Targeted delivery of siRNA to activated T cells via transferrin-polyethylenimine (Tf-PEI) as a potential therapy of asthma

Xie, Yuran; Kim, Na-Hyung; Nadithe, Venkatareddy; Schalk, Dana; Thakur, Archana; Kılıç, Ayşe; Lum, Lawrence G.; Bassett, D. J. P.; Merkel, Olivia M.

doi:10.1016/j.jconrel.2016.03.029

Cited by 100 publications

(101 citation statements)

References 55 publications

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“…[30] To investigate the ability of VIPER to deliver siRNA via pulmonary delivery, a murine model for pulmonary delivery was used. [7] Mice were intratracheally administered CP or VIPER polyplexes that were made with either TYE™-563 or pHrodo™ labeled siRNA at N/P 8 (Figure 8A). The pHrodo™ dye is a commercially available fluorescent pH indicator that has been reported to dramatically increase in fluorescence as the acidity of the environment changes.…”

Section: Resultsmentioning

confidence: 99%

“…[6] Numerous studies have identified endosomal release to be one of the key factors contributing to successful transfection efficiency in cells treated with polymeric carriers of nucleic acid-based drugs. [7] In order to address the issue of endosomal entrapment, various mechanisms have been utilized to improve endosomal release of synthetic polymer carriers. [8] The use of polymers with buffering capabilities has been hypothesized to lead to the rupture of the endosomal membrane, but requires concentrations of polymer that are difficult to achieve in vivo .…”

Section: Introductionmentioning

confidence: 99%

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In vitro and in vivo delivery of siRNA via VIPER polymer system to lung cells

Feldmann

Cheng

Kandil

et al. 2018

Journal of Controlled Release

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The block copolymer VIPER (virus-inspired polymer for endosomal release) has been reported to be a promising novel delivery system of DNA plasmids both in vitro and in vivo. VIPER is comprised of a polycation segment for condensation of nucleic acids as well as a pH-sensitive segment that exposes the membrane lytic peptide melittin in acidic environments to facilitate endosomal escape. The objective of this study was to investigate VIPER/siRNA polyplex characteristics, and compare their in vitro and in vivo performance with commercially available transfection reagents and a control version of VIPER lacking melittin. VIPER/siRNA polyplexes were formulated and characterized at various charge ratios and shown to be efficiently internalized in cultured cells. Target mRNA knockdown was confirmed by both flow cytometry and qRT-PCR and the kinetics of knockdown was monitored by live cell spinning disk microscopy, revealing knockdown starting by 4 hours post-delivery. Intratracheal instillation of VIPER particles formulated with sequence specific siRNA to the lung of mice resulted in a significantly more efficient knockdown of GAPDH compared to treatment with VIPER particles formulated with scrambled sequence siRNA. We also demonstrated using pH-sensitive labels that VIPER particles experience less acidic environments compared to control polyplexes. In summary, VIPER/siRNA polyplexes efficiently deliver siRNA in vivo resulting in robust gene silencing (>75% knockdown) within the lung.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

In vitro and in vivo delivery of siRNA via VIPER polymer system to lung cells

Feldmann

Cheng

Kandil

et al. 2018

Journal of Controlled Release

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“…We have attempted to perform genomic modification of the two target lncRNAs in murine primary Tm cells with small-interfering RNA. However, primary T lymphocytes are generally resistant to non-viral vector-based transfection methods [38]. Significant time and effort is still needed to solve this problem.…”

Section: Discussionmentioning

confidence: 99%

The lncRNAs involved in mouse airway allergic inflammation following induced pluripotent stem cell-mesenchymal stem cell treatment

