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Article ; Online: Advances in the delivery of RNA therapeutics: from concept to clinical reality.

Kaczmarek, James C / Kowalski, Piotr S / Anderson, Daniel G

2017 Volume 9, Issue 1, Page(s) 60

Abstract: The rapid expansion of the available genomic data continues to greatly impact biomedical science and medicine. Fulfilling the clinical potential of genetic discoveries requires the development of therapeutics that can specifically modulate the expression ...

Abstract	The rapid expansion of the available genomic data continues to greatly impact biomedical science and medicine. Fulfilling the clinical potential of genetic discoveries requires the development of therapeutics that can specifically modulate the expression of disease-relevant genes. RNA-based drugs, including short interfering RNAs and antisense oligonucleotides, are particularly promising examples of this newer class of biologics. For over two decades, researchers have been trying to overcome major challenges for utilizing such RNAs in a therapeutic context, including intracellular delivery, stability, and immune response activation. This research is finally beginning to bear fruit as the first RNA drugs gain FDA approval and more advance to the final phases of clinical trials. Furthermore, the recent advent of CRISPR, an RNA-guided gene-editing technology, as well as new strides in the delivery of messenger RNA transcribed in vitro, have triggered a major expansion of the RNA-therapeutics field. In this review, we discuss the challenges for clinical translation of RNA-based therapeutics, with an emphasis on recent advances in delivery technologies, and present an overview of the applications of RNA-based drugs for modulation of gene/protein expression and genome editing that are currently being investigated both in the laboratory as well as in the clinic.
MeSH term(s)	Gene Editing ; Genetic Therapy/methods ; Humans ; Oligonucleotides, Antisense/therapeutic use ; RNA, Small Interfering/therapeutic use
Chemical Substances	Oligonucleotides, Antisense ; RNA, Small Interfering
Language	English
Publishing date	2017-06-27
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2484394-5
ISSN	1756-994X ; 1756-994X
ISSN (online)	1756-994X
ISSN	1756-994X
DOI	10.1186/s13073-017-0450-0
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Systemic delivery of mRNA and DNA to the lung using polymer-lipid nanoparticles.

Kaczmarek, James C / Patel, Asha Kumari / Rhym, Luke H / Palmiero, Umberto Capasso / Bhat, Balkrishen / Heartlein, Michael W / DeRosa, Frank / Anderson, Daniel G

Biomaterials

2021 Volume 275, Page(s) 120966

Abstract: Non-viral vectors offer the potential to deliver nucleic acids including mRNA and DNA into cells in vivo. However, designing materials that effectively deliver to target organs and then to desired compartments within the cell remains a challenge. Here we ...

Abstract	Non-viral vectors offer the potential to deliver nucleic acids including mRNA and DNA into cells in vivo. However, designing materials that effectively deliver to target organs and then to desired compartments within the cell remains a challenge. Here we develop polymeric materials that can be optimized for either DNA transcription in the nucleus or mRNA translation in the cytosol. We synthesized poly(beta amino ester) terpolymers (PBAEs) with modular changes to monomer chemistry to investigate influence on nucleic acid delivery. We identified two PBAEs with a single monomer change as being effective for either DNA (D-90-C12-103) or mRNA (DD-90-C12-103) delivery to lung endothelium following intravenous injection in mice. Physical properties such as particle size or charge did not account for the difference in transfection efficacy. However, endosome co-localization studies revealed that D-90-C12-103 nanoparticles resided in late endosomes to a greater extent than DD-90-C12-103. We compared luciferase expression in vivo and observed that, even with nucleic acid optimized vectors, peak luminescence using mRNA was two orders of magnitude greater than pDNA in the lungs of mice following systemic delivery. This study indicates that different nucleic acids require tailored delivery vectors, and further support the potential of PBAEs as intracellular delivery materials.
MeSH term(s)	Animals ; DNA ; Lipids ; Lung ; Mice ; Nanoparticles ; Polymers ; RNA, Messenger/genetics ; Transfection
Chemical Substances	Lipids ; Polymers ; RNA, Messenger ; DNA (9007-49-2)
Language	English
Publishing date	2021-06-10
Publishing country	Netherlands
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	603079-8
ISSN	1878-5905 ; 0142-9612
ISSN (online)	1878-5905
ISSN	0142-9612
DOI	10.1016/j.biomaterials.2021.120966
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Article: Systemic delivery of mRNA and DNA to the lung using polymer-lipid nanoparticles

Kaczmarek, James C. / Patel, Asha Kumari / Rhym, Luke H. / Palmiero, Umberto Capasso / Bhat, Balkrishen / Heartlein, Michael W. / DeRosa, Frank / Anderson, Daniel G.

