Method and its Composition for encapsulation, stabilization, and delivery of siRNA in Anionic polymeric nanoplex: An In vitro- In vivo Assessment

Nidhi Raval,Kiran Kalia,Hardi Jogi,Rakesh K Tekade,Piyush Gondaliya

doi:10.1038/s41598-019-52390-4

Nidhi Raval, Kiran Kalia + Show 3 more

Open Access

https://doi.org/10.1038/s41598-019-52390-4

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Abstract

Small interfering RNA (siRNA) are synthetic RNA duplex designed to specifically knockdown the abnormal gene to treat a disease at cellular and molecular levels. In spite of their high potency, specificity, and therapeutic potential, the full-fledged utility of siRNA is predominantly limited to in vitro set-up. Till date, Onpattro is the only USFDA approved siRNA therapeutics available in the clinic. The lack of a reliable in vivo siRNA delivery carrier remains a foremost obstacle towards the clinical translation of siRNA therapeutics. To address the obstacles associated with siRNA delivery, we tested a dendrimer-templated polymeric approach involving a USFDA approved carrier (albumin) for in vitro as well as in vivo delivery of siRNA. The developed approach is simple in application, enhances the serum stability, avoids in vivo RNase-degradation and mediates cytosolic delivery of siRNA following the endosomal escape process. The successful in vitro and in vivo delivery of siRNA, as well as targeted gene knockdown potential, was demonstrated by HDAC4 inhibition in vitro diabetic nephropathy (DN) podocyte model as well as in vivo DN C57BL/6 mice model. The developed approach has been tested using HDAC4 siRNA as a model therapeutics, while the application can also be extended to other gene therapeutics including micro RNA (miRNA), plasmids oligonucleotides, etc.

Highlights

Ribonucleic acid interference (RNAi) refers to a post-transcriptional gene silencing tool to neutralize or silence the pathological protein via activating RNA-induced silencing complex (RISC), endogenously
We evaluate and report it’s in vitro and in vivo HDAC4 gene silencing capability using podocytes as well as in diabetic nephropathy (DN) mouse model
The binding efficiency of Small interfering RNA (siRNA) with the polycationic dendrimer template was investigated by determining the band binding efficiency and band intensity of the siRNA (Fig. 1B,C)

Summary

Introduction

Ribonucleic acid interference (RNAi) refers to a post-transcriptional gene silencing tool to neutralize or silence the pathological protein via activating RNA-induced silencing complex (RISC), endogenously. Gene allows clinicians to treat a disease by administering RNAi therapeutics (such as small interfering RNA (siRNA) or micro RNA (miRNA) into a patient’s instead of using drugs or surgical interventions[1]. The siRNA delivery vector must find their way to escape from the endosomes/lysosomes compartment after entering inside the target cell to efficiently release the loaded siRNA in the cytosolic compartment[4]. Failure to overcome this barrier would significantly weaken or even totally eliminate the therapeutic effect of siRNA

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