Publications by authors named "Yong Serk Park"

33 Publications

Identification of genes involved in neuronal cell death and recovery over time in rat axotomy and neurorrhaphy models through RNA sequencing.

Mol Cell Neurosci 2021 Jun 5;113:103628. Epub 2021 May 5.

Regeneration Medicine Research Center, Yonsei University Wonju College of Medicine, Wonju, Republic of Korea. Electronic address:

Facial nerves are frequently injured during cosmetic or other types of facial surgery. However, information on the genes involved in the damage and recovery of the facial nerves is limited. Here, we aimed to identify the genes affected by facial nerve injury and repair using next-generation sequencing. We established a rat axotomy model and a parallel epineurial neurorrhaphy model, in which gene expression was analyzed from 3 days to 8 weeks after surgery. We discovered that ARRB1, SGK1, and GSK3B genes associated with neuronal cell death were upregulated in the axotomy model. In contrast, MFRP, MDK, and ACE genes involved in neural recovery and regeneration exhibited higher expression in the neurorrhaphy model. In the present study, the analysis of the big data obtained from the next-generation sequencing (RNA-seq) technology reveals that the expression of genes involved in neuronal cell death is induced during nerve damage, and those associated with neural recovery are more abundantly expressed during repair processes. These results are considered to be useful for the establishment of the treatment of related diseases and basic research in various neuroscience fields by utilizing damage and recovery mechanism of facial nerves.
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http://dx.doi.org/10.1016/j.mcn.2021.103628DOI Listing
June 2021

Development of HER2-Specific Aptamer-Drug Conjugate for Breast Cancer Therapy.

Int J Mol Sci 2020 Dec 21;21(24). Epub 2020 Dec 21.

Biois Co., Ltd., Seoul 08390, Korea.

In this study, HER2 RNA aptamers were conjugated to mertansine (DM1) and the anti-cancer effectiveness of the conjugate was evaluated in HER2-overexpressing breast cancer models. The conjugate of HER2 aptamer and anticancer drug DM1 (aptamer-drug conjugate, ApDC) was prepared and analyzed using HPLC and mass spectrometry. The cell-binding affinity and cytotoxicity of the conjugate were determined using confocal microscopy and WST-1 assay. The in vivo anti-tumoral efficacy of ApDC was also evaluated in mice carrying BT-474 breast tumors overexpressing HER2. The synthesized HER2-specific RNA aptamers were able to specifically and efficiently bind to HER-positive BT-474 breast cancer cells, but not to HER2-negative MDA-MB-231 breast cancer cells. Also, the HER2-specific ApDC showed strong toxicity to the target cells, BT-474, but not to MDA-MB-231 cells. According to the in vivo analyses drawn from the mouse xenografts of BT-747 tumor, the ApDC was able to more effectively inhibit the tumor growth. Compared to the control group, the mice treated with the ApDC showed a significant reduction of tumor growth. Besides, any significant body weight losses or hepatic toxicities were monitored in the ApDC-treated mice. This research suggests the HER2 aptamer-DM1 conjugate as a target-specific anti-cancer modality and provides experimental evidence supporting its enhanced effectiveness for HER2-overexpressing target tumors. This type of aptamer-conjugated anticancer drug would be utilized as a platform structure for the development of versatile targeted high-performance anticancer drugs by adopting the easy deformability and high affinity of aptamers.
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http://dx.doi.org/10.3390/ijms21249764DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7767363PMC
December 2020

Development of a Theranostic Convergence Bioradiopharmaceutical for Immuno-PET Based Radioimmunotherapy of L1CAM in Cholangiocarcinoma Model.

Clin Cancer Res 2019 10 23;25(20):6148-6159. Epub 2019 Jul 23.

Division of RI Application, Korea Institute of Radiological and Medical Sciences, Seoul, South Korea.

Purpose: Cholangiocarcinoma is a malignancy of bile duct with a poor prognosis. Conventional chemotherapy and radiotherapy are generally ineffective, and surgical resection is the only curative treatment for cholangiocarcinoma. L1-cell adhesion molecule (L1CAM) has been known as a novel prognostic marker and therapeutic target for cholangiocarcinoma. This study aimed to evaluate the feasibility of immuno-PET imaging-based radioimmunotherapy using radiolabeled anti-L1CAM antibody in cholangiocarcinoma xenograft model.

Experimental Design: We prepared a theranostic convergence bioradiopharmaceutical using chimeric anti-L1CAM antibody (cA10-A3) conjugated with 1,4,7-triazacyclononane-1,4,7-triacetic acid (NOTA) chelator and labeled with Cu or Lu and evaluated the immuno-PET or SPECT/CT imaging and biodistribution with Cu-/Lu-cA10-A3 in various cholangiocarcinoma xenograft models. Therapeutic efficacy and response monitoring were performed by Lu-cA10-A3 and F-FDG-PET, respectively, and immunohistochemistry was done by TUNEL and Ki-67.

Results: Radiolabeled cA10-A3 antibodies specifically recognized L1CAM , clearly visualized cholangiocarcinoma tumors in immuno-PET and SPECT/CT imaging, and differentiated the L1CAM expression level in cholangiocarcinoma xenograft models. Lu-cA10-A3 (12.95 MBq/100 μg) showed statistically significant reduction in tumor volumes ( < 0.05) and decreased glucose metabolism ( < 0.01). IHC analysis revealed Lu-cA10-A3 treatment increased TUNEL-positive and decreased Ki-67-positive cells, compared with saline, cA10-A3, or Lu-isotype.

