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Article ; Online: Technologies for controlled, local delivery of siRNA.

Sarett, Samantha M / Nelson, Christopher E / Duvall, Craig L

Journal of controlled release : official journal of the Controlled Release Society

2015 Volume 218, Page(s) 94–113

Abstract: The discovery of RNAi in the late 1990s unlocked a new realm of therapeutic possibilities by enabling potent and specific silencing of theoretically any desired genetic target. Better elucidation of the mechanism of action, the impact of chemical ... ...

Abstract	The discovery of RNAi in the late 1990s unlocked a new realm of therapeutic possibilities by enabling potent and specific silencing of theoretically any desired genetic target. Better elucidation of the mechanism of action, the impact of chemical modifications that stabilize and reduce nonspecific effects of siRNA molecules, and the key design considerations for effective delivery systems has spurred progress toward developing clinically-successful siRNA therapies. A logical aim for initial siRNA translation is local therapies, as delivering siRNA directly to its site of action helps to ensure that a sufficient dose reaches the target tissue, lessens the potential for off-target side effects, and circumvents the substantial systemic delivery barriers. While locally injected or topically applied siRNA has progressed into numerous clinical trials, an enormous opportunity exists to develop sustained-release, local delivery systems that enable both spatial and temporal control of gene silencing. This review focuses on material platforms that establish both localized and controlled gene silencing, with emphasis on the systems that show most promise for clinical translation.
MeSH term(s)	Gene Silencing ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/therapeutic use ; Technology, Pharmaceutical
Chemical Substances	RNA, Small Interfering
Language	English
Publishing date	2015-11-28
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S. ; Review
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2015.09.066
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Article ; Online: Hydrogel microspheres for spatiotemporally controlled delivery of RNA and silencing gene expression within scaffold-free tissue engineered constructs.

McMillan, Alexandra / Nguyen, Minh Khanh / Huynh, Cong Truc / Sarett, Samantha M / Ge, Peilin / Chetverikova, Melanie / Nguyen, Kien / Grosh, David / Duvall, Craig L / Alsberg, Eben

Acta biomaterialia

2021 Volume 124, Page(s) 315–326

Abstract: Delivery systems for controlled release of RNA interference (RNAi) molecules, including small interfering (siRNA) and microRNA (miRNA), have the potential to direct stem cell differentiation for regenerative musculoskeletal applications. To date, ... ...

Abstract	Delivery systems for controlled release of RNA interference (RNAi) molecules, including small interfering (siRNA) and microRNA (miRNA), have the potential to direct stem cell differentiation for regenerative musculoskeletal applications. To date, localized RNA delivery platforms in this area have focused predominantly on bulk scaffold-based approaches, which can interfere with cell-cell interactions important for recapitulating some native musculoskeletal developmental and healing processes in tissue regeneration strategies. In contrast, scaffold-free, high density human mesenchymal stem cell (hMSC) aggregates may provide an avenue for creating a more biomimetic microenvironment. Here, photocrosslinkable dextran microspheres (MS) encapsulating siRNA-micelles were prepared via an aqueous emulsion method and incorporated within hMSC aggregates for localized and sustained delivery of bioactive siRNA. siRNA-micelles released from MS in a sustained fashion over the course of 28 days, and the released siRNA retained its ability to transfect cells for gene silencing. Incorporation of fluorescently labeled siRNA (siGLO)-laden MS within hMSC aggregates exhibited tunable siGLO delivery and uptake by stem cells. Incorporation of MS loaded with siRNA targeting green fluorescent protein (siGFP) within GFP-hMSC aggregates provided sustained presentation of siGFP within the constructs and prolonged GFP silencing for up to 15 days. This platform system enables sustained gene silencing within stem cell aggregates and thus shows great potential in tissue regeneration applications. STATEMENT OF SIGNIFICANCE: This work presents a new strategy to deliver RNA-nanocomplexes from photocrosslinked dextran microspheres for tunable presentation of bioactive RNA. These microspheres were embedded within scaffold-free, human mesenchymal stem cell (hMSC) aggregates for sustained gene silencing within three-dimensional cell constructs while maintaining cell viability. Unlike exogenous delivery of RNA within culture medium that suffers from diffusion limitations and potential need for repeated transfections, this strategy provides local and sustained RNA presentation from the microspheres to cells in the constructs. This system has the potential to inhibit translation of hMSC differentiation antagonists and drive hMSC differentiation toward desired specific lineages, and is an important step in the engineering of high-density stem cell systems with incorporated instructive genetic cues for application in tissue regeneration.
MeSH term(s)	Cell Differentiation ; Gene Expression ; Gene Silencing ; Humans ; Hydrogels ; Mesenchymal Stem Cells ; Microspheres ; RNA, Small Interfering/genetics
Chemical Substances	Hydrogels ; RNA, Small Interfering
Language	English
Publishing date	2021-01-16
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2173841-5
ISSN	1878-7568 ; 1742-7061
ISSN (online)	1878-7568
ISSN	1742-7061
DOI	10.1016/j.actbio.2021.01.013
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Article ; Online: Conjugation of palmitic acid improves potency and longevity of siRNA delivered via endosomolytic polymer nanoparticles.