Biomaterials. 2021 Aug., v. 275

2021

Abstract	Non-viral vectors offer the potential to deliver nucleic acids including mRNA and DNA into cells in vivo. However, designing materials that effectively deliver to target organs and then to desired compartments within the cell remains a challenge. Here we develop polymeric materials that can be optimized for either DNA transcription in the nucleus or mRNA translation in the cytosol. We synthesized poly(beta amino ester) terpolymers (PBAEs) with modular changes to monomer chemistry to investigate influence on nucleic acid delivery. We identified two PBAEs with a single monomer change as being effective for either DNA (D-90-C12-103) or mRNA (DD-90-C12-103) delivery to lung endothelium following intravenous injection in mice. Physical properties such as particle size or charge did not account for the difference in transfection efficacy. However, endosome co-localization studies revealed that D-90-C12-103 nanoparticles resided in late endosomes to a greater extent than DD-90-C12-103. We compared luciferase expression in vivo and observed that, even with nucleic acid optimized vectors, peak luminescence using mRNA was two orders of magnitude greater than pDNA in the lungs of mice following systemic delivery. This study indicates that different nucleic acids require tailored delivery vectors, and further support the potential of PBAEs as intracellular delivery materials.
Keywords	DNA ; biocompatible materials ; chemistry ; cytosol ; endosomes ; endothelium ; intravenous injection ; luciferase ; luminescence ; lungs ; particle size ; polymers ; transfection
Language	English
Dates of publication	2021-08
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	603079-8
ISSN	0142-9612
ISSN	0142-9612
DOI	10.1016/j.biomaterials.2021.120966
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Article ; Online: The role of emotions in esports performance.

Behnke, Maciej / Gross, James J / Kaczmarek, Lukasz D

Emotion (Washington, D.C.)

2020 Volume 22, Issue 5, Page(s) 1059–1070

Abstract	Emotions that differ on the approach-avoidance dimension are thought to have different functions. Based on the motivational dimensional model of affect, we expected high-approach tendency (and not valence) to facilitate sports performance in a gaming context. Moreover, we expected the influence of high-approach emotions on performance to be mediated by higher levels of cognitive and physiological challenge as an approach-related response. To test these hypotheses, 241 men completed 5 matches of a soccer video game FIFA 19. Before each match, approach tendencies and valence were experimentally manipulated by showing films that elicit amusement, enthusiasm, sadness, anger, and neutral states. Approach tendency, challenge/threat evaluations, cardiovascular responses, and game scores were recorded. After watching enthusiastic and amusing videos, gamers displayed stronger approach tendencies, and, in turn, improved performance, compared to negative emotions and neutral conditions. Moreover, enthusiasm produced a stronger approach tendency and promoted better performance than amusement. Elicitation of unpleasant emotions (anger and sadness) had no effect on approach tendencies or gaming-outcomes relative to the neutral conditions. Across all conditions, gamers with higher levels of cognitive and cardiovascular challenge achieved higher scores. These findings indicate that in a gaming context performance is enhanced by pleasant emotions with high-approach tendencies. (PsycInfo Database Record (c) 2022 APA, all rights reserved).
MeSH term(s)	Anger/physiology ; Emotions/physiology ; Humans ; Male ; Motivation ; Sadness/psychology ; Video Games/psychology
Language	English
Publishing date	2020-10-29
Publishing country	United States
Document type	Journal Article
ZDB-ID	2102391-8
ISSN	1931-1516 ; 1528-3542
ISSN (online)	1931-1516
ISSN	1528-3542
DOI	10.1037/emo0000903
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Article ; Online: Poly(β-amino ester)-co-poly(caprolactone) Terpolymers as Nonviral Vectors for mRNA Delivery In Vitro and In Vivo.

Capasso Palmiero, Umberto / Kaczmarek, James C / Fenton, Owen S / Anderson, Daniel G

Advanced healthcare materials

2018 Volume 7, Issue 14, Page(s) e1800249

Abstract: The production of new proteins with messenger RNA (mRNA) has gained a broad interest due to its potential for addressing a wide range of diseases. Here, the design and characterization of novel ionizable poly(β-amino ester)-co-poly(caprolactone) ... ...

