CL22 – a novel cationic peptide for efficient transfection of mammalian cells

Haines, A. M. R.; Irvine, Alistair S.; Mountain, Andrew; Charlesworth, Jon; Farrow, Neil A.; Husain, Rhonda; Hyde, Helena; Ketteringham, Helen; McDermott, R. H.; Mulcahy, A. F.; Mustoe, Tracey; Reid, Sophie C.H.; Rouquette, Magali; Shaw, J.C.; Thatcher, David R.; Welsh, John H.; Williams, DE; Zauner, Wolfgang; Phillips, Rob

doi:10.1038/sj.gt.3301314

Cited by 40 publications

(21 citation statements)

References 29 publications

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“…Complexing with aBNBD3 may have condensed the DNA, making its size and shape more attractive for cellular uptake (55,56). In addition, aBNBD3 may have increased the binding and entry of the DNA into cells across the negatively charged cell membranes, as has been reported for cationic liposomes (57) and cationic peptides (54,58).…”

Section: Discussionmentioning

confidence: 95%

Immunogenicity of a Bovine Herpesvirus 1 Glycoprotein D DNA Vaccine Complexed with Bovine Neutrophil Beta-Defensin 3

Mackenzie-Dyck

Latimer

Atanley

et al. 2014

Clin. Vaccine Immunol.

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Protective efficacy against bovine herpesvirus 1 (BoHV-1) has been demonstrated to be induced by a plasmid encoding bovine neutrophil beta-defensin 3 (BNBD3) as a fusion construct with truncated glycoprotein D (tgD). However, in spite of the increased cell-mediated immune responses induced by this DNA vaccine, the clinical responses of BoHV-1-challenged cattle were not reduced over those observed in animals vaccinated with the plasmid encoding tgD alone; this might have been because the vaccine failed to improve humoral responses. We hypothesized that an alternative vaccine design strategy that utilized the DNA vaccine pMASIA-tgD as a complex with BNBD3 might improve humoral responses while maintaining robust Th1-type cell-mediated responses. C57BL/6 mice were vaccinated with pMASIA-tgD complexed with 0, 0.01875, 0.1875, or 1.875 nmol of a stable synthesized analog of BNBD3 (aBNBD3). The best results were seen in mice immunized with the vaccine composed of pMASIA-tgD complexed to 0.1875 nmol aBNBD3. In this group, humoral responses were improved, as evidenced by increased virus neutralization, tgD-specific early IgG1, and later IgG2a titers, while the strong cell-mediated immune responses, measured based on specific gamma interferon (IFN-␥)-secreting cells, were maintained relative to pMASIA-tgD. Modulation of the immune response might have been due in part to the effect of BNBD3 on dendritic cells (DCs). In vitro studies showed that murine bone marrow-derived DCs (BMDCs) pretreated with aBNBD3 were activated, as evidenced by CD11c downregulation, and were functionally mature, as shown by increased allostimulatory ability. Native, synthetic, and analog forms of BNBD3 were equally capable of inducing functional maturation of BMDCs.

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Section: Discussionmentioning

confidence: 95%

Immunogenicity of a Bovine Herpesvirus 1 Glycoprotein D DNA Vaccine Complexed with Bovine Neutrophil Beta-Defensin 3

Mackenzie-Dyck

Latimer

Atanley

et al. 2014

Clin. Vaccine Immunol.

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“…5 Many companies have developed lipofection systems with improved transduction rates. 1,2 Some of these lipofection systems, especially Fugene6t, however, are not cGMP-conform. 3 For these reasons, we tested our cGMP-quality lipids, CCQ22 and CCQ32, 3 in breast and ovarian cancer substrates.…”

Section: Discussionmentioning

confidence: 99%

“…In contrast, nonviral vectors show lower immunogenicity and easier cGMPproduction. 1 Therefore, an attractive concept is the development of highly efficient nonviral, liposomal vector systems with acceptable toxicity profiles.…”

mentioning

confidence: 99%

Novel cGMP liposomal vectors mediate efficient gene transfer

et al. 2003

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Viral vector systems are the most commonly used gene transfer tools for clinical gene therapy. However, lipofection systems are potential alternatives because of lower immunogenicity and easier cGMP production, but in vivo stability and transduction efficacy need to be improved. Therefore, we investigated gene transduction efficiency of our novel cGMP cationic lipids, CCQ22 and CCQ32, by FACS analysis. Toxicity analysis was performed to determine the cytotoxic side effects of the novel lipids. To evaluate the stability of the compounds in the context of local delivery to patients with intraperitoneally metastatic ovarian cancer, gene transfer was also tested in the presence of malignant ascites. Our novel cGMP standard lipids mediated gene transfer rates of more than 50%. However, for most cell lines cytotoxic side effects were similar to our reference lipofection system. In general, ascites had no major influence on gene transduction rates with the novel lipids. Our results suggest that CCQs may compare favorably with commercially available lipofection systems. These promising results facilitate further analysis of the compounds.