Conclusions: Anti-L1CAM immuno-PET imaging using Cu-cA10-A3 could be translated into the clinic for characterizing the pharmacokinetics and selecting appropriate patients for radioimmunotherapy. Radioimmunotherapy using Lu-cA10-A3 may provide survival benefit in L1CAM-expressing cholangiocarcinoma tumor. Theranostic convergence bioradiopharmaceutical strategy would be applied as imaging biomarker-based personalized medicine in L1CAM-expressing patients with cholangiocarcinoma.
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http://dx.doi.org/10.1158/1078-0432.CCR-19-1157DOI Listing
October 2019

Anti-EGF Receptor Aptamer-Guided Co-Delivery of Anti-Cancer siRNAs and Quantum Dots for Theranostics of Triple-Negative Breast Cancer.

Theranostics 2019 25;9(3):837-852. Epub 2019 Jan 25.

Department of Biomedical Laboratory Science, Yonsei University, Wonju, Republic of Korea.

Many aptamers have been evaluated for their ability as drug delivery vehicles to target ligands, and a variety of small interfering RNAs (siRNAs) have been tested for their anti-cancer properties. However, since these two types of molecules have similar physicochemical properties, it has so far been difficult to formulate siRNA-encapsulating carriers guided by aptamers. Here, we propose aptamer-coupled lipid nanocarriers encapsulating quantum dots (QDs) and siRNAs for theragnosis of triple-negative breast cancer (TNBC). Hydrophobic QDs were effectively incorporated into lipid bilayers, and then therapeutic siRNAs were complexed with QD-lipid nanocarriers (QLs). Finally, anti-EGFR aptamer-lipid conjugates were inserted into the QLs for TNBC targeting (aptamo-QLs). TNBC-targeting aptamo-QLs were directly compared to anti-EGFR antibody-coupled immuno-QLs. The delivery of therapeutic siRNAs and QDs to target cells was assessed by flow cytometry and confocal microscopy. The targeting of siRNAs to tumors and their therapeutic efficacy were evaluated in mice carrying MDA-MB-231 tumors. Both types of EGFR-targeting QLs showed enhanced delivery to target cancer cells, resulting in more effective gene silencing and enhanced tumor imaging compared to non-targeting control QLs. Moreover, combinatorial therapy with Bcl-2 and PKC-ι siRNAs loaded into the anti-EGFR QLs was remarkably effective in inhibiting tumor growth and metastasis. In general, the aptamo-QLs showed competitive delivery and therapeutic efficacy compared to immuno-QLs under the same experimental conditions. Our results show that the anti-EGFR aptamer-guided lipid carriers may be a potential theranostic delivery vehicle for RNA interference and fluorescence imaging of TNBCs.
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http://dx.doi.org/10.7150/thno.30228DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6376474PMC
December 2019

PET Imaging Biomarkers of Anti-EGFR Immunotherapy in Esophageal Squamous Cell Carcinoma Models.

Cells 2018 Oct 27;7(11). Epub 2018 Oct 27.

Division of RI Application, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul 01812, Korea.

Epidermal growth factor receptor (EGFR) is overexpressed and considered as a proper molecular target for diagnosis and targeted therapy of esophageal squamous cell carcinoma (ESCC). This study evaluated the usefulness of PET imaging biomarkers with Cu-PCTA-cetuximab and F-FDG-PET for anti-EGFR immunotherapy in ESCC models. EGFR status and glucose metabolism by cetuximab treatment were evaluated using Cu-PCTA-cetuximab and F-FDG-PET, respectively. Therapeutic responses with imaging biomarkers were confirmed by western blot and immunohistochemistry. TE-4 and TE-8 tumors were clearly visualized by Cu-PCTA-cetuximab, and EGFR expression on TE-8 tumors showed 2.6-fold higher uptake than TE-4. Tumor volumes were markedly reduced by cetuximab in TE-8 tumor (92.5 ± 5.9%), but TE-4 tumors were refractory to cetuximab treatment. The SUVs in Cu-PCTA-cetuximab and F-FDG-PET images were statistically significantly reduced by cetuximab treatment in TE-8 but not in TE-4. Cu-PCTA-cetuximab and F-FDG-PET images were well correlated with EGFR and pAkt levels. Cu-PCTA-cetuximab immuno-PET had a potential for determining EGFR level and monitoring therapeutic response by anti-EGFR therapy. F-FDG-PET was also attractive for monitoring efficacy of anti-EGFR therapy. In conclusion, PET imaging biomarkers may be useful for selecting patients that express target molecules and for monitoring therapeutic efficacy of EGFR-targeted therapy in ESCC patients.
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http://dx.doi.org/10.3390/cells7110187DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6262544PMC
October 2018

Anti-EGFR lipid micellar nanoparticles co-encapsulating quantum dots and paclitaxel for tumor-targeted theranosis.

Nanoscale 2018 Nov 11;10(41):19338-19350. Epub 2018 Oct 11.

