Poly(2-deoxy-2-methacrylamido glucopyranose)-<i>b</i>-Poly(methacrylate amine)s: Optimization of Diblock Glycopolycations for Nucleic Acid Delivery

Li, Haibo; Cortez, Mallory A.; Phillips, Haley; Wu, Yaoying; Reineke, Theresa M.

doi:10.1021/mz300660t

Cited by 44 publications

(77 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…9,10 The polyplexes formed from this series of diblock glycopolymers exhibited colloidal stability in cell media and had relatively low cytotoxicity. Moreover, both amine type and cationic charge block length were demonstrated to greatly affect transgene expression efficiency in vitro.…”

Section: Resultsmentioning

confidence: 98%

Glucose-Containing Diblock Polycations Exhibit Molecular Weight, Charge, and Cell-Type Dependence for pDNA Delivery

Wang

Sprouse

et al. 2014

Biomacromolecules

Self Cite

View full text Add to dashboard Cite

A series of diblock glycopolycations were created by polymerizing 2-deoxy-2-methacrylamido glucopyranose (MAG) with either a tertiary amine-containing monomer, N-[3-(N,N-dimethylamino) propyl] methacrylamide (DMAPMA), or a primary amine-containing unit, N-(2-aminoethyl) methacrylamide (AEMA). Seven structures were synthesized via aqueous reversible addition–fragmentation chain transfer (RAFT) polymerization that varied in the block lengths of MAG, DMAPMA, and AEMA along with two homopolymer controls of DMAPMA and AEMA that lacked a MAG block. The polymers were all able to complex plasmid DNA into polyplex structures and to prevent colloidal aggregation of polyplexes in physiological salt conditions. In vitro transfection experiments were performed in both HeLa (human cervix adenocarcinoma) cells and HepG2 (human liver hepatocellular carcinoma) cells to examine the role of charge type, block length, and cell type on transfection efficiency and toxicity. The glycopolycation vehicles with primary amine blocks and PAEMA homopolymers revealed much higher transfection efficiency and lower toxicity when compared to analogs created with DMAPMA. Block length was also shown to influence cellular delivery and toxicity; as the block length of DMAPMA increased in the glycopolycation-based polyplexes, toxicity increased while transfection decreased. While the charge block played a major role in delivery, the MAG block length did not affect these cellular parameters. Lastly, cell type played a major role in efficiency. These glycopolymers revealed higher cellular uptake and transfection efficiency in HepG2 cells than in HeLa cells, while homopolycations (PAEMA and PDMAPMA) lacking the MAG blocks exhibited the opposite trend, signifying that the MAG block could aid in hepatocyte transfection.

show abstract

Section: Resultsmentioning

confidence: 98%

Glucose-Containing Diblock Polycations Exhibit Molecular Weight, Charge, and Cell-Type Dependence for pDNA Delivery

Wang

Sprouse

et al. 2014

Biomacromolecules

Self Cite

View full text Add to dashboard Cite

show abstract

“…Also, it should be noted that both pMAT- b -AEMA-2 and- 3 polyplex formulations at N/P = 7 had cell uptake comparable to that of PEG- b -AEMA. 23,44 …”

Section: Resultsmentioning

confidence: 99%

“…It should be noted that differences between some samples are not statistically significant, but the general trend that appears in the uptake assays as well as gene expression studies for both cells lines correlates with previous work in the field. 23,41,44,46 …”

Section: Resultsmentioning

confidence: 99%

Trehalose-Based Block Copolycations Promote Polyplex Stabilization for Lyophilization and in Vivo pDNA Delivery

Tolstyka

Phillips

Cortez

et al. 2015

ACS Biomater. Sci. Eng.

