Selective RNA interference and gene silencing using reactive oxygen species-responsive lipid nanoparticles

Liang, Chunjing; Chang, Jin; Jiang, Ying; Liu, Ji; Mao, Lanqun; Wang, Ming

doi:10.1039/c9cc04517a

Cited by 23 publications

(27 citation statements)

References 33 publications

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“…The siRNAs were transfected using Interferin (Polyplus #409) at a final concentration of 30 nM. Two siRNA couples, siTRIM47 #1 and #2 targeting human TRIM47 were used 34 and a non-specific siRNA as negative control (Eurogentec ® ). SiRNA-depletion was then resolved by sodium dodecyl sulphate–polyacrylamide gel electrophoresis using TRIM47 antibody (Thermofisher Scientific, #PA5-50892).…”

Section: Methodsmentioning

confidence: 99%

Gene-mapping study of extremes of cerebral small vessel disease reveals TRIM47 as a strong candidate

Mishra

Duplàa

Vojinović

et al. 2022

Brain

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Cerebral small vessel disease is a leading cause of stroke and a major contributor to cognitive decline and dementia, but our understanding of specific genes underlying the cause of sporadic cerebral small vessel disease is limited. We report a genome-wide association study and a whole-exome association study on a composite extreme phenotype of cerebral small vessel disease derived from its most common MRI features: white matter hyperintensities and lacunes. Seventeen population-based cohorts of older persons with MRI measurements and genome-wide genotyping (n = 41 326), whole-exome sequencing (n = 15 965), or exome chip (n = 5249) data contributed 13 776 and 7079 extreme small vessel disease samples for the genome-wide association study and whole-exome association study, respectively. The genome-wide association study identified significant association of common variants in 11 loci with extreme small vessel disease, of which the chr12q24.11 locus was not previously reported to be associated with any MRI marker of cerebral small vessel disease. The whole-exome association study identified significant associations of extreme small vessel disease with common variants in the 5′ UTR region of EFEMP1 (chr2p16.1) and one probably damaging common missense variant in TRIM47 (chr17q25.1). Mendelian randomization supports the causal association of extensive small vessel disease severity with increased risk of stroke and Alzheimer’s disease. Combined evidence from summary-based Mendelian randomization studies and profiling of human loss-of-function allele carriers showed an inverse relation between TRIM47 expression in the brain and blood vessels and extensive small vessel disease severity. We observed significant enrichment of Trim47 in isolated brain vessel preparations compared to total brain fraction in mice, in line with the literature showing Trim47 enrichment in brain endothelial cells at single cell level. Functional evaluation of TRIM47 by small interfering RNAs-mediated knockdown in human brain endothelial cells showed increased endothelial permeability, an important hallmark of cerebral small vessel disease pathology. Overall, our comprehensive gene-mapping study and preliminary functional evaluation suggests a putative role of TRIM47 in the pathophysiology of cerebral small vessel disease, making it an important candidate for extensive in vivo explorations and future translational work.

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Section: Methodsmentioning

confidence: 99%

Gene-mapping study of extremes of cerebral small vessel disease reveals TRIM47 as a strong candidate

Mishra

Duplàa

Vojinović

et al. 2022

Brain

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“…9,32 More recently, we demonstrated that integrating reactive oxygen species (ROS)-cleavable thioketal moiety into lipid nanoparticles promotes intracellular lipid degradation and cargo release in cancer cells for an enhanced gene silencing. 33 We hypothesized that such a ROS-enhanced delivery strategy could be similarly adapted for delivering RNase A-QPN to promote protein release inside cancer cells, enabling NQO1-catalyzed RNase A-QPN activation. Thus, we synthesized a library containing 12 ROS-responsive lipids (Figure S6) via the Michael addition between aliphatic amine and acrylate (Scheme S2, Supporting Information).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…However, the conditional control of RNase A-QPN activity by tumor-cell-specific NQO1 is greatly prevented by the low efficiency of the protein in entering the cells. − It is therefore of utmost importance to deliver the protein conjugate into tumor cells to enable NQO1 activation and targeted cancer therapy. Recently, we used a combinatorial strategy to synthesize a library of cationic lipids, and to screen effective protein delivery vehicles by self-assembling protein–lipid nanoparticles. , More recently, we demonstrated that integrating reactive oxygen species (ROS)-cleavable thioketal moiety into lipid nanoparticles promotes intracellular lipid degradation and cargo release in cancer cells for an enhanced gene silencing . We hypothesized that such a ROS-enhanced delivery strategy could be similarly adapted for delivering RNase A-QPN to promote protein release inside cancer cells, enabling NQO1-catalyzed RNase A-QPN activation.…”