Department of Biomedical Laboratory Science, Yonsei University, Wonju, Republic of Korea.

Cancer theranosis is an emerging field of personalized medicine which enables individual anti-cancer treatment by monitoring the therapeutic responses of cancer patients. Based on a consideration of the nano-bio interactions related to the blood circulation of systemically administered nanoparticles in humans, as well as extravasation and active targeting, lipid micellar nanoparticles were co-loaded with paclitaxel (PTX) and quantum dots (QDs) to generate a theranostic delivery vehicle. To provide with a tumor-targeting capability, either an antibody or an aptamer against the epidermal growth factor receptor (EGFR) was conjugated to the micelle surface. The QD-containing micelles (QDMs), antibody-coupled QDMs (immuno-QDMs), and aptamer-coupled QDMs (aptamo-QDMs) were able to effectively circulate in blood for at least 8 h when administered intravenously into mice bearing EGFR-positive LS174T tumor xenografts. In vivo fluorescence imaging and a bio-distribution study showed that both the immuno-QDMs and aptamo-QDMs were largely localized in the tumor tissue. The tumor targeting capability enhanced the therapeutic efficacy of PTX for the target cancer cells. Both the immuno-PTX-QDMs and the aptamo-PTX-QDMs caused a stronger inhibition of LS174T tumor growth in mice, compared to the non-targeted PTX-QDMs. These results suggest that the anti-EGFR immuno-PTX-QDMs and anti-EGFR aptamo-PTX-QDMs could be utilized as a tumor-targeted theranostic delivery system for cancer treatment in the clinic.
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http://dx.doi.org/10.1039/c8nr05099fDOI Listing
November 2018

Tumor-specific delivery of therapeutic siRNAs by anti-EGFR immunonanoparticles.

Int J Nanomedicine 2018 27;13:4817-4830. Epub 2018 Aug 27.

Department of Biomedical Laboratory Science, Yonsei University, Wonju, Republic of Korea,

Background: Efficient target-specific siRNA delivery has always been a primary concern in the field of siRNA clinical application.

Purpose: In this study, four different types of anti-epidermal growth factor receptor (EGFR) antibody-conjugated immunonanoparticles were prepared and tested for cancer cell-targeted therapeutic siRNA delivery.

Materials And Methods: The prepared nanoparticles encapsulating siRNAs were character-ized by gel retardation and particle analysis using a Zetasizer. In vitro transfection and reduction of target genes, vimentin and JAK3, were determined using quantitative reverse transcription polymerase chain reaction. In vivo tumor targeting and antitumoral efficacies of the nanoparticles were evaluated in mice carrying tumors.

Results: Among these immunonanoparticles, anti-EGFR immunolipoplexes and immunoviroplexes exhibited remarkable cell binding and siRNA delivery to EGFR-expressing tumor cells compared to immunoliposomes and immunovirosomes. Especially, the anti-EGFR immunoviroplexes exhibited the most efficient siRNA transfection to target tumor cells. Therefore, antitumoral vimentin and Janus kinase-3 siRNAs were loaded in the anti-EGFR immunolipoplexes and immunoviroplexes, which were tested in mice carrying SK-OV-3 tumor xenografts. In fact, the therapeutic siRNAs were efficiently delivered to the tumor tissues by both delivery vehicles, resulting in significant inhibition of tumor growth. Moreover, administration of doxorubicin in combination with anti-EGFR immunoviroplexes resulted in remarkable and synergistic tumor growth inhibition.

Conclusion: This study provides experimental proof that cancer cell-targeted immunoviroplexes are an efficient siRNA delivery system for cancer therapy. Moreover, this study also suggests that a combination of conventional chemotherapy and tumor-directed anticancer siRNA therapy would be a better modality for cancer treatment.
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http://dx.doi.org/10.2147/IJN.S161932DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6118344PMC
October 2018

Cancer-targeted Nucleic Acid Delivery and Quantum Dot Imaging Using EGF Receptor Aptamer-conjugated Lipid Nanoparticles.

Sci Rep 2017 08 25;7(1):9474. Epub 2017 Aug 25.

Department of Biomedical Laboratory Science, Yonsei University, Wonju, Republic of Korea.

Co-application of fluorescent quantum dot nanocrystals and therapeutics has recently become a promising theranostic methodology for cancer treatment. We developed a tumor-targeted lipid nanocarrier that demonstrates notable efficacy in gene delivery as well as tumor bio-imaging. Coupling of aptamer molecules against the EGF receptor (EGFR) to the distal termini of lipid nanoparticles provided the carrier with tumor-specific recognition capability. The cationic lipid component, referred to as O,O'-dimyristyl-N-lysyl glutamate (DMKE), was able to effectively complex with anionic small-interfering RNA (siRNA). The hydrophobic quantum dots (Q-dots) were effectively incorporated in hydrophobic lipid bilayers at an appropriate Q-dot to lipid ratio. In this study, we optimized the liposomal formula of aptamer-conjugated liposomes containing Q-dots and siRNA molecules (Apt-QLs). The anti-EGFR Apt-QLs exhibited remarkable EGFR-dependent siRNA delivery as well as fluorescence imaging, which were analyzed in cultured cancer cells and tumor xenografts in mice. These results imply that the formulation of Apt-QLs could be widely utilized as a carrier for tumor-directed gene delivery and bio-imaging.
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http://dx.doi.org/10.1038/s41598-017-09555-wDOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5573382PMC
August 2017

Anti-EGFR immunonanoparticles containing IL12 and salmosin genes for targeted cancer gene therapy.

