metabolic disorders, restored islet function, and alleviated hepatic steatosis. This study not only lays a theoretical foundation for the rational design of novel ionizable lipids, but also validates the therapeutic potential of siRNA therapy targeting ferroptosis, providing a versatile delivery platform and targeted therapeutic strategy for the treatment of T2D.

pharmacology and toxicology↗

bioRxiv2019

Levetiracetam inhibits SV2A-synaptotagmin interaction at synapses that lack SV2B

Epilepsy remains a difficult-to-treat neurological disorder prompting the need for new therapies that work via alternate mechanisms. Levetiracetam (LEV) is the first in a series of anti-epilepsy drugs that target presynaptic functioning. LEV binds the synaptic vesicle protein SV2A, and has been shown to decrease neurotransmitter release in hippocampal slices. The molecular basis of LEV action is unknown, however, and direct effects of LEV on SV2A function remain to be determined. SV2A is the most widely expressed paralog of a three-gene family (SV2A, B, C) that is variably co-expressed throughout the CNS. All three SV2s bind the calcium sensor protein synaptotagmin and SV2 plays a crucial role in synaptotagmin stability and trafficking. Here we addressed the action of LEV at the cellular and molecular level asking whether the presence of non-LEV binding SV2 paralogs influences drug action and whether LEV impacts SV2As role in synaptotagmin function. We report that LEV altered short-term synaptic plasticity in isolated neurons from SV2B knockout but not wild-type mice, mimicking the loss of SV2 function. Similarly, LEV reduced SV2A binding to synaptotagmin only in the absence of SV2B. Furthermore, LEV reduced and slowed the internalization of synaptotagmin in neurons cultured from SV2B KO but not WT mice. Taken together, these findings suggest that LEV alters synaptic release probability by disrupting SV2s regulation of synaptotagmin selectively in neurons that express only SV2A. Neurons that meet this requirement include most inhibitory neurons and the granule cells of the dentate gyrus, two classes of neuron implicated in epilepsy.

pharmacology and toxicology↗