Polyethylenimine/DNA complexes shielded by transferrin target gene expression to tumors after systemic application

Kircheis, Ralf; Wightman, Lionel; Schreiber, Alan D.; Robitza, B; Rössler, Vanessa; Kursa, Malgorzata; Wagner, Ernst

doi:10.1038/sj.gt.3301351

Cited by 331 publications

(257 citation statements)

References 44 publications

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“…With increasing N/P ratios even marked fetal mortality was observed, most probably caused by embolic events either in the livers or lungs of the injected animals because of the high electropositivity of the complexes leading to aggregation with blood components. 15 According to our results, linear PEI22 is better suited for in utero application than the branched forms of the polycation (PEI800 or PEI25). Best in vivo results were obtained with ligand-free PEI22/DNA complexes generated in the presence of salt.…”

Section: Discussionmentioning

confidence: 61%

“…This was surprising, since incorporation of the ligand into the complexes had previously been shown to be beneficial for postnatal in vivo approaches. 15 Previous reports have described much higher reporter gene expression in adult mouse lungs with salt-free PEI22/DNA complexes as compared to salt-containing formulations. 18,19 This was confirmed in our study.…”

Section: Discussionmentioning

confidence: 97%

“…The only exception was found with PEI22/DNA complexes in adult animals, where similar or even higher gene expression was found in the lungs compared to the liver, which is in agreement with previous studies showing particularly high gene transfer into lungs with PEI22/DNA complexes. 15,18,19 No reporter gene expression was detected in the kidneys or gonads of any of the mice analyzed (266 mice injected prenatally with different PEI/DNA complexes; 49 mice injected prenatally with different amounts of naked DNA and 78 postnatally injected mice (3-day-old and 8-week-old) with either PEI-complexed or naked DNA).…”

Section: Transgene Expression In Nontarget Organsmentioning

confidence: 99%

“…10,11 In the present study, the cationic polymer polyethylenimine (PEI) was used for intrauterine gene transfer. PEI's ability to condense and deliver DNA to cells has been extensively characterized in vivo, [12][13][14][15] but the applicability of this substance for prenatal gene transfer has not been addressed to date. A target organ that is particularly interesting for in utero gene delivery is the fetal liver.…”

Section: Introductionmentioning

confidence: 99%

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Nonviral gene transfer into fetal mouse livers (a comparison between the cationic polymer PEI and naked DNA)

et al. 2003

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We investigated the efficacy and safety of the cationic polymer polyethylenimine (PEI) as a potential tool for intrauterine gene delivery into livers of fetal mice in the last trimester of pregnancy (E17.5). Using luciferase as a reporter gene, transferrin-conjugated and ligand-free PEI/ DNA complexes (containing 3 mg DNA) with varying PEInitrogen/DNA-phosphate (N/P) ratios and different PEI forms, branched (800, 25 kDa) and linear (22 kDa), were compared with naked DNA. Transgene expression was measured 48 h after administration of PEI/DNA complexes or naked DNA. Highest luciferase activity (9.8 Â 10 3 relative light units (RLU)/mg of tissue protein) was observed with ligand-free PEI22/DNA mixtures at N/P 6.0. In addition, this formulation was associated with very low toxicity as compared to the other PEI/DNA-injected groups. Using b-galactosidase as a reporter gene, transfection of single, but also small, clusters of cells was demonstrated throughout the liver. Injection of 3 mg naked DNA resulted in an 11-fold lower transgene expression value (0.9 Â 10 3 RLU/mg of tissue protein) as compared to PEI22/DNA complexes. However, the administration of higher concentrated naked DNA (9 mg) into fetal livers yielded expression levels of 3.2 Â 10 4 RLU/mg of tissue protein, a more than three-fold increase compared to PEI22/DNA complexes. Furthermore, the gene transfer efficacy of concentrated naked DNA was approximately 40 times higher in fetuses than in adults (0.8 Â 10 3 RLU/mg of tissue protein), indicating that fetal tissue is especially amenable to the uptake and expression of naked DNA.