Int J Oncol 2016 Sep 7;49(3):1130-8. Epub 2016 Jul 7.

Department of Biomedical Laboratory Science, Yonsei University, Wonju, Republic of Korea.

Tumor-directed gene delivery is of major interest in the field of cancer gene therapy. Varied functionalizations of non-viral vectors have been suggested to enhance tumor targetability. In the present study, we prepared two different types of anti-EGF receptor (EGFR) immunonanoparticles containing pDNA, neutrally charged liposomes and cationic lipoplexes, for tumor-directed transfection of cancer therapeutic genes. Even though both anti-EGFR immunonanoparticles had a high binding affinity to the EGFR-positive cancer cells, the anti-EGFR immunolipoplex formulation exhibited approximately 100-fold higher transfection to the target cells than anti-EGFR immunoliposomes. The lipoplex formulation also showed a higher transfection to SK-OV-3 tumor xenografts in mice. Thus, IL12 and/or salmosin genes were loaded in the anti-EGFR immunolipoplexes and intravenously administered to mice carrying SK-OV-3 tumors. Co-transfection of IL12 and salmosin genes using anti-EGFR immunolipoplexes significantly reduced tumor growth and pulmonary metastasis. Furthermore, combinatorial treatment with doxorubicin synergistically inhibited tumor growth. These results suggest that anti-EGFR immunolipoplexes containing pDNA encoding therapeutic genes could be utilized as a gene-transfer modality for cancer gene therapy.
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http://dx.doi.org/10.3892/ijo.2016.3619DOI Listing
September 2016

Sendai viroplexes for epidermal growth factor receptor-directed delivery of interleukin-12 and salmosin genes to cancer cells.

J Gene Med 2016 Jul;18(7):112-23

Department of Biomedical Laboratory Science, Yonsei University, Wonju, Republic of Korea.

Background: The effective delivery of therapeutic genes to target cells has been a fundamental goal in cancer gene therapy because of its advantages with respect to both safety and transfection efficiency. In the present, study we describe a tumor-directed gene delivery system that demonstrates remarkable efficacy in gene delivery and minimizes the off-target effects of gene transfection.

Methods: The system consists of a well-verified cationic O,O'-dimyristyl-N-lysyl glutamate (DMKE), Sendai virus fusion (F) protein and hemagglutinin-neuraminidase (HN) protein, referred to as cationic Sendai F/HN virosomes. To achieve tumor-specific recognition, anti-epidermal growth factor (EGF) receptor antibody was coupled to the surface of the virosomes containing interleukin-12 (IL-12) and/or salmosin genes that have potent anti-angiogenetic functions.

Results: Among the virosomal formulations, the anti-EGF receptor (EGFR) viroplexes, prepared via complexation of plasmid DNA (pDNA) with cationic DMKE lipid, exhibited more efficient gene transfection to tumor cells over-expressing EGF receptors compared to the neutrally-charged anti-EGFR virosomes encapsulating pDNA. In addition, the anti-EGFR viroplexes with IL-12 and salmosin genes exhibited the most effective therapeutic efficacy in a mouse tumor model. Especially when combined with doxorubicin, transfection of the two genes via the anti-EGFR viroplexes exhibited an enhanced inhibitory effect on tumor growth and metastasis in lungs.

Conclusions: The results of the present study suggest that anti-EGFR viroplexes can be utilized as an effective strategy for tumor-directed gene delivery. Copyright © 2016 John Wiley & Sons, Ltd.
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http://dx.doi.org/10.1002/jgm.2884DOI Listing
July 2016

Immuno-PET Imaging and Radioimmunotherapy of 64Cu-/177Lu-Labeled Anti-EGFR Antibody in Esophageal Squamous Cell Carcinoma Model.

J Nucl Med 2016 07 25;57(7):1105-11. Epub 2016 Feb 25.

Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences (KIRAMS), Seoul, South Korea Department of Nuclear Medicine, KIRAMS, Seoul, South Korea.

Unlabelled: Immuno-PET provides valuable information about tumor location, phenotype, susceptibility to therapy, and treatment response, especially to targeted radioimmunotherapy. In this study, we prepared antiepidermal growth factor receptor (EGFR) antibody via identical chelator, 3,6,9,15-tetraazabicyclo[9.3.1]-pentadeca-1(15),11,13-trience-3,6,9,-triacetic acid (PCTA), labeled with (64)Cu or (177)Lu to evaluate the EGFR expression levels using immuno-PET and the feasibility of radioimmunotherapy in an esophageal squamous cell carcinoma (ESCC) model.

Methods: Cetuximab was conjugated with p-SCN-Bn-PCTA and radiolabeled with (64)Cu or (177)Lu. In vitro EGFR expression levels were determined and compared using flow cytometry and cell binding assay. In vivo EGFR expression levels were evaluated via immuno-PET imaging of (64)Cu-cetuximab and biodistribution analysis. Micro-SPECT/CT imaging, biodistribution, and radioimmunotherapy studies of (177)Lu-cetuximab were performed in the ESCC model. Therapeutic responses were monitored using (18)F-FDG PET and immunohistochemical staining.