Self Cite

View full text Add to dashboard Cite

The development and thorough characterization of nonviral delivery agents for nucleic acid and genome editing therapies are of high interest to the field of nanomedicine. Indeed, this vehicle class offers the ability to tune chemical architecture/biological activity and readily package nucleic acids of various sizes and morphologies for a variety of applications. Herein, we present the synthesis and characterization of a class of trehalose-based block copolycations designed to stabilize polyplex formulations for lyophilization and in vivo administration. A 6-methacrylamido-6-deoxy trehalose (MAT) monomer was synthesized from trehalose and polymerized via reversible addition–fragmentation chain transfer (RAFT) polymerization to yield pMAT43. The pMAT43 macro-chain transfer agent was then chain-extended with aminoethylmethacrylamide (AEMA) to yield three different pMAT-b-AEMA cationic-block copolymers, pMAT-b-AEMA-1 (21 AEMA repeats), -2 (44 AEMA repeats), and -3 (57 AEMA repeats). These polymers along with a series of controls were used to form polyplexes with plasmids encoding firefly luciferase behind a strong ubiquitous promoter. The trehalose-coated polyplexes were characterized in detail and found to be resistant to colloidal aggregation in culture media containing salt and serum. The trehalose-polyplexes also retained colloidal stability and promoted high gene expression following lyophilization and reconstitution. Cytotoxicity, cellular uptake, and transfection ability were assessed in vitro using both human glioblastoma (U87) and human liver carcinoma (HepG2) cell lines wherein pMAT-b-AEMA-2 was found to have the optimal combination of high gene expression and low toxicity. pMAT-b-AEMA-2 polyplexes were evaluated in mice via slow tail vein infusion. The vehicle displayed minimal toxicity and discouraged nonspecific internalization in the liver, kidney, spleen, and lungs as determined by quantitative polymerase chain reaction (qPCR) and fluorescence imaging experiments. Hydrodynamic infusion of the polyplexes, however, led to very specific localization of the polyplexes to the mouse liver and promoted excellent gene expression in vivo.

show abstract

“…The diblock glycopolymers however, gave a low zeta potential value when it was dissolved in acetate buffer as polymer solutions were prepared at 1 mg mL −1 of polymer of acetate buffer solution, therefore resulting in less cationic groups in the same volume of solutions. It has also been reported that excessive positive charges can lead to cytotoxicity and therefore the carbohydrate groups here can assist by lowing the toxicity and preventing colloidal aggregation in cell culture media …”

Section: Resultsmentioning

confidence: 94%

“…Trehalose which comprise of two linked (1→1) α ‐glucose units increased the stability of formed polyplexes for lyophilization and the delivery of pDNA in vivo . In the same research group, poly(2‐deoxy‐2‐methacrylamido glucopyranose)‐ b ‐poly(methacrylate amine) were synthesized and found that the glucose based cationic glycopolymers resulted in very effective pDNA delivery and polymers displayed low cytotoxicity profiles . However, there are only a few articles reporting the synthesis of mannose based cationic glycopolymers .…”

Section: Introductionmentioning

confidence: 99%

Acetyl‐α‐d‐mannopyranose‐based cationic polymer via RAFT polymerization for lectin and nucleic acid bindings

Ting

Min

Lau

et al. 2017

J of Applied Polymer Sci

View full text Add to dashboard Cite

Functional cationic polymers carrying mannose moieties were synthesized in a facile manner by employing RAFT polymerization. Initially, a protected carbohydrate based monomer, [2-(2,3,4,6-tetra-O-acetyl-a-D-mannopyranosyloxy)ethyl methacrylate (AcManEMA)], was prepared by the O-glycosylation of 2-hydroxyethyl methacrylate (HEMA). Subsequently, a macroRAFT agent of poly[2-(dimethyl)amino ethyl methacrylate] (PDMAEMA) was generated, and a further chain extension polymerization with AcMa-nEMA was carried out in dioxane to form a acetylated mannose cationic diblock copolymer, PDMAEMA-b-PAcManEMA. It was attained in high yields and displayed low dispersity (Ð). Acetylated mannose moieties on the polymer were deprotected with sodium methoxide and the amines from the DMAEMA block were protonated to yield a cationic diblock glycopolymer, PDMAEMA-b-PMa-nEMA. The cationic property of polymers were characterized by mixing with a negatively charged siRNA duplex and a pDNA, and aggregates of 102 and 233 nm were obtained, respectively. Agarose gel shift assay revealed that the polymers were able to retain the nucleic acids as large polymer complexes. Lectin binding assay proved that the mannose residue on the polymers were only able to bind specifically with ConA. PNA lectin was employed as a control and did not show specific binding. The cationic glycopolymer could be advantageous in targeted nucleic acids delivery in specific cells.

show abstract

Poly(2-deoxy-2-methacrylamido glucopyranose)-b-Poly(methacrylate amine)s: Optimization of Diblock Glycopolycations for Nucleic Acid Delivery

Cited by 44 publications

References 28 publications

Glucose-Containing Diblock Polycations Exhibit Molecular Weight, Charge, and Cell-Type Dependence for pDNA Delivery

Glucose-Containing Diblock Polycations Exhibit Molecular Weight, Charge, and Cell-Type Dependence for pDNA Delivery

Trehalose-Based Block Copolycations Promote Polyplex Stabilization for Lyophilization and in Vivo pDNA Delivery

Acetyl‐α‐d‐mannopyranose‐based cationic polymer via RAFT polymerization for lectin and nucleic acid bindings

Contact Info

Product

Resources

About