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Section: Discussionmentioning

confidence: 61%

Section: Discussionmentioning

confidence: 97%

Section: Transgene Expression In Nontarget Organsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Nonviral gene transfer into fetal mouse livers (a comparison between the cationic polymer PEI and naked DNA)

et al. 2003

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“…An exception to this is delivery to the lung, where it has been shown that large aggregating PEI/DNA particles passively target the lung following tail-vein injection. 23,24 For receptor-specific endocytosis, ligands such as epidermal growth factor (EGF) 25 and RGD peptide 26 have been utilized to increase cell specificity and overall uptake in vitro and in vivo. Though targeting often increases transfection, some experiments show that uptake that is both specific and efficient only occurs at a narrow window of charge ratios or not at all.…”

Section: Designing Polymeric Nanoparticles For Increased Uptake and Lmentioning

confidence: 99%

A Combinatorial Polymer Library Approach Yields Insight into Nonviral Gene Delivery

Green¹,

2008

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CONSPECTUSThe potential of gene therapy to benefit human health is tremendous because almost all human diseases have a genetic component, from untreatable monogenic disorders to cancer and heart disease. Unfortunately, a method for gene therapy that is both effective and safe has remained elusive. It has been said that "there are only three problems in gene therapy -delivery, delivery, and delivery." (quote from I. M. Verma in Jaroff, L. TIME, 1999; Jan 11).This Account describes an alternative strategy to viral gene delivery: the design of biodegradable polymers that are able to deliver DNA like a synthetic virus. Using high-throughput synthesis and screening techniques, we have created libraries of over 2000 structurally unique poly(β-amino esters) (PBAEs). PBAEs are formed by the conjugate addition of amines to diacrylates. These biomaterials are promising for nonviral gene delivery due to their ability to condense plasmid DNA into small and stable nanoparticles and their ability to promote cellular uptake and endosomal escape. Our laboratory has iteratively improved PBAE nanoparticles through polymer end modifications and nanoparticle coatings. Lead PBAEs have high gene delivery efficacy and low cytotoxicity both in vitro and in vivo.Certain polymer structural characteristics are important for effective gene delivery. The best PBAEs are linear polymers of ~10 kDa that contain hydroxyl side chains and primary amine end groups. These polymers bind DNA to form nanoparticles that are small (<200 nm) and stable and have near-neutral ζpotential in the presence of serum-containing media. Lead PBAEs also contain tertiary amines that can buffer the low pH environment of endosomes and facilitate escape of polymer/DNA particles into the cytoplasm.Diamine end-modified 1,4-butanediol diacrylate-co-5-amino-1-pentanol polymers (C32) bind DNA more tightly and form smaller nanoparticles than other PBAEs. These nanoparticles also have higher cellular uptake and the best gene expression of all gene delivery polymers in the library. These polymers are more effective for gene delivery than top commercially available nonviral vectors including jet-PEI and Lipofectamine 2000 and are comparable to adenovirus for in vitro gene delivery to human primary cells. In vivo, these PBAE/ DNA particles are promising as cancer therapeutics. This Account summarizes the results of our laboratory in using a combinatorial polymer library approach to elucidate polymer structure/function relationships and enable the development of polymeric gene delivery nanoparticles with viral-like efficacy. Nonviral gene delivery systems make use of physical methods or a synthetic chemical vector or both to deliver the gene of interest. Limitations with viral approaches, including small cargo capacity, resistance to repeated infection, difficulty in production and quality control, and low safety, can be potentially overcome with a nonviral approach. Originally, nonviral gene delivery was simply delivery of naked plasmid DNA. To improve transfection, physical str...

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Gene‐Delivery Polymers

Pack

2002

Encyclopedia of Polymer Science and Technology

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Human gene therapy promises to provide unparalleled treatments for afflictions ranging from cystic fibrosis to cardiovascular disease to cancer. The development of safe and efficient methods for delivery of genetic material to specific cells within the body has been a limiting factor in implementation of this technology. Delivery methodologies have included the use of recombinant viruses and synthetic materials, especially cationic polymers. While viruses are by far the most efficient delivery vehicles currently available, polymers exhibit several properties—including very flexible chemistry, potential for biocompatibility, simplicity, and inexpensive synthesis—that make them excellent candidates for the gene delivery vehicles of the future. However, polymers are typically several orders of magnitude less efficient than viruses and are generally considered unacceptable for human gene therapy. Much work is underway to design and synthesize improved polymers for gene delivery applications. This article summarizes the important criteria for an ideal gene delivery vehicle in terms of the potential barriers that must be overcome. Further, the current state of the polymer‐mediated gene delivery field is reviewed and several new classes of polymers, specifically designed for human gene delivery, are described.

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Polyethylenimine/DNA complexes shielded by transferrin target gene expression to tumors after systemic application

Cited by 331 publications

References 44 publications

Nonviral gene transfer into fetal mouse livers (a comparison between the cationic polymer PEI and naked DNA)

Nonviral gene transfer into fetal mouse livers (a comparison between the cationic polymer PEI and naked DNA)

A Combinatorial Polymer Library Approach Yields Insight into Nonviral Gene Delivery

Gene‐Delivery Polymers

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