Results: (64)Cu- or (177)Lu-labeled antibodies showed high radiolabeling yield (>98%), stability (>90%), and favorable immunoreactivity. In vitro EGFR status measured by cell binding assay was correlated with the flow cytometry data. Immuno-PET, micro-SPECT/CT, and biodistribution demonstrated specific uptake in ESCC tumors depending on the EGFR expression levels. Tumor accumulation of (64)Cu- and (177)Lu-cetuximab was peaked at 48 and 120 h, respectively. Radioimmunotherapy with (177)Lu-cetuximab showed significant inhibition of tumor growth (P < 0.01) and marked reduction of (18)F-FDG SUV compared with that of control (P < 0.05). Terminal deoxynucleotidyl transferase dUTP nick-end labeling positivity and Ki-67 staining indices increased and decreased, respectively, in the radioimmunotherapy group compared with other groups (P < 0.01).

Conclusion: (64)Cu-cetuximab immuno-PET represented EGFR expression levels in ESCC tumors, and (177)Lu-cetuximab radioimmunotherapy effectively inhibited the tumor growth. The diagnostic and therapeutic convergence radiopharmaceutical (64)Cu-/(177)Lu-PCTA-cetuximab may be useful as a diagnostic tool in patient selection and a potent radioimmunotherapy agent in EGFR-positive ESCC tumors.
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http://dx.doi.org/10.2967/jnumed.115.167155DOI Listing
July 2016

RNA aptamer-conjugated liposome as an efficient anticancer drug delivery vehicle targeting cancer cells in vivo.

J Control Release 2014 Dec 24;196:234-42. Epub 2014 Oct 24.

Department of Bioscience and Biotechnology, Konkuk University, Seoul 143-701, Republic of Korea. Electronic address:

To minimize the systemic toxicity prevalent to chemotherapeutics, we designed a novel anticancer drug-encapsulating liposome conjugated with an RNA aptamer specific to the prostate specific membrane antigen (PSMA), which is expressed on the surface of prostate cancer cells. The RNA aptamer-conjugated liposome, termed an aptamosome, was prepared by the post-insertion method, in which RNA aptamer-conjugated micelles were inserted into a liposome. These nanosized (90-100 nm) aptamer-conjugated liposomes specifically bind to LNCaP prostate epithelial cells that express PSMA and thus cause the nanoparticles to have significantly enhanced in vitro cellular binding and uptake as compared with nontargeted nanoparticles that lack the PSMA aptamer. Aptamosomes encapsulated with the anticancer drug doxorubicin (Dox) were significantly more toxic to the targeted LNCaP cells than to nontargeted cancer cells. Dox-encapsulating aptamosomes administered to LNCaP xenograft nude mice were selectively retained in tumor tissue. We also demonstrated in vivo anticancer efficacy of the Dox-encapsulating PSMA-aptamosomes on tumor size regression in LNCaP xenograft mice. We suggest that the encapsulation of toxic chemicals with aptamer-conjugated liposomes will enable the use of these bioconjugates in clinical practice with fewer side effects.
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http://dx.doi.org/10.1016/j.jconrel.2014.10.018DOI Listing
December 2014

Synergistic antitumoral effect of IL-12 gene cotransfected with antiangiogenic genes for angiostatin, endostatin, and saxatilin.

Oncol Res 2013 ;21(4):209-16

Department of Biomedical Laboratory Science, Korea Nazarene University, Cheonan, Korea.

Previously, it was reported that the cotransfection of angiostatin K1-3, endostatin, and saxatilin genes using cationic liposomes significantly inhibited tumor progression. IL-12 is a well-known immune modulator that promotes Th1-type antitumor immune responses and also induces antiangiogenic effects. In this study, we have examined the antitumoral function of the IL-12 gene cotransfected with antiangiogenic genes for angiostatin K1-3, endostatin, and saxatilin by O,O'-dimyristyl-N-lysyl glutamate (DMKE) cationic liposomes in a mouse tumor model. According to our results, the administration of the IL-12 gene or the genes for angiostatin K1-3, endostatin, and saxatilin exhibited effective inhibition of B16BL6 melanoma growth in mice. In particular, intravenous administration of the IL-12 gene along with intratumoral administration of the three antiangiogenic genes synergistically inhibited the B16BL6 tumor growth. These results suggest that systemically expressed IL-12 enhances antitumoral efficacy of locally expressed antiangiogenic proteins.
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http://dx.doi.org/10.3727/096504014X13907540404798DOI Listing
December 2014

Sendai F/HN viroplexes for efficient transfection of leukemic T cells.

Yonsei Med J 2013 Sep;54(5):1149-57

Department of Biomedical Laboratory Science, Yonsei University, 1 Yeonsedae-gil, Wonju 220-710, Korea.

Purpose: Most chemical transfection reagents are ineffective for the transfection of cells in suspension, such as leukemic cell and stem cell lineages. We developed two different types of viroplexes, cationic Sendai F/HN viroplexes (CSVs) and protamine sulfate-condensed cationic Sendai F/HN viroplexes (PCSVs) for the efficient transfection of T-leukemic cells.