Section: Results and Discussionmentioning

confidence: 99%

Enzyme-Instructed Activation of Pro-protein Therapeutics In Vivo

et al. 2019

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The selective and temporal control of protein activity in living cells provides a powerful tool to manipulate cellular function and to develop pro-protein therapeutics (PPT) for targeted therapy. In this work, we reported a facile but general chemical approach to design PPT by modulating protein activity in response to endogenous enzyme of disease cells, and its potential for targeted cancer therapy. We demonstrated that the chemical modification of a protein with quinone propionic acid (QPN), a ligand that could be reduced by tumor-cell-specific NAD(P)H dehydrogenase [quinone] 1 (NQO1), was reversible in the presence of NQO1. Importantly, the QPN-modified cytochrome c (Cyt c-QPN) and ribonuclease A (RNase A-QPN) showed NQO1-regulated protein activity in a highly selective manner. Furthermore, the intracellular delivery of RNase A-QPN using a novel type of lipid-based nanoparticles, and subsequent protein activation by cellular NQO1, selectively inhibit cancer cell growth in vitro and effectively suppress tumor growth in vivo. We believe that our approach increases the number of potentially useful chemical tools for reversibly controlling the structure and function of protein using a disease-cell-specific enzyme, opening opportunities in the study of dynamic biological processes and developing precise protein therapeutics.

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“…[33][34][35][36][37] Herein, we used a cationic lipid, EC16-80, which had recently been developed for effective protein delivery. EC16-80 was screened from a combinatorial library of lipid nanoparticles, 23,38,39 for simultaneous delivery of RNase A-NBC and β-Lap into cells (Figure 1b). In the lipid nanoparticle formulation, RNase A-NBC was encapsulated mainly via electrostatic interaction with EC16-80, whereas β-Lap was entrapped in the hydrophobic layer of the lipid nanoparticles (Figure 1b).…”

Section: Enzymatic Activation Of Rnase A-nbc In Living Cellsmentioning

confidence: 99%

In Vivo Activation of Pro-Protein Therapeutics via Chemically Engineered Enzyme Cascade Reaction

et al. 2021

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Selective and temporal control over protein activity is of great importance for the advancement of the protein of interest into precise molecular medicine. Simply installing synthetic ligands to proteins for activity regulation, however, is often obscured by either nonspecificity or insufficient efficiency. This study reports a chemical approach in which enzymatic cascade reactions were designed for selective activation of pro-protein both in vitro and in vivo. Specifically, the system consisted of aromatic boronic-acid-modified nanoparticles, reactive oxygen species (ROS)-responsive pro-protein (RNase A-NBC), a small molecule drug, β-Lapachone (β-Lap), and strategically screened synthetic lipids, required for the assembly of the nanocomplexes. Once target-delivered into tumor cells, the reduction of β-Lap produces massive H 2 O 2 in response to NAD (P)H quinone oxidoreductase 1 (NQO1), a tumor-specific enzyme, which triggers further induction by selective chemical modification of ROS-responsive cytosolic protein ribonuclease A (RNase A)-NBC, thus, switching from "inert" pro-protein to active therapeutics, that ultimately prohibit tumor cell growth. Moreover, the designed enzymatic cascade activation of the pro-protein was effective in vivo, displaying superior therapeutic efficacy to either the pro-protein alone or the β-Lap via tumor-targeted delivery and the consequent suppression of the tumor growth. As both RNase A and β-Lap have been evaluated clinically as antitumor therapeutics, our chemical multi-step cascade methodology is, therefore, a promising strategy for selective modulation of pro-protein chemistry in the living system for fundamental investigations, favorable toward potential anticancer applications.

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Selective RNA interference and gene silencing using reactive oxygen species-responsive lipid nanoparticles

Abstract: The integration of reactive oxygen species (ROS)-responsive thioketal group into lipids nanoparticles enables the efficient delivery of siRNA into cells, and selectively cancer cell gene expression silencing in response to the high level of intracellular ROS in cancer cells.

Cited by 23 publications

References 33 publications

Gene-mapping study of extremes of cerebral small vessel disease reveals TRIM47 as a strong candidate

Gene-mapping study of extremes of cerebral small vessel disease reveals TRIM47 as a strong candidate

Enzyme-Instructed Activation of Pro-protein Therapeutics In Vivo

In Vivo Activation of Pro-Protein Therapeutics via Chemically Engineered Enzyme Cascade Reaction

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