Sarett, Samantha M / Kilchrist, Kameron V / Miteva, Martina / Duvall, Craig L

Journal of biomedical materials research. Part A

2015 Volume 103, Issue 9, Page(s) 3107–3116

Abstract: Clinical translation of siRNA therapeutics has been limited by the inability to effectively overcome the rigorous delivery barriers associated with intracellular-acting biologics. Here, to address both potency and longevity of siRNA gene silencing, pH- ... ...

Abstract	Clinical translation of siRNA therapeutics has been limited by the inability to effectively overcome the rigorous delivery barriers associated with intracellular-acting biologics. Here, to address both potency and longevity of siRNA gene silencing, pH-responsive micellar nanoparticle (NP) carriers loaded with siRNA conjugated to palmitic acid (siRNA-PA) were investigated as a combined approach to improve siRNA endosomal escape and stability. Conjugation to hydrophobic PA improved NP loading efficiency relative to unmodified siRNA, enabling complete packaging of siRNA-PA at a lower polymer:siRNA ratio. PA conjugation also increased intracellular uptake of the nucleic acid cargo by 35-fold and produced a 3.1-fold increase in intracellular half-life. The higher uptake and improved retention of siRNA-PA NPs correlated to a 2- and 11-fold decrease in gene silencing IC50 in comparison to siRNA NPs in fibroblasts and mesenchymal stem cells, respectively, for both the model gene luciferase and the therapeutically relevant gene prolyl hydroxylase domain protein 2 (PHD2) . PA conjugation also significantly increased longevity of silencing activity following a single treatment in fibroblasts. Thus, conjugation of PA to siRNA paired with endosomolytic NPs is a promising approach to enhance the functional efficacy of siRNA in tissue regenerative and other applications.
MeSH term(s)	Animals ; Biocompatible Materials/chemistry ; Biological Transport, Active ; Drug Delivery Systems ; Endosomes/metabolism ; Gene Silencing ; HEK293 Cells ; Humans ; Materials Testing ; Mesenchymal Stromal Cells/metabolism ; Mice ; Mice, Inbred C57BL ; NIH 3T3 Cells ; Nanoparticles/chemistry ; Nanoparticles/ultrastructure ; Palmitic Acid/chemistry ; Polymers/chemistry ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/genetics ; RNA, Small Interfering/pharmacokinetics
Chemical Substances	Biocompatible Materials ; Polymers ; RNA, Small Interfering ; Palmitic Acid (2V16EO95H1)
Language	English
Publishing date	2015-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2099989-6
ISSN	1552-4965 ; 1549-3296 ; 0021-9304
ISSN (online)	1552-4965
ISSN	1549-3296 ; 0021-9304
DOI	10.1002/jbm.a.35413
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Article: Technologies for controlled, local delivery of siRNA

Sarett, Samantha M / Nelson, Christopher E / Duvall, Craig L

Journal of controlled release. 2015 Nov. 28, v. 218

2015

Abstract	The discovery of RNAi in the late 1990s unlocked a new realm of therapeutic possibilities by enabling potent and specific silencing of theoretically any desired genetic target. Better elucidation of the mechanism of action, the impact of chemical modifications that stabilize and reduce nonspecific effects of siRNA molecules, and the key design considerations for effective delivery systems has spurred progress toward developing clinically-successful siRNA therapies. A logical aim for initial siRNA translation is local therapies, as delivering siRNA directly to its site of action helps to ensure that a sufficient dose reaches the target tissue, lessens the potential for off-target side effects, and circumvents the substantial systemic delivery barriers. While locally injected or topically applied siRNA has progressed into numerous clinical trials, an enormous opportunity exists to develop sustained-release, local delivery systems that enable both spatial and temporal control of gene silencing. This review focuses on material platforms that establish both localized and controlled gene silencing, with emphasis on the systems that show most promise for clinical translation.
Keywords	mechanism of action ; RNA interference ; clinical trials ; adverse effects ; topical application ; small interfering RNA
Language	English
Dates of publication	2015-1128
Size	p. 94-113.
Publishing place	Elsevier B.V.
Document type	Article
Note	2019-12-06
ZDB-ID	632533-6
ISSN	1873-4995 ; 0168-3659
ISSN (online)	1873-4995
ISSN	0168-3659
DOI	10.1016/j.jconrel.2015.09.066
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Lipophilic siRNA targets albumin in situ and promotes bioavailability, tumor penetration, and carrier-free gene silencing.