Materials And Methods: The viroplex systems were prepared by reconstitution of fusogenic Sendai F/HN proteins in DMKE (O,O'-dimyristyl-N-lysyl glutamate) cationic liposomes. The viroplexes were further optimized for plasmid DNA and siRNA delivery to suspension cells. The particle size and surface charge of the viroplexes were analyzed with a ζ-sizer. Transfection of plasmid DNA (pDNA) and small interfering RNA (siRNA) by CSVs or PCSV was evaluated by measurement of transgene expression, confocal microscopy, FACS, and RT-PCR.

Results: The optimized CSVs and PCSVs exhibited enhanced gene and siRNA delivery in the tested suspension cell lines (Jurkat cells and CEM cells), compared with conventional cationic liposomes. In the case of pDNA transfection, the CSVs and PCSVs show at least 10-fold and 100-fold higher transgene expression compared with DMKE lipoplexes (or lipofectamine 2000), respectively. The CSVs showed more effective siRNA delivery to the suspension cells than cationic liposomes, as assessed by confocal microscopy, FACS, and RT-PCR. The effective transfection by the CSVs and PCSVs is presumably due to fusogenic activity of F/HN proteins resulting in facilitated internalization of pDNA and siRNA.

Conclusion: This study suggests that Sendai F/HN viroplexes can be widely applicable for the transfection of pDNA and siRNA to suspension cell lines.
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http://dx.doi.org/10.3349/ymj.2013.54.5.1149DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3743179PMC
September 2013

The effect of ossicle resection in the lateral ligament repair for treatment of chronic lateral ankle instability.

Foot Ankle Int 2013 Aug 7;34(8):1128-33. Epub 2013 Mar 7.

Department of Orthopedic Surgery, Sungkyunkwan University School of Medicine, Seoul, Korea.

Background: The management of an ossicle or avulsion fragment of the fibular tip in chronic lateral ankle instability is an open question. Some authors maintain the necessity of osteosynthesis for reconstruction of the lateral ligamentous structure if the fragment is large. We hypothesized that the modified Broström procedure with resection of the ossicle would result in good outcomes compared to that of the same procedure for chronic lateral instability patients without ossicle.

Methods: Between December 2004 and December 2010, 102 patients underwent the modified Broström procedure for chronic lateral instability. Of these, 82 patients (86 ankles) were available for this study. Forty ankles had ossicles at the fibular tip (group O), 46 had no ossicle (group N). The average follow-up period was 33 months in group O and 37 months in group N. Irrespective of size, if there were ossicles we resected all fragments and performed the modified Broström procedure. To analyze the surgical outcome, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot pain and function scales and Karlsson scores were compared between the 2 groups preoperatively and postoperatively.

Results: Preoperative scores in the 2 groups showed no significant difference, except for AOFAS pain score. There was no significant difference in postoperative AOFAS pain and function score between the groups. Postoperative Karlsson score was significantly higher in group O than in group N (P = .001). Group O was divided into 2 subgroups by the largest diameter of the ossicle (< 10 mm and ≥ 10 mm); there was no significant difference in surgical outcomes.

Conclusions: In the treatment of chronic lateral instability of ankle, if there are ossicles on the fibular tip, osteosynthesis of the ossicles may not be necessary, even if the size is considerable. Modified Broström procedure after resection of the ossicle was successful.

Level Of Evidence: Level III, retrospective case series.
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http://dx.doi.org/10.1177/1071100713481457DOI Listing
August 2013

Comparison of cell-labeling methods with ¹²⁴I-FIAU and ⁶⁴Cu-PTSM for cell tracking using chronic myelogenous leukemia cells expressing HSV1-tk and firefly luciferase.

Cancer Biother Radiopharm 2012 Dec 25;27(10):719-28. Epub 2012 Sep 25.

Molecular Imaging Research Center, Korea Institute of Radiological and Medical Sciences-KIRAMS, Seoul, Republic of Korea.

