Cationic Bottlebrush Polymers Outperform Linear Polycation Analogues for pDNA Delivery and Gene Expression

Rishad J. Dalal; Ramya Kumar; Monica Ohnsorg; Mary Brown; Theresa M. Reineke

doi:10.1021/acsmacrolett.1c00335

Cationic Bottlebrush Polymers Outperform Linear Polycation Analogues for pDNA Delivery and Gene Expression

Rishad J. Dalal, Ramya Kumar, Monica Ohnsorg, Mary Brown, Theresa M. Reineke

Research output: Contribution to journal › Article › peer-review

23 Scopus citations

Abstract

Cationic polymer vehicles have emerged as promising platforms for nucleic acid delivery because of their scalability, biocompatibility, and chemical versatility. Advancements in synthetic polymer chemistry allow us to precisely tune chemical functionality with various macromolecular architectures to increase the efficacy of nonviral-based gene delivery. Herein, we demonstrate the first cationic bottlebrush polymer-mediated pDNA delivery by comparing unimolecular, synthetically defined bottlebrush polymers to their linear building blocks. We successfully synthesized poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) bottlebrushes through ring-opening metathesis polymerization to afford four bottlebrush polymers with systematic increases in backbone degree of polymerization (N_bb= 13, 20, 26, and 37), while keeping the side-chain degree of polymerization constant (N_sc= 57). Physical and chemical properties were characterized, and subsequently, the toxicity and delivery efficiency of pDNA into HEK293 cells were evaluated. The bottlebrush-pDNA complex (bottleplex) with the highestN_bb, BB_37, displayed up to a 60-fold increase in %EGFP+ cells in comparison to linear macromonomer. Additionally, we observed a trend of increasing EGFP expression with increasing polymer molecular weight. Bottleplexes and polyplexes both displayed high pDNA internalization as measured via payload enumeration per cell; however, quantitative confocal analysis revealed that bottlebrushes were able to shuttle pDNA into and around the nucleus more successfully than pDNA delivered via linear analogues. Overall, a canonical cationic monomer, such as DMAEMA, synthesized in the form of cationic bottlebrush polymers proved to be far more efficient in functional pDNA delivery and expression than linear pDMAEMA. This work underscores the importance of architectural modifications and the potential of bottlebrushes to serve as effective biomacromolecule delivery vehicles.

Original language	English (US)
Pages (from-to)	886-893
Number of pages	8
Journal	ACS Macro Letters
Volume	10
DOIs	https://doi.org/10.1021/acsmacrolett.1c00335
State	Published - Jun 25 2021

Bibliographical note

Funding Information:
We thank DARPA (Contract N660011824041) for financial support of this project and the UIC Imaging Center and Guillermo Marques for their aid in confocal analysis. We also gratefully acknowledge Craig Van Bruggen and Cristiam F. Santa Chalarca for helpful discussions and Julia Holmes for help on the TOC design.

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© 2021 American Chemical Society

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@article{42d8720a1b9d4f229979bc76e8d0bb4c,

title = "Cationic Bottlebrush Polymers Outperform Linear Polycation Analogues for pDNA Delivery and Gene Expression",

abstract = "Cationic polymer vehicles have emerged as promising platforms for nucleic acid delivery because of their scalability, biocompatibility, and chemical versatility. Advancements in synthetic polymer chemistry allow us to precisely tune chemical functionality with various macromolecular architectures to increase the efficacy of nonviral-based gene delivery. Herein, we demonstrate the first cationic bottlebrush polymer-mediated pDNA delivery by comparing unimolecular, synthetically defined bottlebrush polymers to their linear building blocks. We successfully synthesized poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) bottlebrushes through ring-opening metathesis polymerization to afford four bottlebrush polymers with systematic increases in backbone degree of polymerization (Nbb= 13, 20, 26, and 37), while keeping the side-chain degree of polymerization constant (Nsc= 57). Physical and chemical properties were characterized, and subsequently, the toxicity and delivery efficiency of pDNA into HEK293 cells were evaluated. The bottlebrush-pDNA complex (bottleplex) with the highestNbb, BB_37, displayed up to a 60-fold increase in %EGFP+ cells in comparison to linear macromonomer. Additionally, we observed a trend of increasing EGFP expression with increasing polymer molecular weight. Bottleplexes and polyplexes both displayed high pDNA internalization as measured via payload enumeration per cell; however, quantitative confocal analysis revealed that bottlebrushes were able to shuttle pDNA into and around the nucleus more successfully than pDNA delivered via linear analogues. Overall, a canonical cationic monomer, such as DMAEMA, synthesized in the form of cationic bottlebrush polymers proved to be far more efficient in functional pDNA delivery and expression than linear pDMAEMA. This work underscores the importance of architectural modifications and the potential of bottlebrushes to serve as effective biomacromolecule delivery vehicles.",

author = "Dalal, {Rishad J.} and Ramya Kumar and Monica Ohnsorg and Mary Brown and Reineke, {Theresa M.}",

note = "Funding Information: We thank DARPA (Contract N660011824041) for financial support of this project and the UIC Imaging Center and Guillermo Marques for their aid in confocal analysis. We also gratefully acknowledge Craig Van Bruggen and Cristiam F. Santa Chalarca for helpful discussions and Julia Holmes for help on the TOC design. Publisher Copyright: {\textcopyright} 2021 American Chemical Society",

year = "2021",

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doi = "10.1021/acsmacrolett.1c00335",

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T1 - Cationic Bottlebrush Polymers Outperform Linear Polycation Analogues for pDNA Delivery and Gene Expression

AU - Dalal, Rishad J.