Sarett, Samantha M / Werfel, Thomas A / Lee, Linus / Jackson, Meredith A / Kilchrist, Kameron V / Brantley-Sieders, Dana / Duvall, Craig L

Proceedings of the National Academy of Sciences of the United States of America

2017 Volume 114, Issue 32, Page(s) E6490–E6497

Abstract: Clinical translation of therapies based on small interfering RNA (siRNA) is hampered by siRNA's comprehensively poor pharmacokinetic properties, which necessitate molecule modifications and complex delivery strategies. We sought an alternative approach ... ...

Abstract	Clinical translation of therapies based on small interfering RNA (siRNA) is hampered by siRNA's comprehensively poor pharmacokinetic properties, which necessitate molecule modifications and complex delivery strategies. We sought an alternative approach to commonly used nanoparticle carriers by leveraging the long-lived endogenous serum protein albumin as an siRNA carrier. We synthesized siRNA conjugated to a diacyl lipid moiety (siRNA-L
Language	English
Publishing date	2017-08-08
Publishing country	United States
Document type	Journal Article
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.1621240114
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Article ; Online: Nanocarriers for dual delivery of doxorubicin and siRNA to overcome multidrug resistance.

Sarett, Samantha M / Beavers, Kelsey R / Miteva, Martina / Nelson, Christopher E / Duvall, Craig L

Nanomedicine (London, England)

2013 Volume 8, Issue 3, Page(s) 329–330

Language	English
Publishing date	2013-03
Publishing country	England
Document type	Comment ; Journal Article
ZDB-ID	2277839-1
ISSN	1748-6963 ; 1743-5889
ISSN (online)	1748-6963
ISSN	1743-5889
DOI	10.2217/nnm.13.9
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Article ; Online: Hydrophobic interactions between polymeric carrier and palmitic acid-conjugated siRNA improve PEGylated polyplex stability and enhance in vivo pharmacokinetics and tumor gene silencing.

Sarett, Samantha M / Werfel, Thomas A / Chandra, Irene / Jackson, Meredith A / Kavanaugh, Taylor E / Hattaway, Madison E / Giorgio, Todd D / Duvall, Craig L

Biomaterials

2016 Volume 97, Page(s) 122–132

Abstract: Formation of stable, long-circulating siRNA polyplexes is a significant challenge in translation of intravenously-delivered, polymeric RNAi cancer therapies. Here, we report that siRNA hydrophobization through conjugation to palmitic acid (siPA) improves ...

Abstract	Formation of stable, long-circulating siRNA polyplexes is a significant challenge in translation of intravenously-delivered, polymeric RNAi cancer therapies. Here, we report that siRNA hydrophobization through conjugation to palmitic acid (siPA) improves stability, in vivo pharmacokinetics, and tumor gene silencing of PEGylated nanopolyplexes (siPA-NPs) with balanced cationic and hydrophobic content in the core relative to the analogous polyplexes formed with unmodified siRNA, si-NPs. Hydrophobized siPA loaded into the NPs at a lower charge ratio (N(+):P(-)) relative to unmodified siRNA, and siPA-NPs had superior resistance to siRNA cargo unpackaging in comparison to si-NPs upon exposure to the competing polyanion heparin and serum. In vitro, siPA-NPs increased uptake in MDA-MB-231 breast cancer cells (100% positive cells vs. 60% positive cells) but exhibited equivalent silencing of the model gene luciferase relative to si-NPs. In vivo in a murine model, the circulation half-life of intravenously-injected siPA-NPs was double that of si-NPs, resulting in a >2-fold increase in siRNA biodistribution to orthotopic MDA-MB-231 mammary tumors. The increased circulation half-life of siPA-NPs was dependent upon the hydrophobic interactions of the siRNA and the NP core component and not just siRNA hydrophobization, as siPA did not contribute to improved circulation time relative to unmodified siRNA when delivered using polyplexes with a fully cationic core. Intravenous delivery of siPA-NPs also achieved significant silencing of the model gene luciferase in vivo (∼40% at 24 h after one treatment and ∼60% at 48 h after two treatments) in the murine MDA-MB-231 tumor model, while si-NPs only produced a significant silencing effect after two treatments. These data suggest that stabilization of PEGylated siRNA polyplexes through a combination of hydrophobic and electrostatic interactions between siRNA cargo and the polymeric carrier improves in vivo pharmacokinetics and tumor gene silencing relative to conventional formulations that are stabilized solely by electrostatic interactions.
Language	English
Publishing date	2016-08
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	603079-8
ISSN	1878-5905 ; 0142-9612
ISSN (online)	1878-5905
ISSN	0142-9612
DOI	10.1016/j.biomaterials.2016.04.017
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Article ; Online: Porcine Ischemic Wound-Healing Model for Preclinical Testing of Degradable Biomaterials.

