Multifunctional Lipid Nanoparticles for Protein Kinase N3 shRNA Delivery and Prostate Cancer Therapy

Abstract

Protein kinase N3 (PKN3), by virtue of its abnormal expression in prostate cells, has been widely used as a target of RNAi (shRNA, siRNA, miRNA) therapy. The major challenges of PKN3 RNAi therapy lie in how to design effective interference sequences and delivery systems. Herein, new PKN3 shRNA sequences (shPKN3-2459 and shPKN3-3357) were designed, and bioreducible, biodegradable, ionizable lipid-based nanoparticles were developed for shPKN3 delivery. First, an ionizable lipid (DDA-SS-DMA) bridged with disulfide bond and ester bonds was synthesized by a three-step reaction and confirmed by MS, 1 H NMR, and 13 C NMR. The ionizable lipid was mixed with cholesterol, DSPC, PEG-lipid, and shPKN3 by a microfluidic mixer to prepare lipid nanoparticles (LNP−shPKN3) which were characterized by DLS and TEM. Afterward, the pH and glutathione (GSH)-responsiveness of the DDA-SS-DMA based LNP delivery system were investigated by lysosome escape and gel electrophoresis assays. Compared with the commercial transfection reagent Lipo2000, the DDA-SS-DMA based delivery system showed higher transfection efficiency and lower toxicity. Western blot analysis, invasion tests, and migration assays were performed to evaluate the silencing effect of shPKN3 in vitro. In in vivo studies, high tumor suppression (65.8%) and treatment safety were evident in the LNP−shPKN3-2459 treatment group. Taken together, the DDA-SS-DMA based delivery system encapsulating shPKN3-2459 showed significant antitumor efficacy and might be a promising formulation for the treatment of prostate cancer.

Nanomedicine for Combination Urologic Cancer Immunotherapy

“…Wang et al designed ionizable liposomes to carry protein kinase N3 (PKN3), which is aberrantly expressed in prostate cancer cells. The nanoparticles vigorously suppressed tumor growth (65.8%) and improved therapeutic safety [ 116 ]. Similarly, nanoparticles containing PTT/PDT have been used in prostate cancer.…”

MicroRNA-nanoparticles against cancer: Opportunities and challenges for personalized medicine

“… 103 Wang and colleagues silenced protein kinase N3 (PKN3), strongly expressed in prostate cancer cells, by developing a bioreducible, biodegradable, and ionizable lipid-based NP encapsulating shPKN3. 104 This innovative method counteracted cancer progression and was a more effective and less toxic transfectant agent increasing in vivo tumor suppression. 104 Recent evidence showed the antitumor action of icaritin loading poly (lactic-co-glycolic acid) (PLGA@Icaritin) NPs in in vivo models of gastric cancer.…”

“… 104 This innovative method counteracted cancer progression and was a more effective and less toxic transfectant agent increasing in vivo tumor suppression. 104 Recent evidence showed the antitumor action of icaritin loading poly (lactic-co-glycolic acid) (PLGA@Icaritin) NPs in in vivo models of gastric cancer. Particularly, PLGA@Icaritin NPs stimulated ROS production, altered mitochondrial membrane potential lowering, and enhanced immune system response against tumor cells, thus impairing cancer progression.…”

Ionizable drug delivery systems for efficient and selective gene therapy

“…The creation of obstructing sequences and delivery mechanisms is a significant current problem in PKN3 RNAi treatment. Wang et al [ 174 ] synthesized a multifunctional ILNP (DDA-SS-DMA) that integrated ester bonds and disulfide bonds for PKN3 shRNA delivery and prostate cancer therapy. In particular, shPKN3-3357 and shPKN3-2459 were also developed as novel PKN3 shRNA sequences.…”

“…The results showed that ILNP containing the ionizable cationic lipid (DLin-KC2-DMA) effectively silenced the AR gene in both wild-type AR-expressing cells (LAPC-4) and LNCaP xenograft tumors. This study also suggested that strategies to improve targeting of ILNP AR-siRNA systems to prostate cancer cells may be more effective in knocking down AR and treating advanced prostate cancer [ 174 , 175 ].…”