Cell-tracking methods with molecular-imaging modality can monitor the biodistribution of cells. In this study, the direct-labeling method with ⁶⁴Cu-pyruvaldehyde-bis(N4-methylthiosemicarbazone) (⁶⁴Cu-PTSM), indirect cell-labeling methods with herpes simplex virus type 1-thymidine kinase (HSV1-tk)-mediated ¹²⁴I-2'-fluoro-2'-deoxy-1-β-D-arabinofuranosyl-5-iodouracil (¹²⁴I-FIAU) were comparatively investigated in vitro and in vivo for tracking of human chronic myelogenous leukemia cells. K562-TL was established by retroviral transduction of the HSV1-tk and firefly luciferase gene in the K562 cell. K562-TL cells were labeled with ⁶⁴Cu-PTSM or ¹²⁴I-FIAU. Cell labeling efficiency, viability, and radiolabels retention were compared in vitro. The biodistribution of radiolabeled K562-TL cells with each radiolabel and small-animal positron emission tomography imaging were performed. Additionally, in vivo and ex vivo bioluminescence imaging (BLI) and tissue reverse transcriptase-polymerase chain reaction (RT-PCR) analysis were used for confirming those results. K562-TL cells were efficiently labeled with both radiolabels. The radiolabel retention (%) of ¹²⁴I-FIAU (95.2%±1.1%) was fourfold higher than ⁶⁴Cu-PTSM (23.6%±0.7%) at 24 hours postlabeling. Viability of radiolabeled cells was statistically nonsignificant between ¹²⁴I-FIAU and ⁶⁴Cu-PTSM. The radioactivity of each radiolabeled cells was predominantly accumulated in the lungs and liver at 2 hours. Both the radioactivity of ⁶⁴Cu-PTSM- and ¹²⁴I-FIAU-labeled cells was highly accumulated in the liver at 24 hours. However, the radioactivity of ¹²⁴I-FIAU-labeled cells was markedly decreased from the body at 24 hours. The K562-TL cells were dominantly localized in the lungs and liver, which also verified by BLI and RT-PCR analysis at 2 and 24 hours postinjection. The ⁶⁴Cu-PTSM-labeled cell-tracking method is more efficient than ¹²⁴I-FIAU-labeled cell tracking, because of markedly decrease of radioactivity and fast efflux of ¹²⁴I-FIAU in vivo. In spite of a high labeling yield and radiolabel retention of ¹²⁴I-FIAU in vitro, the in vivo cell-tracking method using ⁶⁴Cu-PTSM could be a useful method to evaluate the distribution and targeting of various cell types, especially, stem cells and immune cells.
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http://dx.doi.org/10.1089/cbr.2012.1225DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3516418PMC
December 2012

Surgical reconstruction of chronic achilles tendon ruptures using various methods.

Orthopedics 2012 Feb 17;35(2):e213-8. Epub 2012 Feb 17.

Department of Orthopedic Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.

The purpose of this study was to evaluate the surgical outcomes of reconstruction of chronic Achilles tendon ruptures using various methods, including Achilles tendon allograft. Between October 2003 and March 2010, twelve patients with chronic Achilles tendon ruptures and a defect gap of >4 cm underwent surgical reconstruction with V-Y advancement, gastrocnemius fascial turn-down flap, flexor hallucis longus tendon transfer, or Achilles tendon allograft. The study group comprised 11 men and 1 woman. At last follow-up, all patients were assessed with regard to postoperative complications, self-reported level of satisfaction, American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot score, Achilles tendon Total Rupture Score, repetitive single-heel rises, single-leg hopping test, and ankle range of motion. The AOFAS scores increased from an average of 68.7 (range, 50-87) preoperatively to 98.0 (range, 88-100) postoperatively. All patients were able to perform 10 repetitive single-heel raises and single-leg hops at last follow-up. No patient experienced wound complications or deep infection. Seven patients were rated as excellent, 4 as good, and 1 as fair. Chronic Achilles tendon ruptures can be successfully treated by careful selection of the reconstruction method according to the length of defect gap and state of the remaining tissue. With an extensive defect, use of an Achilles tendon allograft can be a good option.
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http://dx.doi.org/10.3928/01477447-20120123-13DOI Listing
February 2012

Hepatic control elements promote long-term expression of human coagulation factor IX gene in hydrodynamically transfected mice.

J Gene Med 2011 Jul;13(7-8):365-72

Department of Biomedical Laboratory Science, Korea Nazarene University, Cheonan.

Background: Long-term expression of the delivered target gene is critical for successful gene therapy. Recently, hepatic control region I (HCR I) originating from the apolipoprotein (apo)C-I pseudogene was shown to be a critical element for long-term gene expression in the liver of mice. HCR II is another hepatic control region of apoC-I.

Methods: HCR I, HCR II and HCR I/II-containing plasmids encoding factor IX were prepared and hydrodynamically transferred into the liver of normal and hemophilia B mice. Factor IX expression, clotting activity and formation of antibodies against the expressed gene product were compared.

Results: HCR I-, HCR II- and HCR I/II-containing plasmids all induced long-term gene expression in both normal and hemophilia B mice. Post-transfection factor IX expression in the hemophilia B mice remained above 500 ng/ml for 210 days. Antibodies against human factor IX were detected at a low level in the serum, although they had no effect on the levels and clotting activity of the expressed factor IX.

Conclusions: We have shown in mouse models that hydrodynamic transfection of pBS-HCRII-HP-FIXA and pBS-HCRI/II-HP-FIXA was able to induce and maintain the expression and clotting activity of human factor IX for a long period of time at a potentially therapeutic level. With an appropriate delivery system, this type of plasmid vector could be clinically useful for the hepatic expression of therapeutic genes including human factor IX.
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http://dx.doi.org/10.1002/jgm.1583DOI Listing
July 2011

A Broken Drill-bit Fragment Causing Severe Radiating Pain after Cervical Total Disc Replacement: A Case Report.

Asian Spine J 2011 Jun 2;5(2):125-9. Epub 2011 May 2.

Department of Orthopaedic Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.

