Lipopolysaccharides induced inflammatory responses and electrophysiological dysfunctions in human-induced pluripotent stem cell derived cardiomyocytes

Abstract

Severe infections like sepsis lead frequently to cardiomyopathy. The mechanisms are unclear and an optimal therapy for septic cardiomyopathy still lacks. The aim of this study is to establish an endotoxin-induced inflammatory model using human induced pluripotent stem cell (hiPSC) derived cardiomyocytes (hiPSC-CMs) for mechanistic and therapeutic studies. hiPSC-CMs were treated by lipopolysaccharide (LPS) in different concentrations for different times. ELISA, FACS, qPCR, and patch-clamp techniques were used for the study. TLR4 (Toll-like receptor 4) and its associated proteins, CD14, LBP (lipopolysaccharide binding protein), TIRAP (toll-interleukin 1 receptor domain containing adaptor protein), Ly96 (lymphocyte antigen 96) and nuclear factor kappa B as well as some pro-and anti-inflammatory factors are expressed in hiPSC-CMs. LPS-treatment for 6 hours increased the expression levels of pro-inflammatory and chemotactic cytokines (TNF-a, IL-1ß, IL-6, CCL2, CCL5, IL-8), whereas 48 hour-treatment elevated the expression of anti-inflammatory factors (IL-10 and IL-6). LPS led to cell injury resulting from exaggerated cell apoptosis and necrosis. Finally, LPS inhibited small conductance Ca2+-activated K+ channel currents, enhanced Na+/Ca2+-exchanger currents, prolonged action potential duration, suggesting cellular electrical dysfunctions. Our data demonstrate that hiPSC-CMs possess the functional reaction system involved in endotoxin-induced inflammation and can model some bacterium-induced inflammatory responses in cardiac myocytes.

Multifunctional Properties of Quercitrin-Coated Porous Ti-6Al-4V Implants for Orthopaedic Applications Assessed In Vitro

“…Another important point is to test the coating bioactivity under bacterial challenging conditions, using an LPS stimuli that increases inflammation [25], to verify if the osteoblast function is maintained. According to the anti-inflammatory properties of the flavonoids, ALP activity of osteoblasts on quercitrin-coated implants was higher than on the control, with an increase comparable to the results with the nonstimulated conditions, indicating that the osteopromotive effect was maintained even under inflammatory conditions.…”

Drug Testing in Human‐Induced Pluripotent Stem Cell–Derived Cardiomyocytes From a Patient With Short QT Syndrome Type 1

“…Our group has successfully used human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to model some key features of different diseases, including SQTS1. [22][23][24][25][26][27][28] We have demonstrated that the hiPSC-CMs from an SQTS1 patient (SQTS1 hiPSC-CMs) are able to recapitulate the single-cell phenotypic features of SQTS (APDshortening and arrhythmic calcium transients) and provide novel opportunities to further elucidate the disease mechanism and test drug effects. 27 Therefore, we designed the current study to examine the APDprolonging and antiarrhythmic effects of some drugs using our well-defined and characterized model of SQTS1.…”

“…This animal model is not ideal, either, because the differences between animals and humans—as well as the differences between the drug‐induced SQTS and disease‐induced SQTS—may influence the results of drug testing. Our group has successfully used human‐induced pluripotent stem cell–derived cardiomyocytes (hiPSC‐CMs) to model some key features of different diseases, including SQTS1 . We have demonstrated that the hiPSC‐CMs from an SQTS1 patient (SQTS1 hiPSC‐CMs) are able to recapitulate the single‐cell phenotypic features of SQTS (APD‐shortening and arrhythmic calcium transients) and provide novel opportunities to further elucidate the disease mechanism and test drug effects …”

Beyond Family: Modeling Non-hereditary Heart Diseases With Human Pluripotent Stem Cell-Derived Cardiomyocytes

“…Nevertheless, hPSC-CMs have been also employed in modeling less common pathological settings, such as systemic pathogen infections leading to myocarditis and heart failure. Modeling septic shock by exposure to bacterial lipopolysaccharides affects hPSC-CM survival, electrophysiology and demonstrates their competence in activating innate immune inflammatory responses (Yucel et al, 2017). Indeed, hPSC-CM display stronger macrophage chemo-attractant properties than purified chemokines (Pallotta et al, 2015) and significant stress-responsive paracrine proinflammatory signaling (Sebastiao et al, 2020) mediating fibrosis in vivo and in vitro (Kumar et al, 2019;Zhang et al, 2019).…”