AU - Kumar, Ramya

AU - Ohnsorg, Monica

AU - Brown, Mary

AU - Reineke, Theresa M.

N1 - Funding Information: We thank DARPA (Contract N660011824041) for financial support of this project and the UIC Imaging Center and Guillermo Marques for their aid in confocal analysis. We also gratefully acknowledge Craig Van Bruggen and Cristiam F. Santa Chalarca for helpful discussions and Julia Holmes for help on the TOC design. Publisher Copyright: © 2021 American Chemical Society

PY - 2021/6/25

Y1 - 2021/6/25

N2 - Cationic polymer vehicles have emerged as promising platforms for nucleic acid delivery because of their scalability, biocompatibility, and chemical versatility. Advancements in synthetic polymer chemistry allow us to precisely tune chemical functionality with various macromolecular architectures to increase the efficacy of nonviral-based gene delivery. Herein, we demonstrate the first cationic bottlebrush polymer-mediated pDNA delivery by comparing unimolecular, synthetically defined bottlebrush polymers to their linear building blocks. We successfully synthesized poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) bottlebrushes through ring-opening metathesis polymerization to afford four bottlebrush polymers with systematic increases in backbone degree of polymerization (Nbb= 13, 20, 26, and 37), while keeping the side-chain degree of polymerization constant (Nsc= 57). Physical and chemical properties were characterized, and subsequently, the toxicity and delivery efficiency of pDNA into HEK293 cells were evaluated. The bottlebrush-pDNA complex (bottleplex) with the highestNbb, BB_37, displayed up to a 60-fold increase in %EGFP+ cells in comparison to linear macromonomer. Additionally, we observed a trend of increasing EGFP expression with increasing polymer molecular weight. Bottleplexes and polyplexes both displayed high pDNA internalization as measured via payload enumeration per cell; however, quantitative confocal analysis revealed that bottlebrushes were able to shuttle pDNA into and around the nucleus more successfully than pDNA delivered via linear analogues. Overall, a canonical cationic monomer, such as DMAEMA, synthesized in the form of cationic bottlebrush polymers proved to be far more efficient in functional pDNA delivery and expression than linear pDMAEMA. This work underscores the importance of architectural modifications and the potential of bottlebrushes to serve as effective biomacromolecule delivery vehicles.

AB - Cationic polymer vehicles have emerged as promising platforms for nucleic acid delivery because of their scalability, biocompatibility, and chemical versatility. Advancements in synthetic polymer chemistry allow us to precisely tune chemical functionality with various macromolecular architectures to increase the efficacy of nonviral-based gene delivery. Herein, we demonstrate the first cationic bottlebrush polymer-mediated pDNA delivery by comparing unimolecular, synthetically defined bottlebrush polymers to their linear building blocks. We successfully synthesized poly(2-(dimethylamino)ethyl methacrylate) (pDMAEMA) bottlebrushes through ring-opening metathesis polymerization to afford four bottlebrush polymers with systematic increases in backbone degree of polymerization (Nbb= 13, 20, 26, and 37), while keeping the side-chain degree of polymerization constant (Nsc= 57). Physical and chemical properties were characterized, and subsequently, the toxicity and delivery efficiency of pDNA into HEK293 cells were evaluated. The bottlebrush-pDNA complex (bottleplex) with the highestNbb, BB_37, displayed up to a 60-fold increase in %EGFP+ cells in comparison to linear macromonomer. Additionally, we observed a trend of increasing EGFP expression with increasing polymer molecular weight. Bottleplexes and polyplexes both displayed high pDNA internalization as measured via payload enumeration per cell; however, quantitative confocal analysis revealed that bottlebrushes were able to shuttle pDNA into and around the nucleus more successfully than pDNA delivered via linear analogues. Overall, a canonical cationic monomer, such as DMAEMA, synthesized in the form of cationic bottlebrush polymers proved to be far more efficient in functional pDNA delivery and expression than linear pDMAEMA. This work underscores the importance of architectural modifications and the potential of bottlebrushes to serve as effective biomacromolecule delivery vehicles.

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M3 - Article

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SN - 2161-1653

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JO - ACS Macro Letters

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ER -

Cationic Bottlebrush Polymers Outperform Linear Polycation Analogues for pDNA Delivery and Gene Expression

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Cationic Bottlebrush Polymers Outperform Linear Polycation Analogues for pDNA Delivery and Gene Expression

Abstract

Bibliographical note

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University Imaging Centers

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