Cyclo-RGD Truncated Polymeric Nanoconstruct with Dendrimeric Templates for Targeted HDAC4 Gene Silencing in a Diabetic Nephropathy Mouse Model

Abstract

Diabetic nephropathy (DN), a chronic progressive kidney disease, is a significant complication of diabetes mellitus. Dysregulation of the histone deacetylases (HDACs) gene has been implicated in the pathogenesis of DN. Hence, the HDAC-inhibitors have emerged as a critical class of therapeutic agents in DN; however, the currently available HDAC4-inhibitors are mostly nonselective in nature as well as inhibit multiple HDACs. RNA interference of HDAC4 (HDAC4 siRNA) has shown immense promise, but the clinical translation has been impeded due to lack of a targeted, specific, and in vivo applicable delivery modality. In the present investigation, we examined Cyclo(RGDfC) (cRGD) truncated polymeric nanoplex with dendrimeric templates for targeted HDAC4 Gene Silencing. The developed nanoplex exhibited enhanced encapsulation of siRNA and offered superior protection against serum RNase nucleases degradation. The nanoplex was tested on podocytes (in vitro), wherein it showed selective binding to the αvβ3 integrin receptor, active cellular uptake, and significant in vitro gene silencing. The in vivo experiments showed remarkable suppression of the HDAC4 and inhibition in the progression of renal fibrosis in the Streptozotocin (STZ) induced DN C57BL/6 mice model. Histopathological and toxicological studies revealed nonsignificant abnormality/toxicity with the nanoplex. Conclusively, nanoplex was found as a promising tactic for targeted therapy of podocytes and could be extended for other kidney-related ailments.

Inhibition of HDAC6 With CAY10603 Ameliorates Diabetic Kidney Disease by Suppressing NLRP3 Inflammasome

“…HDAC4 contributes to podocyte injury and links renal injury to autophagy in DN, and in vivo gene silencing of HDAC4 reduces podocyte injury and proteinuria in diabetic rats ( Wang et al, 2014 ). Cyclo-RGD truncated polymeric nanoconstructs with dendrimeric templates for targeted HDAC4 gene silencing inhibited the progression of renal fibrosis in an STZ-induced DN model ( Raval et al, 2021 ). Silencing of HDAC9 attenuates glomerulosclerosis, inflammatory cytokine release, podocyte apoptosis and injury in db/db mice ( Liu et al, 2016 ).…”

Dendronized Polymeric Biomaterial for Loading, Stabilization, and Targeted Cytosolic Delivery of microRNA in Cancer Cells

“…The MS/MS parameters for positive mode scan are 1000 V Nozzle voltage, 3000 V capillary voltage, 160 V fragmentor voltage, and 65 V skimmer. 17 2.5. Effect of Deprotected D2 Dendron Grafting on dCHA on RBCs, Hemolysis, Particle Size, and Zeta Potential.…”

“…For the evaluation of complex formation, a gel retardation assay was performed. 17 Control samples containing anti-miR21 (10,20,30,40, and 50 pmol) were also prepared. All the control samples were adjusted to the same volume (25 μL) as the sample with nuclease-free water.…”

“…The N-Boc-deprotected D2 dendron was conjugated with HA by the EDC coupling reaction as specified in the Thermo Scientific protocol. 17 Briefly, HA (20 mg, 0.0004 mmol) was reacted with EDC (10.543 mg, 0.055 mmol) and N-Boc-deprotected D2 dendron (37 mg, 0.06 mmol) at room temperature (400 rpm; 2 h). The reaction was performed in a conjugation buffer comprising 0.1 M MES (pH 4.5−5; 4 mL).…”

Histone Acetylation and Modifiers in Renal Fibrosis

“…In cultured renal epithelial cells, MC1568 treatment and siRNA-mediated HDAC4 silencing also inhibited expression of several profibrotic markers, including α-SMA, fibronectin and collagen 1, as well as phosphorylation of Smad3 and NF-κB ( Xiong et al, 2019 ). Another study showed that HDAC4 silencing attenuated glomerular fibrosis and inhibited expression of the aformentioned fibrotic proteins in diabetic mice ( Raval et al, 2021 ). Thus, we suggest that among class IIa HDACs, HDAC4 may play a predominant role in promoting renal fibrosis.…”