Oligonucleotide Molecular Sprinkler for Intracellular Detection and Spontaneous Regulation of mRNA for Theranostics of Scar Fibroblasts

Abstract

Early diagnosis and timely intervention are key for the successful treatment of skin diseases like abnormal scars. This study introduces a nucleic‐acid‐based probe (i.e., molecular sprinkler) for the diagnosis and spontaneous regulation of the abnormal expression of fibrosis‐related mRNA in scar‐derived skin fibroblasts. Using mRNA encoding connective tissue growth factor (CTGF) as the model gene, a probe with three oligonucleotides is constructed, including a recognition sequence complementary to the CTGF mRNA, a siRNA against transforming growth factor receptor I (TGFβRI) as the CTGF mRNA suppressor, and a connecting sequence. The probe can detect CTGF mRNA with a limit of 10 × 10−9 m and distinguishes scar fibroblasts from normal ones in both 2D and 3D environments. Two days after transfection, the siRNA released from the probe reduces the expression of TGFβRI and, consequently, decreases the cellular expression of CTGF mRNA (up to 70%). This dual‐role probe presents opportunities to monitor the TGF‐ β signaling pathway, screen for drugs that target the CTGF pathway, and determine the role of inhibition of the CTGF pathway in therapeutic efficacy.

Advances in the Formulations of Microneedles for Manifold Biomedical Applications

“…Differing from the above biodegradable MNs that permit sustained drug release, dissolving MNs offer a burst release profile. The representative polymers are hyaluronic acid (HA),8a,40 carboxymethlycellulose (CMC),8b,41 polyacrylic acid (PAA),8d PVA, PVP, sucrose, starch, trehalose,41a DNA, and their mixtures 41a,46. McCrudden and co‐workers designed a dissolving MN patch for the transdermal delivery of painkiller, ibuprofen by screening a range of food and drug administration (FDA) regulated polymers and their combinations to maximize the drug loading and mechanical properties.…”

“…Xue and co‐workers developed an anti‐scar MN patch using PEGDA and the anti‐metabolite drug, 5‐fluorouracil (5‐FU) 9g. Zheng et al further applied a dissolving HA MN patch to deliver the dual‐function nucleic acid probe (i.e., molecular sprinkler) for the detection and suppression of abnormal mRNA expression in scar‐derived fibroblasts (Figure C,D) . The dissolvable MNs rapidly delivered the probe into 3D cultured hypertrophic scar fibroblasts (HSF) model and allowed the detection and modulation of scar‐related biomarker, connective tissue growth factor in HSF .…”

“…D) Downregulation of cellular CTGF mRNA through the suppression of TGFβRI expression. Reproduced with permission . Copyright 2018, Wiley‐VCH.…”

Hydrogel-Based Technologies for the Diagnosis of Skin Pathology

“…Given the pairwise hybridization nature of NA (purines-pyrimidines: A-T/U and C-G), it is unsurprising that most NA sensors rely on complementary NAs (natural or backbone-modified oligonucleotides) tagged with the inducible FRET fluorescence system (i.e., fluorophore-quencher/fluorophore-fluorophore) [94]. Notable examples include the hairpin-shaped molecular beacons, DNAzymes, detachable reporter flares, as well as their formulations on nanoparticles with quenching capacity (e.g., Au nanospheres/nanorods and graphene oxide nanosheets; Figure 4A-C) [95][96][97]. Interestingly, certain probes such as spherical nucleic acids-based ones are capable of entering exosomal vesicles to bind and directly report the presence of mRNA/miRNA within it [98].…”

“…nanoparticles with quenching capacity (e.g., Au nanospheres/nanorods and graphene oxide nanosheets; Figure 4A-C) [95][96][97]. Interestingly, certain probes such as spherical nucleic acids-based ones are capable of entering exosomal vesicles to bind and directly report the presence of mRNA/miRNA within it [98].…”