Patil, Prarthana / Martin, John R / Sarett, Samantha M / Pollins, Alonda C / Cardwell, Nancy L / Davidson, Jeffrey M / Guelcher, Scott A / Nanney, Lillian B / Duvall, Craig L

Tissue engineering. Part C, Methods

2017 Volume 23, Issue 11, Page(s) 754–762

Abstract: Impaired wound healing that mimics chronic human skin pathologies is difficult to achieve in current animal models, hindering testing and development of new therapeutic biomaterials that promote wound healing. In this article, we describe a refinement ... ...

Abstract	Impaired wound healing that mimics chronic human skin pathologies is difficult to achieve in current animal models, hindering testing and development of new therapeutic biomaterials that promote wound healing. In this article, we describe a refinement and simplification of the porcine ischemic wound model that increases the size and number of experimental sites per animal. By comparing three flap geometries, we adopted a superior configuration (15 × 10 cm) that enabled testing of twenty 1 cm
MeSH term(s)	Animals ; Biocompatible Materials/pharmacology ; Blood Vessels/drug effects ; Disease Models, Animal ; Ischemia/pathology ; Macrophages/drug effects ; Macrophages/metabolism ; Materials Testing ; Skin/blood supply ; Surgical Flaps ; Sus scrofa ; Tissue Scaffolds/chemistry ; Wound Healing/drug effects
Chemical Substances	Biocompatible Materials
Language	English
Publishing date	2017-09-29
Publishing country	United States
Document type	Journal Article ; Research Support, U.S. Gov't, Non-P.H.S. ; Research Support, N.I.H., Extramural
ZDB-ID	2420585-0
ISSN	1937-3392 ; 1937-3384
ISSN (online)	1937-3392
ISSN	1937-3384
DOI	10.1089/ten.TEC.2017.0202
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Article: Hydrophobic interactions between polymeric carrier and palmitic acid-conjugated siRNA improve PEGylated polyplex stability and enhance in vivo pharmacokinetics and tumor gene silencing

Sarett, Samantha M / Werfel, Thomas A / Chandra, Irene / Jackson, Meredith A / Kavanaugh, Taylor E / Hattaway, Madison E / Giorgio, Todd D / Duvall, Craig L

Biomaterials. 2016 Aug., v. 97

2016

Abstract	Formation of stable, long-circulating siRNA polyplexes is a significant challenge in translation of intravenously-delivered, polymeric RNAi cancer therapies. Here, we report that siRNA hydrophobization through conjugation to palmitic acid (siPA) improves stability, in vivo pharmacokinetics, and tumor gene silencing of PEGylated nanopolyplexes (siPA-NPs) with balanced cationic and hydrophobic content in the core relative to the analogous polyplexes formed with unmodified siRNA, si-NPs. Hydrophobized siPA loaded into the NPs at a lower charge ratio (N+:P−) relative to unmodified siRNA, and siPA-NPs had superior resistance to siRNA cargo unpackaging in comparison to si-NPs upon exposure to the competing polyanion heparin and serum. In vitro, siPA-NPs increased uptake in MDA-MB-231 breast cancer cells (100% positive cells vs. 60% positive cells) but exhibited equivalent silencing of the model gene luciferase relative to si-NPs. In vivo in a murine model, the circulation half-life of intravenously-injected siPA-NPs was double that of si-NPs, resulting in a >2-fold increase in siRNA biodistribution to orthotopic MDA-MB-231 mammary tumors. The increased circulation half-life of siPA-NPs was dependent upon the hydrophobic interactions of the siRNA and the NP core component and not just siRNA hydrophobization, as siPA did not contribute to improved circulation time relative to unmodified siRNA when delivered using polyplexes with a fully cationic core. Intravenous delivery of siPA-NPs also achieved significant silencing of the model gene luciferase in vivo (∼40% at 24 h after one treatment and ∼60% at 48 h after two treatments) in the murine MDA-MB-231 tumor model, while si-NPs only produced a significant silencing effect after two treatments. These data suggest that stabilization of PEGylated siRNA polyplexes through a combination of hydrophobic and electrostatic interactions between siRNA cargo and the polymeric carrier improves in vivo pharmacokinetics and tumor gene silencing relative to conventional formulations that are stabilized solely by electrostatic interactions.
Keywords	RNA interference ; animal models ; anions ; blood serum ; breast neoplasms ; electrostatic interactions ; genes ; half life ; heparin ; hydrophobic bonding ; hydrophobicity ; intravenous injection ; luciferase ; mammary neoplasms (animal) ; mice ; neoplasm cells ; palmitic acid ; pharmacokinetics ; small interfering RNA
Language	English
Dates of publication	2016-08
Size	p. 122-132.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	603079-8
ISSN	0142-9612
ISSN	0142-9612
DOI	10.1016/j.biomaterials.2016.04.017
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Endothelial cell attachment and shear response on biomimetic polymer-coated vascular grafts.