Abstract	The production of new proteins with messenger RNA (mRNA) has gained a broad interest due to its potential for addressing a wide range of diseases. Here, the design and characterization of novel ionizable poly(β-amino ester)-co-poly(caprolactone) terpolymers, synthesized via the combination of the ring opening polymerization and the Michael step-growth polymerization, are reported. The versatility of this method is demonstrated by varying the number of caprolactone units attached to each poly(β-amino ester) (PBAE) terpolymer. The ability of the novel poly-caprolactone (PCL)-based PBAE materials to deliver mRNA is shown to depend on the physiochemical characteristics of the material, such as lipophilicity, as well as the formulation method used to complex the polymer with the oligonucleotide. This latter variable represents a previously unstudied aspect of PBAE library screens that can play an important role in identifying true top candidates for nucleic acid delivery. The most stable terpolymer is injected intravenously (IV) in mice and shows a transfection efficacy several times higher than the polyethylenimine (PEI) which is focused in the spleen, opening the possibility to use these biodegradable carriers in the intravenous delivery of antigen-encoding mRNA for cancer immunotherapy and vaccination.
MeSH term(s)	Cell Survival/physiology ; Genetic Vectors/chemistry ; HeLa Cells ; Humans ; Nanoparticles/chemistry ; Polyesters/chemistry ; Polyethyleneimine/chemistry ; Polymers/chemistry ; RNA, Messenger/chemistry
Chemical Substances	Polyesters ; Polymers ; RNA, Messenger ; poly(beta-amino ester) ; polycaprolactone (24980-41-4) ; Polyethyleneimine (9002-98-6)
Language	English
Publishing date	2018-05-14
Publishing country	Germany
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	2649576-4
ISSN	2192-2659 ; 2192-2640
ISSN (online)	2192-2659
ISSN	2192-2640
DOI	10.1002/adhm.201800249
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Article: Extension of the 5-alkynyluridine side chain via C-C-bond formation in modified organometallic nucleosides using the Nicholas reaction.

Kaczmarek, Renata / Korczyński, Dariusz / Green, James R / Dembinski, Roman

Beilstein journal of organic chemistry

2020 Volume 16, Page(s) 1–8

Abstract: ... react with diverse C-nucleophiles. Synthetic outcomes confirmed that the Nicholas reaction can be ...

Abstract	Dicobalt hexacarbonyl nucleoside complexes of propargyl ether or esters of 5-substituted uridines react with diverse C-nucleophiles. Synthetic outcomes confirmed that the Nicholas reaction can be carried out in a nucleoside presence, leading to a divergent synthesis of novel metallo-nucleosides enriched with alkene, arene, arylketo, and heterocyclic functions, in the deoxy and ribo series.
Language	English
Publishing date	2020-01-02
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2192461-2
ISSN	1860-5397
ISSN	1860-5397
DOI	10.3762/bjoc.16.1
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Article ; Online: Advances in the delivery of RNA therapeutics

James C. Kaczmarek / Piotr S. Kowalski / Daniel G. Anderson

Genome Medicine, Vol 9, Iss 1, Pp 1-

from concept to clinical reality

2017 Volume 16

Abstract: Abstract The rapid expansion of the available genomic data continues to greatly impact biomedical science and medicine. Fulfilling the clinical potential of genetic discoveries requires the development of therapeutics that can specifically modulate the ... ...

Abstract	Abstract The rapid expansion of the available genomic data continues to greatly impact biomedical science and medicine. Fulfilling the clinical potential of genetic discoveries requires the development of therapeutics that can specifically modulate the expression of disease-relevant genes. RNA-based drugs, including short interfering RNAs and antisense oligonucleotides, are particularly promising examples of this newer class of biologics. For over two decades, researchers have been trying to overcome major challenges for utilizing such RNAs in a therapeutic context, including intracellular delivery, stability, and immune response activation. This research is finally beginning to bear fruit as the first RNA drugs gain FDA approval and more advance to the final phases of clinical trials. Furthermore, the recent advent of CRISPR, an RNA-guided gene-editing technology, as well as new strides in the delivery of messenger RNA transcribed in vitro, have triggered a major expansion of the RNA-therapeutics field. In this review, we discuss the challenges for clinical translation of RNA-based therapeutics, with an emphasis on recent advances in delivery technologies, and present an overview of the applications of RNA-based drugs for modulation of gene/protein expression and genome editing that are currently being investigated both in the laboratory as well as in the clinic.
Keywords	Antisense oligonucleotide ; Clinical trial ; CRISPR ; Gene editing ; Gene therapy ; Messenger RNA delivery ; Medicine ; R ; Genetics ; QH426-470
Subject code	610
Language	English
Publishing date	2017-06-01T00:00:00Z
Publisher	BMC
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Highly Tunable Thiol-Ene Photoresins for Volumetric Additive Manufacturing.

