Abstract: The wealth of information being obtained from genomic, proteomic, and glycomic research is allowing researchers to unravel the intricate genetic and epigenetic mechanisms associated with human health and disease. The intracellular delivery of nucleic acids to study these processes offers unprecedented promise for revolutionizing biomedical research and drug development. However, the nucleic acid delivery vehicle plays a central yet elusive role in dictating the efficacy, safety, mechanisms, and kinetics of gene regulation in a spatial and temporal manner; thus, having a far-reaching impact in health-related research. To this end, we have developed several novel carbohydrate-containing polymers that have shown outstanding affinity to encapsulate polynucleotides into nanoparticles (polyplexes) and facilitate highly efficient intracellular delivery without toxicity. The goals of this project directly commence from our previous work where we aim to examine our wide-range of delivery vehicles for their mechanistic pathways and kinetics of nucleic acid encapsulation and intracellular transport from the cell surface to their final intracellular destination. We plan to examine 10 different carbohydrate-based polymers synthesized in our laboratory for their delivery mechanisms and kinetics with three polynucleotide forms: plasmid DNA, oligodeoxynucleotide decoys, and small interfering RNA, in two cell types, H9C2(2-1) and Hela cells. The research program highlighted herein is driven by three specific goals: 1) to unravel the molecular-level interactions between structurally diverse yet analogous polymeric delivery vehicles and differing nucleic acid types and to correlate these interactions with the biological stability and mechanisms of the subsequent polyplexes, 2) to understand the interactions of these various polyplex types with cell surface glycosaminoglycans and compare polyplex structure to receptor selectivity and mechanisms of cellular uptake in two cell types. 3) To decipher the intracellular trafficking pathways in a spatial and temporal manner from uptake to the final destination for each polyplex form with the two model cell types. Public Health Relevance: The intracellular delivery of nucleic acids to study genetic and epigenetic processes associated with human health and disease offers unprecedented promise for revolutionizing biomedical research and drug development. However, it has been found that the nucleic acid delivery vehicle plays a central yet elusive role in dictating the efficacy, safety, mechanisms, and kinetics of gene regulation in a spatial and temporal manner. The goal of this project is to examine a wide range of novel nonviral glycopolymer-based delivery vehicles for their mechanistic pathways and kinetics of nucleic acid encapsulation and intracellular transport from the cell surface to their final intracellular destination with two models (cardiomyocytes and cancer cells) that represent the leading causes of disease and death.

Agency
National Institute of Health (NIH)
Institute
Office of The Director, National Institutes of Health (OD)
Type
NIH Director’s New Innovator Awards (DP2)
Project #
7DP2OD006669-02
Application #
8411640
Study Section
Special Emphasis Panel (ZGM1-NDIA-O (02))
Program Officer
Basavappa, Ravi
Project Start
2009-09-30
Project End
2014-06-30
Budget Start
2011-09-01
Budget End
2014-06-30
Support Year
2
Fiscal Year
2009
Total Cost
$1,846,709
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
555917996
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Boyle, William S; Senger, Kyle; Tolar, Jakub et al. (2017) Heparin Enhances Transfection in Concert with a Trehalose-Based Polycation with Challenging Cell Types. Biomacromolecules 18:56-67
Wu, Yaoying; Smith, Adam E; Reineke, Theresa M (2017) Lipophilic Polycation Vehicles Display High Plasmid DNA Delivery to Multiple Cell Types. Bioconjug Chem 28:2035-2040
Tolstyka, Zachary P; Phillips, Haley; Cortez, Mallory et al. (2016) Trehalose-Based Block Copolycations Promote Polyplex Stabilization for Lyophilization and in Vivo pDNA Delivery. ACS Biomater Sci Eng 2:43-55
Nasiri, Mohammadreza; Bertrand, Arthur; Reineke, Theresa M et al. (2014) Polymeric nanocylinders by combining block copolymer self-assembly and nanoskiving. ACS Appl Mater Interfaces 6:16283-8
Sprouse, Dustin; Reineke, Theresa M (2014) Investigating the effects of block versus statistical glycopolycations containing primary and tertiary amines for plasmid DNA delivery. Biomacromolecules 15:2616-28
Wu, Yaoying; Wang, Miao; Sprouse, Dustin et al. (2014) Glucose-containing diblock polycations exhibit molecular weight, charge, and cell-type dependence for pDNA delivery. Biomacromolecules 15:1716-26
Burke, Paul A; Pun, Suzie H; Reineke, Theresa M (2013) Advancing polymeric delivery systems amidst a nucleic acid therapy renaissance. ACS Macro Lett 2:928-934
Sizovs, Antons; Xue, Lian; Tolstyka, Zachary P et al. (2013) Poly(trehalose): sugar-coated nanocomplexes promote stabilization and effective polyplex-mediated siRNA delivery. J Am Chem Soc 135:15417-24
Xue, Lian; Ingle, Nilesh P; Reineke, Theresa M (2013) Highlighting the role of polymer length, carbohydrate size, and nucleic acid type in potency of glycopolycation agents for pDNA and siRNA delivery. Biomacromolecules 14:3903-15
Li, Haibo; Cortez, Mallory A; Phillips, Haley R et al. (2013) Poly(2 deoxy 2 methacrylamido glucopyranose) b Poly(methacrylate amine)s: Optimization of Diblock Glycopol ycations for Nucleic Acid Delivery. ACS Macro Lett 2:

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