MiR-451a and let-7i-5p loaded extracellular vesicles attenuate heme-induced inflammation in hiPSC-derived endothelial cells

Abstract

Hemolysis is associated with many pathologies, including trauma, sepsis, hemorrhagic stroke, malaria, and genetic disorders such as sickle cell disease (SCD). When hemolysis occurs, free-heme drives vascular inflammation, resulting in oxidative tissue damage and cardiometabolic complications. A better understanding of heme clearance and detoxification is essential to preventing sustained tissue damage. Human induced pluripotent stem cell (hiPSC)-derived endothelial cells (hiPSC-ECs) provide a novel source of patient-specific cells and tissues for disease modeling, drug discovery, and regenerative therapeutics. Here we report the use of hiPSC-ECs to elucidate the role of miR-451a and let-7i-5p-loaded extracellular vesicles (EVs, such as exosomes) in the inflammatory response to free-heme as a model for heme-induced inflammation. We provide evidence of a significant correlation between miR-451a and let-7i-5p-loaded circulating exosomes in plasmodium-infected patients with reported clinical benchmarks of malaria-severity (e.g., Hemoglobin (Hb) levels, white blood cell counts). Additionally, we determined that exposure of Plasmodium falciparum (Pf) parasites to EVs, loaded with either miRNA, significantly reduces their counts in vitro. Using hiPSCs derived from individuals with wild-type Hb (HbAA) or homozygous sickle cell mutated Hb (HbSS) genotypes, we demonstrate that heme-treated hiPSC-ECs secreted inflammatory products (cytokines, chemokines and growth factors) into supporting media at concentrations that were similar to that reported in HbAA and HbSS serum. This inflammatory response was attenuated by exposure with miR-451a or let-7i-5p-loaded EVs. We also found a decrease in transcription of ICAM1 and P-Selectin, as well as the secretion of key inflammatory cytokines (e.g., CXCL10, TNF-α, and IFN-γ). Based on these findings, we propose a model in which increased levels of exosomal miR-451a and let-7i-5p in Plasmodium-infected individuals will attenuate inflammatory responses to free-heme and parasite-derived products. As a result, infected erythrocytes will less likely adhere to the endothelium, sequester in brain micro vessels, and reduce vaso-occlusive crises that exacerbate cerebral malaria.

Heme-Inducing Endothelial Pyroptosis Plays a Key Role in Radiofrequency Ablation of Hepatic Hemangioma Leading to Systemic Inflammatory Response Syndrome

“…Previous studies have demonstrated that heme may induce inflammation of endothelial cells. 23 To investigate whether heme can cause SIRS by inducing endothelial cell inflammation, human lung microvascular endothelial cells (HLMVECs) were treated with heme. Heme induced morphological changes in cells after heme treatment ( Figure 3a ).…”

lnc001776 Affects CPB2 Toxin-Induced Excessive Injury of Porcine Intestinal Epithelial Cells via Activating JNK/NF-kB Pathway through ssc-let-7i-5p/IL-6 Axis

“…For instance, let-7i-5p overexpression repressed hypoxia-induced NF-κB activation, mitochondrial dysfunction and cardiomyocyte apoptosis [ 54 ]. Let-7i-5p-loaded extracellular vesicles attenuated heme-induced inflammatory injury in heme-derived endothelial cells [ 55 ]. We found that elevated ssc-let-7i-5p inhibited apoptosis, inflammatory injury and barrier dysfunction in IPEC-J2 cells caused by CPB2 toxin.…”

Modulation of Heme-Induced Inflammation Using MicroRNA-Loaded Liposomes: Implications for Hemolytic Disorders Such as Malaria and Sickle Cell Disease

“…Statistical analysis was completed using GraphPad Prism version 10.0.0 for Windows (GraphPad Software, La Jolla, CA, USA). The ∆∆CT method was used to identify mRNA gene expression [48]. Normality was checked using a Shapiro-Wilk normality test.…”

“…The final concentration of miRNA in liposomes was 400 pmol. The expression of miRNAs let-7i-5p and miR-451a was an influence on heme-induced inflammation [48].…”