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Cited by 15RNA Interference and Gene DeliveryNanoparticle-Based Drug DeliveryMesenchymal stem cell research

Evaluating side effects of nanoparticle‐mediated siRNA delivery to mesenchymal stem cells using next generation sequencing and enrichment analysis

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Malcolm, Dominic W., Sorrells, Janet E., Van Twisk, Daniel, Thakar, Juilee, Benoit, Danielle S. W.·2016·Bioengineering & Translational Medicine
DOI 10.1002/btm2.10035PMC PMC5125403
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Abstract

RNA interference has immense potential to modulate cell functions. However, effective delivery of small interfering RNA (siRNA) while avoiding deleterious side effects has proven challenging. This study investigates both intended and unintended effects of diblock copolymer nanoparticle (NP) delivery of siRNA delivery to human mesenchymal stem cells (hMSC). Specifically, siRNA delivery was investigated at a range of NP‐siRNA:hMSC ratios with a focus on the effects of NP‐siRNA treatment on hMSC functions. Additionally, next generation RNA sequencing (RNAseq) was used with enrichment analysis to observe side effects in hMSC gene expression. Results show NP‐siRNA delivery is negatively correlated with hMSC density. However, higher NP‐siRNA:hMSC ratios increased cytotoxicity and decreased metabolic activity. hMSC proliferation was largely unaffected by NP‐siRNA treatment, except for a threefold reduction in hMSCs seeded at 4,000 cells/cm 2. Flow cytometry reveals that apoptosis is a function of NP‐siRNA treatment time and seeding density; ∼14% of the treated hMSCs seeded at 8,000 cells/cm 2 were annexin V+‐siRNA+ 24 hr after treatment, while 11% of the treated population was annexin V+‐siRNA−. RNAseq shows that NP‐siRNA treatment results in transcriptomic changes in hMSCs, while pathway analysis shows upregulation of apoptosis signaling and downregulation of metabolism, cell cycle, and DNA replication pathways, as corroborated by apoptosis, metabolism, and proliferation assays. Additionally, multiple innate immune signaling pathways such as toll‐like receptor, RIG‐I‐like receptor, and nuclear factor‐κB signaling pathways are upregulated. Furthermore, and consistent with traditional siRNA immune activation, cytokine–cytokine receptor signaling was also upregulated. Overall, this study provides insight into NP‐siRNA:hMSC ratios that are favorable for siRNA delivery. Moreover, NP‐siRNA delivery results in side effects across the hMSC transcriptome that suggest activation of the innate immunity that could alter MSC functions associated with their therapeutic potential.

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  • The overall effect and degree of enrichment was minimal compared to previously observed off-target effects with siRNA in rat livers and in cells.

    “the overall effect and degree of enrichment was minimal compared to previously observed off-target effects with siRNA in rat livers38 and in cells47”

    From A divalent siRNA chemical scaffold for potent and sustained modulation of gene expression throughout the central nervous system (2019) · cites this paper as [47]

    The Results section (page unavailable) reports pathways enriched after nanoparticle-mediated siRNA treatment, but does not say the overall effect or degree of enrichment was minimal compared with earlier off-target effects. Searches for the comparison with siRNA effects in rat livers and cells found no passage making it. (LLM, conf=0.73)

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References 66 · Cited by 15

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Authors
Malcolm, D. W., Sorrells, J. E., Van Twisk, D., Thakar, J., & Benoit, D. S. W.
Affiliations
  • University of Rochester Medicine (US)
  • University of Rochester (US)
  • Dept. of Microbiology and Immunology University of Rochester Rochester NY 14627
Corresponding author
Benoit, Danielle S. W.
ORCID
Venue
Bioengineering & Translational Medicine
Publisher
Wiley
Published
2016-06
Volume
1
Issue
2
Pages
193-206
ISSN
2380-6761 (ISSN-L)
DOI
10.1002/btm2.10035
PMC
PMC5125403
OpenAlex
W2523693109
Cited by
15
References
66
Citations by year
2024: 3 · 2023: 1 · 2021: 1 · 2019: 1 · 2018: 4 · 2017: 4 · 2016: 1
FWCI
1.73
Type
journal article
Language
en
Open access
Licence
Locations
  • Bioengineering & Translational Medicine (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
Funders
  • National Science Foundation (DMR-1206219)
  • New York State Stem cell Science program (NYSTEM IDEA-N11G-035)
  • National Institutes of Health (R01 AR064200, AR056696, P30 ES001247)
  • University of Rochester's Health Sciences Center for Computational Innovation (HSCCI)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
  • Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
  • Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
Keywords
siRNA delivery, nanoparticle delivery, mesenchymal stem cells, diblock copolymer nanoparticles, cytotoxicity, apoptosis, innate immune activation, Toll-like receptor signaling, RIG-I-like receptor signaling, NF-κB signaling, transcriptomic changes, seeding rate, RNA interference

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