Cook, Caitlyn C / Fong, Erika J / Schwartz, Johanna J / Porcincula, Dominique H / Kaczmarek, Allison C / Oakdale, James S / Moran, Bryan D / Champley, Kyle M / Rackson, Charles M / Muralidharan, Archish / McLeod, Robert R / Shusteff, Maxim

Advanced materials (Deerfield Beach, Fla.)

2020 Volume 32, Issue 47, Page(s) e2003376

Abstract: Volumetric additive manufacturing (VAM) forms complete 3D objects in a single photocuring operation without layering defects, enabling 3D printed polymer parts with mechanical properties similar to their bulk material counterparts. This study presents ... ...

Abstract	Volumetric additive manufacturing (VAM) forms complete 3D objects in a single photocuring operation without layering defects, enabling 3D printed polymer parts with mechanical properties similar to their bulk material counterparts. This study presents the first report of VAM-printed thiol-ene resins. With well-ordered molecular networks, thiol-ene chemistry accesses polymer materials with a wide range of mechanical properties, moving VAM beyond the limitations of commonly used acrylate formulations. Since free-radical thiol-ene polymerization is not inhibited by oxygen, the nonlinear threshold response required in VAM is introduced by incorporating 2,2,6,6-tetramethyl-1-piperidinyloxy (TEMPO) as a radical scavenger. Tuning of the reaction kinetics is accomplished by balancing inhibitor and initiator content. Coupling this with quantitative measurements of the absorbed volumetric optical dose allows control of polymer conversion and gelation during printing. Importantly, this work thereby establishes the first comprehensive framework for spatial-temporal control over volumetric energy distribution, demonstrating structures 3D printed in thiol-ene resin by means of tomographic volumetric VAM. Mechanical characterization of this thiol-ene system, with varied ratios of isocyanurate and triethylene glycol monomers, reveals highly tunable mechanical response far more versatile than identical acrylate-based resins. This broadens the range of materials and properties available for VAM, taking another step toward high-performance printed polymers.
Language	English
Publishing date	2020-10-01
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1474949-X
ISSN	1521-4095 ; 0935-9648
ISSN (online)	1521-4095
ISSN	0935-9648
DOI	10.1002/adma.202003376
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Article ; Online: Optimization of a Degradable Polymer-Lipid Nanoparticle for Potent Systemic Delivery of mRNA to the Lung Endothelium and Immune Cells.

Kaczmarek, James C / Kauffman, Kevin J / Fenton, Owen S / Sadtler, Kaitlyn / Patel, Asha K / Heartlein, Michael W / DeRosa, Frank / Anderson, Daniel G

Nano letters

2018 Volume 18, Issue 10, Page(s) 6449–6454

Abstract: mRNA therapeutics hold great potential for treating a variety of diseases through protein-replacement, immunomodulation, and gene editing. However, much like siRNA therapy the majority of progress in mRNA delivery has been confined to the liver. ... ...