Wang

Fan

et al. 2017

Stem Cell Res Ther

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BackgroundWe have previously reported that induced pluripotent stem cell (iPSC)-mesenchymal stem cells (MSCs) alleviated asthma inflammation in mice. Long noncoding RNAs (lncRNAs) were recently reported as being involved in the immune responses. However, whether lncRNAs are associated with iPSC-MSC immunomodulation in allergic inflammation is still unclear.MethodsMice were induced into an asthmatic state and received treatment consisting of iPSC-MSCs. Memory T cells isolated from sensitized mice were challenged and co-cultured with iPSC-MSCs in vitro. Total RNA from the lungs and separated T cells were processed with an lncRNA/mRNA microarray. A series of bioinformatics technologies were used to screen the target lncRNAs.ResultsiPSC-MSCs significantly prevented asthma inflammation and decreased the Th2 cytokine levels. Over 1300 lncRNAs were differentially expressed after the induction of asthma, and 846 or 4176 lncRNAs were differentially expressed with iPSC-MSC treatment in mice or in vitro, respectively. After overlapping the differentially expressed lncRNAs produced in a similar manner in mice and in vitro, 23 lncRNAs and 96 mRNAs were selected, in which 58 protein-coding genes were predicted to be potential targets of the 23 lncRNAs. Furthermore, using a series of bioinformatics technologies, 9 lncRNAs co-expressed with the most differentially expressed mRNAs, which were enriched in terms of the immune response, were screened out via Pearson’s correlation coefficient with mRNAs that were involved with inflammatory cytokines and receptors. lncRNAs MM9LINCRNAEXON12105+ and AK089315 were finally emphasized via quantitative real-time PCR validation.ConclusionsOur results suggested that aberrant lncRNA profiles were present after asthma induction and iPSC-MSC treatment, suggesting potential targets of allergic inflammation and iPSC-MSC-mediated immunomodulation.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0456-3) contains supplementary material, which is available to authorized users.

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“…19 PEI (1 mg/mL) was reacted with SPDP or PEG4-SPDP (20 mM) in DMSO stirring at room temperature overnight (1:10 molar ratio relative to amines present). The intermediate products were purified by ultrafiltration using Amicon Ultra 4 mL filters (MWCO 3000) and Vivaspin centrifugal ultrafilters (10,000 MWCO) as described.…”

Section: Methodsmentioning

confidence: 99%

Post-Transcriptional Regulation of the GASC1 Oncogene with Active Tumor-Targeted siRNA-Nanoparticles

et al. 2016

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Basal-like breast cancer (BLBC) accounts for the most aggressive types of breast cancer, marked by high rates of relapse and poor prognoses and with no effective clinical therapy yet. Therefore, investigation of new targets and treatment strategies is more than necessary. Here, we identified a receptor that can be targeted in BLBC for efficient and specific siRNA mediated gene knockdown of therapeutically relevant genes such as the histone demethylase GASC1, which is involved in multiple signaling pathways leading to tumorigenesis. Breast cancer and healthy breast cell lines were compared regarding transferrin receptor (TfR) expression via flow cytometry and transferrin binding assays. Nanobioconjugates made of low molecular weight polyethylenimine (LMW-PEI) and transferrin (Tf) were synthesized to contain a bioreducible disulfide bond. siRNA complexation was characterized by condensation assays and dynamic light scattering. Cytotoxicity, transfection efficiency, and the targeting specificity of the conjugates were investigated in TfR positive and negative healthy breast and breast cancer cell lines by flow cytometry, confocal microscopy, RT-PCR, and Western blot. Breast cancer cell lines revealed a significantly higher TfR expression than healthy breast cells. The conjugates efficiently condensed siRNA into particles with 45 nm size at low polymer concentrations, showed no apparent toxicity on different breast cancer cell lines, and had significantly greater transfection and gene knockdown activity on mRNA and protein levels than PEI/siRNA leading to targeted and therapeutic growth inhibition post GASC1 knockdown. The synthesized nanobioconjugates improved the efficiency of gene transfer and targeting specificity in transferrin receptor positive cells but not in cells with basal receptor expression. Therefore, these materials in combination with our newly identified siRNA sequences are promising candidates for therapeutic targeting of hard-to-treat BLBC and are currently further investigated regarding in vivo targeting efficacy and biocompatibility.

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Targeted delivery of siRNA to activated T cells via transferrin-polyethylenimine (Tf-PEI) as a potential therapy of asthma

Cited by 100 publications

References 55 publications

In vitro and in vivo delivery of siRNA via VIPER polymer system to lung cells

In vitro and in vivo delivery of siRNA via VIPER polymer system to lung cells

The lncRNAs involved in mouse airway allergic inflammation following induced pluripotent stem cell-mesenchymal stem cell treatment

Post-Transcriptional Regulation of the GASC1 Oncogene with Active Tumor-Targeted siRNA-Nanoparticles

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