Multi-lineage heart-chip models drug cardiotoxicity and enhances maturation of human stem cell-derived cardiovascular cells

Organ Model: Heart

Application: Toxicology

Abstract: Cardiovascular toxicity causes adverse drug reactions and may lead to drug removal from the pharmaceutical market. Cancer therapies can induce life-threatening cardiovascular side effects such as arrhythmias, muscle cell death, or vascular dysfunction. New technologies have enabled cardiotoxic compounds to be identified earlier in drug development. Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CMs) and vascular endothelial cells (ECs) can screen for drug-induced alterations in cardiovascular cell function and survival. However, most existing hiPSC models for cardiovascular drug toxicity utilize two-dimensional, immature cells grown in static culture. Improved in vitro models to mechanistically interrogate cardiotoxicity would utilize more adult-like, mature hiPSC-derived cells in an integrated system whereby toxic drugs and protective agents can flow between hiPSC-ECs that represent systemic vasculature and hiPSC-CMs that represent heart muscle (myocardium). Such models would be useful for testing the multi-lineage cardiotoxicities of chemotherapeutic drugs such as VEGFR2/PDGFR-inhibiting tyrosine kinase inhibitors (VPTKIs). Here, we develop a multi-lineage, fully-integrated, cardiovascular organ-chip that can enhance hiPSC-EC and hiPSC-CM functional and genetic maturity, model endothelial barrier permeability, and demonstrate long-term functional stability. This microfluidic organ-chip harbors hiPSC-CMs and hiPSC-ECs on separate channels that can be subjected to active fluid flow and rhythmic biomechanical stretch. We demonstrate the utility of this cardiovascular organ-chip as a predictive platform for evaluating multi-lineage VPTKI toxicity. This study may lead to the development of new modalities for the evaluation and prevention of cancer therapy-induced cardiotoxicity.

A Novel 3D Dual-Cell Perfusion Model as Tool to Identify and Validate Potential Druggable Targets to Reduce Ischemia Reperfusion Injury

Featured session at Netherlands MPS Day, which took place on 11/15/2023.

Organ-on-Chip Technology Recapitulates Thrombosis Induced by an anti-CD154 Therapeutic

Abstract

Blocking of CD40L-mediated signaling represents a validated therapeutic strategy for treatment of several auto-immune disorders, however, development of therapies against this target was stalled for several years because of unexpected thrombotic and cardiovascular events during clinical development of the anti-CD40L mAb Hu5c8. These side effects were not detected during preclinical testing. Platelet activation assays have been used to test the hypothesis that thrombosis was caused by binding of Hu5c8IgG1 to FcγRIIa receptors on platelets. To provide additional confidence in the safety of new anti-CD40L mAbs that are designed not to bind FcγRIIa, a micovessel-chip (Vessel-Chip) was developed that could capture human relevant endpoints for detection of coagulopathy, providing a patient-specific platform for safety testing. The Vessel-Chip includes a vascular channel lined by human endothelial cells and perfused with human whole blood at a physiologically-relevant shear rate. Treatment with clinical-relevant concentrations of hu5c8IgG1 and sCD40L resulted in endothelial activation, platelet adhesion, platelet aggregation, fibrin clot formation, and increased secretion of thrombin anti-thrombin (TAT) complex. Conversely, these endpoints were attenuated following treatment with Hu5c8IgG2σ, a mAb that does not bind FcγRIIa receptors. Given lack of suitable preclinical models for detection of thrombosis, these data provide confidence in the potential safety of the newer generation anti-CD40L mAbs designed not to bind FcγRIIa receptors. Detection of TAT in the model confirms that important counter-regulatory mechanisms that occur during thrombosis, such as thrombin and antithrombin generation, occur de-novo in the model. This model provides a unique platform for preclinical assessment of thrombosis risk in a patient-specific manner, but can also be used for discovery of anti-thrombotic agents, and mechanism of action elucidation thus providing a useful tool for drug discovery and development.

Monoclonal Antibody

Organ‐on‐Chip Recapitulates Thrombosis Induced by an anti‐CD154 Monoclonal Antibody: Translational Potential of Advanced Microengineered Systems

Abstract

Clinical development of Hu5c8, a monoclonal antibody against CD40L intended for treatment of autoimmune disorders, was terminated due to unexpected thrombotic complications. These life-threatening side effects were not discovered during preclinical testing due to the lack of predictive models. In the present study, we describe the development of a microengineered system lined by human endothelium perfused with human whole blood, a “Vessel-Chip.” The Vessel-Chip allowed us to evaluate key parameters in thrombosis, such as endothelial activation, platelet adhesion, platelet aggregation, fibrin clot formation, and thrombin anti-thrombin complexes in the Chip-effluent in response to Hu5c8 in the presence of soluble CD40L. Importantly, the observed prothrombotic effects were not observed with Hu5c8-IgG2σ designed with an Fc domain that does not bind the FcγRIIa receptor, suggesting that this approach may have a low potential risk for thrombosis. Our results demonstrate the translational potential of Organs-on-Chips, as advanced microengineered systems to better predict human response.