Abstract	mRNA therapeutics hold great potential for treating a variety of diseases through protein-replacement, immunomodulation, and gene editing. However, much like siRNA therapy the majority of progress in mRNA delivery has been confined to the liver. Previously, we demonstrated that poly(β-amino esters), a class of degradable polymers, are capable of systemic mRNA delivery to the lungs in mice when formulated into nanoparticles with poly(ethylene glycol)-lipid conjugates. Using experimental design, a statistical approach to optimization that reduces experimental burden, we demonstrate herein that these degradable polymer-lipid nanoparticles can be optimized in terms of polymer synthesis and nanoparticle formulation to achieve a multiple order-of-magnitude increase in potency. Furthermore, using genetically engineered Cre reporter mice, we demonstrate that mRNA is functionally delivered to both the lung endothelium and pulmonary immune cells, expanding the potential utility of these nanoparticles.
MeSH term(s)	Animals ; Endothelium/drug effects ; Endothelium/immunology ; Endothelium/pathology ; Gene Transfer Techniques ; Humans ; Lipids/administration & dosage ; Lipids/chemistry ; Lung/drug effects ; Lung/immunology ; Lung/pathology ; Mice ; Nanoparticles/administration & dosage ; Nanoparticles/chemistry ; Polyethylene Glycols/chemistry ; Polymers/chemistry ; RNA, Messenger/chemistry ; RNA, Messenger/genetics ; RNA, Small Interfering/administration & dosage ; RNA, Small Interfering/chemistry ; RNA, Small Interfering/genetics
Chemical Substances	Lipids ; Polymers ; RNA, Messenger ; RNA, Small Interfering ; Polyethylene Glycols (3WJQ0SDW1A)
Language	English
Publishing date	2018-09-20
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1530-6992
ISSN (online)	1530-6992
DOI	10.1021/acs.nanolett.8b02917
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Article ; Online: Adjusting biomaterial composition to achieve controlled multiple-day release of dexamethasone from an extended-wear silicone hydrogel contact lens.

Kaczmarek, James C / Tieppo, Arianna / White, Charles J / Byrne, Mark E

Journal of biomaterials science. Polymer edition

2014 Volume 25, Issue 1, Page(s) 88–100

Abstract: Purpose: To alter the composition and structure of silicone hydrogel contact lenses to achieve controlled release of dexamethasone and evaluate the lens optical and mechanical properties compared to commercial lenses. There is a tremendous need for ... ...

Abstract	Purpose: To alter the composition and structure of silicone hydrogel contact lenses to achieve controlled release of dexamethasone and evaluate the lens optical and mechanical properties compared to commercial lenses. There is a tremendous need for controlled release of drugs from ocular biomaterials as the majority of ophthalmic drugs are delivered via topical eye drops, which have low bioavailability and patient compliance. Methods: Poly(PDMS-co-TRIS-co-DMA) contact lenses were synthesized with varying PDMS/TRIS:DMA ratios (0.25:1, 0.67:1, 1.5:1) as well as with additional crosslinking monomers. Lenses were characterized via in vitro release studies in a microfluidic device at ocular flowrates and in large well-mixed volumes, optical quality studies over visible wavelengths, mechanical analysis, and determination of polymer volume fraction in the swollen state. Results: Extended and controlled release of therapeutically relevant concentrations of dexamethasone was achieved for multiple day, continuous wear up to 60 days at in vitro ocular flowrates. Release was delayed due to a combination of increased hydrophobic to hydrophilic composition and the inclusion of additional structural constraints, both of which decreased the polymer volume fraction in the swollen state. However, decreased mass release rates were at the expense of increased modulus and decreased lens flexibility. All lenses had high optical clarity (∼90% transmittance) and contained highly oxygen permeable siloxane composition similar to those found in commercial silicone hydrogel lenses, but they had poor flexibility for use as soft contact lenses. Conclusions: Based on our results, the lenses described herein likely have too high of a modulus for use as extended-wear, soft contact lenses with drug release. Therefore, other controlled release methods would be better suited for maintaining adequate mechanical properties and achieving controlled and extended release for the duration of wear in soft, silicone hydrogel contact lens biomaterials. However, these biomaterials may find clinical use as more rigid gas permeable contact lenses or inserts.
MeSH term(s)	Biocompatible Materials/chemistry ; Contact Lenses, Extended-Wear ; Delayed-Action Preparations ; Dexamethasone/administration & dosage ; Dexamethasone/chemistry ; Drug Carriers/chemistry ; Hydrogels/chemistry ; Silicones/chemistry
Chemical Substances	Biocompatible Materials ; Delayed-Action Preparations ; Drug Carriers ; Hydrogels ; Silicones ; Dexamethasone (7S5I7G3JQL)
Language	English
Publishing date	2014
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	1014190-x
ISSN	1568-5624 ; 1568-5616 ; 0920-5063
ISSN (online)	1568-5624 ; 1568-5616
ISSN	0920-5063
DOI	10.1080/09205063.2013.840228
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