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Updated: Oct 26, 2025
Preparation of Neutrally-charged, pH-responsive Polymeric Nanoparticles for Cytosolic siRNA Delivery
Published on: May 2, 2019
Antibody-siRNA conjugates (ARCs) using multifunctional peptide as a tumor enzyme cleavable linker mediated effective
Zhili Yu1, Xiaojuan Zhang2, Xing Pei2
1Research Center of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China.
Abstract:
The tissue-specific targeted delivery and efficient cellular uptake of siRNAs are the main obstacles to their clinical application. Antibody-siRNA-conjugates (ARCs) can deliver siRNA by exploiting the targeting property of antibodies like antibody-drug conjugates (ADCs). However, the effective conjugation of antibodies and siRNAs and the release of siRNAs specifically at target sites have posed challenges to the development of ARCs. In this study, the successful conjugation of antibodies and siRNAs was achieved using a multifunctional peptide as a linker, composed of a cell-penetrating peptide (CPP) and a substrate peptide (SP), which is highly expressed in solid tumors. The resulting antibody-multifunctional peptide (SP-CPP)-siRNA system delivered the siRNA to target tumor cells by the specific binding of the antibody. Once the enzymes on the tumor cell surface hydrolyzed the substrate peptide linker, siRNA-CPP was released from ARCs. The released siRNA-CPP entered the targeted cells via the cellular penetrating ability of CPP, resulting in improved siRNA-mediated gene silencing efficiency, verified both in vitro and in vivo. After intravenous administration, the designed ARCs achieved approximately 66.7% EGFP (Enhanced Green Fluorescent Protein) downregulation efficiency in nude mice xenografted with the HCT116-EGFP tumor model. The proposed system provides a prospective choice for ARC production and the safe and efficient delivery of siRNAs.
Insights
This study introduces a novel antibody-siRNA-conjugate (ARC) system using a multifunctional peptide linker for enhanced tumor cell delivery and gene silencing. The innovative ARC design improves siRNA delivery and therapeutic efficacy, overcoming key clinical application hurdles.
Area of Science:
- Biotechnology
- Molecular Biology
- Drug Delivery Systems
Background:
- Targeted delivery and cellular uptake of small interfering RNAs (siRNAs) are critical challenges for their clinical use.
- Antibody-siRNA-conjugates (ARCs) offer a promising strategy for siRNA delivery, mirroring antibody-drug conjugate (ADC) technology.
- Existing ARC development faces hurdles in antibody-siRNA conjugation and site-specific siRNA release.
Purpose of the Study:
- To develop a novel antibody-siRNA-conjugate (ARC) system for efficient and targeted siRNA delivery to solid tumors.
- To overcome the limitations of antibody-siRNA conjugation and achieve specific siRNA release at target sites.
- To enhance siRNA-mediated gene silencing efficiency both in vitro and in vivo.
Main Methods:
- Conjugation of antibodies and siRNAs using a multifunctional peptide linker containing a cell-penetrating peptide (CPP) and a tumor-expressed substrate peptide (SP).
- Development of an antibody-multifunctional peptide (SP-CPP)-siRNA system for targeted delivery via antibody binding.
- In vitro and in vivo validation of siRNA release, cellular uptake, and gene silencing efficacy.
Main Results:
- Successful conjugation of antibodies and siRNAs was achieved using the SP-CPP linker.
- The ARC system demonstrated targeted delivery to tumor cells, followed by enzyme-mediated release of siRNA-CPP.
- In vivo studies in a HCT116-EGFP tumor model showed approximately 66.7% EGFP downregulation efficiency after intravenous administration.
- Enhanced siRNA-mediated gene silencing was confirmed both in vitro and in vivo.
Conclusions:
- The developed antibody-multifunctional peptide (SP-CPP)-siRNA system represents a viable approach for ARC production.
- This system facilitates safe and efficient siRNA delivery, addressing key obstacles to clinical application.
- The study provides a promising strategy for improving the therapeutic potential of siRNA-based treatments.
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