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“…The pDNA 12 mobility was visualized by ethidium bromide staining. 13 To evaluate the ability of the vector in protecting pDNA from endonucleases, complexes 14 were formed at an optimal N:P 10 in a microfuge tube and incubated with serum for 90 minutes. 15 Subsequently, sodium dodecyl sulfate was added (10%) to the tubes followed by heating to 95°C 16 for 5 minutes to release the pDNA from the vector.…”

Section: Recognition Of Cathepsin D Substrate By Cathepsin D Protease 23mentioning

confidence: 99%

“…Because of the cyclic characteristic of the targeting peptide, the DBV was engineered to 16 stably exist as a monomer and dimer (Fig 1a). A cathepsin D substrate is also engineered in the 17 vector structure to facilitate dissociation of the targeting peptide from the vector inside 18 endosomes where cathepsin D is abundant [13]. The proposed model for the intracellular 19 trafficking of the vector is shown in Fig.…”

Section: Introductionmentioning

confidence: 99%

Development of a Nature-Inspired Vector for Targeted Systemic Breast Cancer Gene Therapy

Hatefi¹

2008

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. Send comments regarding this burden estimate or any other aspect of this collection of information, including suggestions for reducing this burden to Department of Defense, Washington Headquarters Services, Directorate for Information Operations and Reports (0704-0188), 1215 Jefferson Davis Highway, Suite 1204, Arlington, VA 22202-4302. Respondents should be aware that notwithstanding any other provision of law, no person shall be subject to any penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. (From -To) REPORT DATE (DD-MM-YYYY) 01/09/2008 REPORT TYPE Annual Report DATES COVERED SPONSORING / MONITORING AGENCY NAME(S) AND ADDRESS(ES) 10. SPONSOR/MONITOR'S ACRONYM(S) U.S. Army Medical Research and Materiel Command Fort Detrick, Maryland 21702-5012 SPONSOR/MONITOR'S REPORT NUMBER(S) DISTRIBUTION / AVAILABILITY STATEMENTApproved for public release; distribution unlimited SUPPLEMENTARY NOTES ABSTRACT:The purpose of this research was to develop a gene delivery system that can target breast cancer cells specifically and transfect them efficiently. Using component integration approach, several subsystems of diverse biological origin were integrated onto a modular platform in order to carry diverse functions of an efficient gene delivery vehicle. A biomimetic vector was genetically engineered to contain at precise locations: a) an adenovirus µ peptide to condense pDNA into nanosize particles, b) a combinatorially-screened synthetic peptide to target breast cancer cells and enhance internalization of nanoparticles, c) a pH-responsive synthetic fusogenic peptide to disrupt endosome membranes and facilitate escape of the nanoparticles into the cytosol, and d) a nuclear localization signal from human immuno-deficiency virus for microtubule mediated transfer of genetic material to the nucleus. The vector was characterized physicochemically and biologically and the results demonstrated that a fully functional vector can be engineered to target breast cancer cells with high specificity, mimic virus characteristics and overcome the biological barriers associated with targeted gene transfer. Conclusions……………………………………………………………………………3References……………………………………………………………………………..4Appendices…………………………………………………………………………….41 IntroductionThe overall purpose of this project was to develop a new generation of programmable non-viral vectors to target and transfect breast cancer cells efficiently. The rationale is that a multidomain designer vector based on a One-Domain-One-Function architecture can effectively perform an array of functions including a) self assembly into vector/DNA nanoparticles i.e., packaging the therapeutic ge...

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CL22 – a novel cationic peptide for efficient transfection of mammalian cells

Cited by 40 publications

References 29 publications

Immunogenicity of a Bovine Herpesvirus 1 Glycoprotein D DNA Vaccine Complexed with Bovine Neutrophil Beta-Defensin 3

Immunogenicity of a Bovine Herpesvirus 1 Glycoprotein D DNA Vaccine Complexed with Bovine Neutrophil Beta-Defensin 3

Novel cGMP liposomal vectors mediate efficient gene transfer

Development of a Nature-Inspired Vector for Targeted Systemic Breast Cancer Gene Therapy

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