Novel L-RNA Aptamer Controls <i>APP</i> Gene Expression in Cells by Targeting RNA G-Quadruplex Structure

Zhao, Haizhou; Wong, Hei Yuen; Ji, Dong; Lyu, Kaixin; Kwok, Chun Kit

doi:10.1021/acsami.2c06390

Cited by 8 publications

(3 citation statements)

References 55 publications

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“…To further corroborate these findings, we utilized antisense oligonucleotides (ASOs) against the FPGS 3′UTR GQs. ASOs have been recently utilized for sequence-specific inhibition of RNA GQ folding, thus reducing protein translation [ 97 – 99 ]. We here employed the same technique with the aim to abrogate the FA-induced cell protrusion localization of cFPGS mRNA.…”

Section: Resultsmentioning

confidence: 99%

Folylpolyglutamate synthetase mRNA G-quadruplexes regulate its cell protrusion localization and enhance a cancer cell invasive phenotype upon folate repletion

et al. 2023

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Background Folates are crucial for the biosynthesis of nucleotides and amino acids, essential for cell proliferation and development. Folate deficiency induces DNA damage, developmental defects, and tumorigenicity. The obligatory enzyme folylpolyglutamate synthetase (FPGS) mediates intracellular folate retention via cytosolic and mitochondrial folate polyglutamylation. Our previous paper demonstrated the association of the cytosolic FPGS (cFPGS) with the cytoskeleton and various cell protrusion proteins. Based on these recent findings, the aim of the current study was to investigate the potential role of cFPGS at cell protrusions. Results Here we uncovered a central role for two G-quadruplex (GQ) motifs in the 3′UTR of FPGS mediating the localization of cFPGS mRNA and protein at cell protrusions. Using the MBSV6-loop reporter system and fluorescence microscopy, we demonstrate that following folate deprivation, cFPGS mRNA is retained in the endoplasmic reticulum, whereas upon 15 min of folate repletion, this mRNA is rapidly translocated to cell protrusions in a 3′UTR- and actin-dependent manner. The actin dependency of this folate-induced mRNA translocation is shown by treatment with Latrunculin B and inhibitors of the Ras homolog family member A (RhoA) pathway. Upon folate repletion, the FPGS 3′UTR GQs induce an amoeboid/mesenchymal hybrid cell phenotype during migration and invasion through a collagen gel matrix. Targeted disruption of the 3′UTR GQ motifs by introducing point mutations or masking them by antisense oligonucleotides abrogated cell protrusion targeting of cFPGS mRNA. Conclusions Collectively, the GQ motifs within the 3′UTR of FPGS regulate its transcript and protein localization at cell protrusions in response to a folate cue, inducing cancer cell invasive phenotype. These novel findings suggest that the 3′UTR GQ motifs of FPGS constitute an attractive druggable target aimed at inhibition of cancer invasion and metastasis.

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Section: Resultsmentioning

confidence: 99%

Folylpolyglutamate synthetase mRNA G-quadruplexes regulate its cell protrusion localization and enhance a cancer cell invasive phenotype upon folate repletion

et al. 2023

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“…L‐RNA aptamers (“mirror image” of its D‐RNA counterparts) are resistant to nuclease degradation and can interact with specific rG4 through structural recognition [9] . Recently, our group reported the first few L‐RNA aptamers, [10] for the specific binding to rG4s of interest, providing a new rG4 toolset for visualizing rG4s in fixed cells, [10d] controlling rG4‐protein interactions, and regulating rG4‐linked gene activity [10a] . L‐Apt.4‐1c is the shortest (25 nt) L‐aptamer ever developed, that preferentially binds to rG4s over dG4s and non‐G4s [10c] (Figure 1A).…”

Section: Introductionmentioning

confidence: 99%

Rational Design of L‐RNA Aptamer‐Peptide Conjugate for Efficient Cell Uptake and G‐quadruplex‐Mediated Gene Control

Zhang,

Nie,

Chi‐Kong Lau

et al. 2024

Angew Chem Int Ed

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RNA G‐quadruplexes (D‐rG4s) are prevalent in the transcriptome and play crucial regulatory roles in various biological processes. Recently, L‐RNA aptamers have been reported to recognize functional rG4s with a strong binding affinity and specificity. However, owing to the poor cell penetration capacity of L‐RNA aptamers, their biological applications are currently limited. Herein, we rationally design an L‐RNA aptamer‐peptide conjugate, Tamra_Ahx_R8_L‐Apt.4‐1c, which can efficiently translocate into the cytosol and target the rG4 of interest. Notably, we demonstrate diverse regulatory roles of Tamra_Ahx_R8_L‐Apt.4‐1c on rG4 motif present in different regions of mRNAs and further expand the application in different cell lines. Our novel and biocompatible conjugate enhances the cellular uptake of the L‐RNA aptamer, and our robust strategy enables non‐canonical RNA structures to be targeted by L‐RNA aptamers for gene control in cells.

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“…Aptamers are single-stranded oligonucleotides with unique three dimensional structures that can recognize specific targets, such as proteins, nucleic acids or cells, with high affinity [ 17 – 19 ]. Moreover, aptamers have several advantages such as synthetic accessibility, low toxicity, flexible chemical modifications and a lack of immunogenicity [ 20 , 21 ].…”

Section: Introductionmentioning

confidence: 99%

Natural killer cells modified with a Gpc3 aptamer enhance adoptive immunotherapy for hepatocellular carcinoma

Zheng,

Lai,

Wang

et al. 2023

Discov Onc

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Introduction Natural killer cells can attack cancer cells without prior sensitization, but their clinical benefit is limited owing to their poor selectivity that is caused by the lack of specific receptors to target tumor cells. In this study, we aimed to endow NK cells with the ability to specifically target glypican-3+ tumor cells without producing cell damage or genetic alterations, and further evaluated their therapeutic efficiency. Methods NK cells were modified with a Gpc3 DNA aptamer on the cell surface via metabolic glycoengineering to endow NK cells with specific targeting ability. Then, the G-NK cells were evaluated for their specific targeting properties, cytotoxicity and secretion of cytokines in vitro. Finally, we investigated the therapeutic efficiency of G-NK cells against glypican-3+ tumor cells in vivo. Results Compared with NK cells modified with a random aptamer mutation and unmodified NK cells, G-NK cells induced significant apoptosis/necrosis of GPC3+ tumor cells and secreted cytokines to preserve the intense cytotoxic activities. Moreover, G-NK cells significantly suppressed tumor growth in HepG2 tumor-bearing mice due to the enhanced enrichment of G-NK cells at the tumor site. Conclusions The proposed strategy endows NK cells with a tumor-specific targeting ability to enhance adoptive therapeutic efficiency in GPC3+ hepatocellular carcinoma.

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Novel L-RNA Aptamer Controls APP Gene Expression in Cells by Targeting RNA G-Quadruplex Structure

Cited by 8 publications

References 55 publications

Folylpolyglutamate synthetase mRNA G-quadruplexes regulate its cell protrusion localization and enhance a cancer cell invasive phenotype upon folate repletion

Folylpolyglutamate synthetase mRNA G-quadruplexes regulate its cell protrusion localization and enhance a cancer cell invasive phenotype upon folate repletion

Rational Design of L‐RNA Aptamer‐Peptide Conjugate for Efficient Cell Uptake and G‐quadruplex‐Mediated Gene Control

Natural killer cells modified with a Gpc3 aptamer enhance adoptive immunotherapy for hepatocellular carcinoma

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