Kim-1 Targeted Extracellular Vesicles: A New Therapeutic Platform for RNAi to Treat AKI

Abstract

Background: Acute kidney injury (AKI) is a significant public health problem with high morbidity and mortality. Unfortunately, no definitive treatment is currently available for AKI. RNA interference (RNAi) provides a new and potent method for gene therapy to tackle this dilemma. Methods We engineered red blood cell-derived extracellular vesicles (REVs) with targeting peptides and therapeutic siRNAs to treat experimental AKI in a mouse model following renal ischemia/reperfusion (I/R) injury and unilateral ureteral obstruction (UUO). Phage display identified peptides that bind to the kidney injury molecule-1 (Kim-1). RNA-sequencing (RNA-seq) characterized the transcriptome of ischemic kidney to explore potential therapeutic targets. Results REVs targeted with Kim-1-binding LTH peptide (REVLTH) efficiently homed to and accumulated at the injured tubules in kidney following I/R injury. We identified transcription factors P65 and Snai1 that drive inflammation and fibrosis as potential therapeutic targets. Taking advantage of the established REVLTH, siRNAs targeting P65 and Snai1 were efficiently delivered to ischemic kidney and consequently blocked the expression of P-p65 and Snai1 in tubules. Moreover, dual suppression of P65 and Snai1 significantly improved I/R- and UUO-induced kidney injury by alleviating tubulointerstitial inflammation and fibrosis, and potently abrogated the transition to chronic kidney disease. Conclusions A red blood cell-derived extracellular vesicle platform targeted Kim-1 in acutely injured mouse kidney and delivered siRNAs for transcription factors P65 and Snai1, alleviating inflammation and fibrosis in the tubules.

Lipid-Based Nanovesicular Drug Delivery Systems

“…Suppress postoperative breast cancer metastasis [370] EVs from murine neuroblastoma cell line and dendritic cells Cholesterol-conjugated siRNAs Human antigen R silencing for cancer treatment [334] Exosomes from HEK-293T cells c-Met siRNA Reverse chemoresistance to cisplatin in gastric cancer [371] Exosomes from HEK-293T cells Hepatocyte growth factor (HGF) siRNA Inhibitory effect on tumor growth and angiogenesis in gastric cancer [372] EVs from mesenchymal stem cells derived from umbilical cord Wharton's jelly Hydrophobically modified asymmetric siRNAs conjugated with cholesterol Huntingtin silencing in neurons [333] Exosomes from glioblastoma cells [373] Exosomes from human neuroblastoma cells Heat shock protein-27 (HSP27) siRNA Decrease of cell differentiation toward mature neuron in neuroblastoma [374] Exosomes from urine-derived induced pluripotent stem cells ICAM-1 siRNA Alleviating inflammation of pulmonary microvascular endothelial cells [375] Exosomes from HEK-293T cells KRAS siRNA Inhibition of tumor growth [376] EVs from astrocytes LincRNA-Cox2 siRNA Lipopolysaccharideinduced microglial proliferation for treatment of CNS disorders [377] Exosomes from mesenchymal stem cells PTEN siRNA Promote recovery for spinal cord injury individuals [378] EVs from red blood cells P65 and Snai1 siRNA Inhibit renal inflammation and fibrosis for acute kidney injury treatment [379] EVs from HEK-293T cells RAGE siRNA Attenuated inflammation in myocarditis [380] Exosomes from bone-marrow-derived mesenchymal stem cells siGRP78 Suppress Sorafenib resistance in hepatocellular carcinoma [381] Exosomes from bovine milk siKRAS Lung tumor treatment [382] EVs from different cell lines siRNA Reducing the therapeutic dose of siRNA for different pathologies [383] EVs from human umbilical cord mesenchymal stem cells siRNA-ELFN1-AS1 Inhibit colon adenocarcinoma cells proliferation [384] Exosomes from normal human foreskin fibroblast siRNA or short hairpin RNA specific to oncogenic Kras G12D Pancreatic ductal adenocarcinoma treatment [385] Exosomes from HEK-293T cells Transient receptor potential polycystic 2 (TRPP2) siRNA…”

Extracellular Vesicles in Acute Kidney Injury and Clinical Applications

“…These include a fusion of the exosome membrane protein LAMP2B to the neuron-specific RVG peptide 3 to improve neuronal delivery [133], an enhanced stable retention of exosomes in circulation by expressing CD47 in MSCs [134], and the development of a novel molecular platform called ARMS (Arrestin domain containing protein 1-mediated microvesicles) to improve cargo packaging and delivery [135]. In this vein, a recent development of the specific delivery of erythrocyte-derived EVs charged with therapeutic small interfering RNAs (siRNAs) to the damaged kidney with AKI is remarkable [136]. The key to this feat was the development of synthetic peptides that bound to the AKI biomarker KIM-1 [136].…”

“…In this vein, a recent development of the specific delivery of erythrocyte-derived EVs charged with therapeutic small interfering RNAs (siRNAs) to the damaged kidney with AKI is remarkable [136]. The key to this feat was the development of synthetic peptides that bound to the AKI biomarker KIM-1 [136]. Expressing the peptides on the EVs derived from erythrocytes delivered the EVs charged with therapeutic siRNAs specifically to the AKI kidney, ameliorating tubulointerstitial inflammation and fibrosis [136].…”

“…The key to this feat was the development of synthetic peptides that bound to the AKI biomarker KIM-1 [136]. Expressing the peptides on the EVs derived from erythrocytes delivered the EVs charged with therapeutic siRNAs specifically to the AKI kidney, ameliorating tubulointerstitial inflammation and fibrosis [136].…”

Riclinoctaose Attenuates Renal Ischemia-Reperfusion Injury by the Regulation of Macrophage Polarization

“…Ischemic preconditioning consists of cycles of mild ischemic stress, which elicit tolerance against subsequent severe ischemia and is an essential tool for clinical management of IRI (Livingston et al, 2019). Gene therapy, such as RNA interference targeting transcription factors that drive inflammation and fibrosis, has also been reported to be therapeutically effective in experimental renal IRI (Haddad et al, 2021;Tang et al, 2021;Zhuang et al, 2021). The therapeutic efficacy of the RNA interference is dependent on its stability, which increases the difficulty and cost of clinical utility.…”