Dudash, Lynn A / Kligman, Faina / Sarett, Samantha M / Kottke-Marchant, Kandice / Marchant, Roger E

Journal of biomedical materials research. Part A

2012 Volume 100, Issue 8, Page(s) 2204–2210

Abstract: Endothelial cell (EC) adhesion, shear retention, morphology, and hemostatic gene expression on fibronectin (FN) and RGD fluorosurfactant polymer (FSP)-coated expanded polytetrafluoroethylene grafts were investigated using an in vitro perfusion system. ... ...

Abstract	Endothelial cell (EC) adhesion, shear retention, morphology, and hemostatic gene expression on fibronectin (FN) and RGD fluorosurfactant polymer (FSP)-coated expanded polytetrafluoroethylene grafts were investigated using an in vitro perfusion system. ECs were sodded on both types of grafts and exposed to 8 dyn/cm(2) of shear stress. After 24 h, the EC retention on RGD-FSP-coated grafts was 59 ± 14%, which is statistically higher than the 36 ± 11% retention measured on FN grafts (p < 0.02). Additionally, ECs on RGD-FSP exhibited a more spread morphology and oriented in the direction of shear stress, as demonstrated by actin fiber staining. This spread morphology has been observed earlier in cells that are adapting to shear stress. Real-time PCR for vascular cell adhesion molecule 1, tissue factor, tissue plasminogen activator, and inducible nitric oxide synthase indicated that the RGD-FSP material did not activate the cells and that shear stress appears to induce a more vasoprotective phenotype, as shown by a significant decrease in VCAM-1 expression, compared with sodded grafts. RGD-FSP-coating allows for a cell layer that is more resistant to physiological shear stress, as shown by the increased cell retention over FN. This shear stable EC layer is necessary for in vivo endothelialization of the graft material, which shows promise to increase the patency of synthetic small diameter vascular grafts.
MeSH term(s)	Actin Cytoskeleton/metabolism ; Biomimetic Materials/chemistry ; Biomimetic Materials/pharmacology ; Blood Vessel Prosthesis ; Cell Adhesion/drug effects ; Cell Shape/drug effects ; Coated Materials, Biocompatible/chemistry ; Coated Materials, Biocompatible/pharmacology ; Endothelial Cells/cytology ; Endothelial Cells/drug effects ; Gene Expression Regulation/drug effects ; Humans ; Microscopy, Fluorescence ; Oligopeptides/pharmacology ; Perfusion ; Polytetrafluoroethylene/pharmacology ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Real-Time Polymerase Chain Reaction ; Stress, Mechanical ; Surface-Active Agents/pharmacology ; Vascular Cell Adhesion Molecule-1/genetics ; Vascular Cell Adhesion Molecule-1/metabolism ; Water/chemistry
Chemical Substances	Coated Materials, Biocompatible ; Oligopeptides ; RNA, Messenger ; Surface-Active Agents ; Vascular Cell Adhesion Molecule-1 ; Water (059QF0KO0R) ; arginyl-glycyl-aspartic acid (78VO7F77PN) ; Polytetrafluoroethylene (9002-84-0)
Language	English
Publishing date	2012-05-24
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	2099989-6
ISSN	1552-4965 ; 1549-3296 ; 0021-9304
ISSN (online)	1552-4965
ISSN	1549-3296 ; 0021-9304
DOI	10.1002/jbm.a.34119
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