This is a case report of a 38-year-old man with severe radiating pain on upper extremity after cervical total disc replacement (TDR). We faced an unusual complication that has not been reported yet. He underwent cervical TDR for left central disc protrusion on C5-6. After the surgery, preoperative symptom disappeared. However, at postoperative 1 year, he complained severe right-sided radiating pain that had a sudden onset. On postoperative X-ray, a metal fragment which seemed like a broken drill bit was shown within the spinal canal. To remove that, right-sided anterior microforaminotomy on C5-6 was performed and the metal fragment was removed successfully. After that, anterior fusion was done because the motion of the artificial disc was minimal and the removed structure seemed to attenuate stability during cervical motion. The operation resulted in prompt symptomatic relief. During cervical TDR, particular attention should be paid to the procedures that require using drill-bits.
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http://dx.doi.org/10.4184/asj.2011.5.2.125DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3095802PMC
June 2011

Changes of upper thoracic curve and shoulder balance in thoracic adolescent idiopathic scoliosis treated by anterior selective thoracic fusion using VATS.

J Spinal Disord Tech 2011 Oct;24(7):462-8

Department of Orthopedic Surgery, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.

Study Design: A retrospective radiographic analysis.

Objectives: To evaluate changes of upper thoracic curve and shoulder balance in thoracic adolescent idiopathic scoliosis patients treated by anterior selective thoracic fusion using video-assisted thoracoscopic surgery and to identify adequacy of earlier criteria of double thoracic (DT) curve for anterior correction.

Summary Of Background Data: Although anterior and posterior scoliosis correction show many differences in correction mechanisms, fusion levels, loss of correction etc., the criteria of DT curve was applied without differences. There are no reports about these differences.

Materials And Methods: Forty patients were followed for a minimum of 3 years (range, 3-8 y). The magnitude and flexibility of upper thoracic, lower thoracic, and the superior portion of the lower thoracic curve were measured using full length standing and side-bending radiographs before surgery, at 1 week postoperatively, and at last follow-up. The correction rate and loss of correction of these curves were calculated and preoperative and postoperative radiographic shoulder heights (RSHs) were measured. RSH was defined as balanced (shoulder height difference <10 mm), mildly imbalanced (10-20 mm), or moderately imbalanced (>20 mm). T1 tilt and coronal balance were also evaluated. Patients were divided into groups based on these factors and postoperative RSH was compared.

Results: Flexibility of the upper thoracic curve was 46% and magnitude of the upper thoracic curve was corrected spontaneously from 28.6±7.8 degrees to 17.9±7.0 degrees with a 37.4% correction rate that did not change during follow-up. On average, preoperative left shoulder was 6.3±10.5 mm lower than right shoulder and this changed to 10.4±11.8 mm and 6.0±8.2 mm higher than right shoulder at 1 week postoperatively and at last follow-up, respectively. The group with an upper thoracic curve of ≥30 degrees or a superior portion of the lower thoracic curve of ≥30 degrees preoperatively had a higher left shoulder postoperatively (P=0.016, 0.040). Of the 12 patients with a symmetric or higher left shoulder (≥0 mm) preoperatively, 9 patients had a balanced shoulder (-10-10 mm) and 3 patients showed mild shoulder imbalance (<20 mm) at last follow-up.

Conclusions: Among patients who have DT curve, patients with mild left shoulder elevation (<20 mm) can be treated by anterior correction unless the magnitude of upper thoracic curve or superior portion of lower thoracic curve are ≥30 degrees. For anterior correction, criteria of DT curve might be applied less strictly.
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http://dx.doi.org/10.1097/BSD.0b013e318204d553DOI Listing
October 2011

Arthroscopic treatment of popliteal cysts: clinical and magnetic resonance imaging results.

Arthroscopy 2010 Oct 24;26(10):1340-7. Epub 2010 Sep 24.

Department of Orthopaedic Surgery, Sungkyunkwan University School of Medicine, Samsung Medical Center, Seoul, South Korea.

Purpose: This study examined the functional and magnetic resonance imaging (MRI) outcomes of popliteal cysts with combined intra-articular pathologies that were treated arthroscopically by decompression and a cystectomy through an additional posteromedial cystic portal.

Methods: From January 2003 to March 2008, 31 patients were treated with a modified arthroscopic technique to decompress a popliteal cyst. The connecting valvular mechanism was found in all cases at the posteromedial compartment through the anterolateral viewing portal, and it was corrected by resecting the capsular fold through the posteromedial working portal. For cysts with multiple fibrous septa, an additional portal, the so-called posteromedial cystic portal, was used for complete cyst removal. The functional outcome was evaluated by use of the Rauschning and Lindgren knee score. All patients were evaluated by MRI, which documented the popliteal cyst and associated intra-articular lesions preoperatively and at follow-up.

Results: All patients could return to their previous daily activities with few or no limitations, and no additional surgery was required after a mean follow-up of 36.1 months (range, 12 to 72 months). The Rauschning and Lindgren knee score showed improved clinical features at the final follow-up in 94% of patients. The follow-up MRI study showed that the cyst had disappeared in 17 knees (55%) and had reduced in size in 14 knees (45%) in the 31 patients. The mean cyst size was reduced significantly from 6.8 to 0.8 cm (P < .0001).

Conclusions: The described arthroscopic technique with or without an additional posteromedial cystic portal is effective for treating popliteal cysts with combined intra-articular lesions. More importantly, follow-up MRI showed that the cyst size was reduced or it had disappeared in all cases, although there was no association between the cyst's disappearance and the follow-up clinical score.

Level Of Evidence: Level IV, therapeutic case series.
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http://dx.doi.org/10.1016/j.arthro.2010.02.012DOI Listing
October 2010

Leukemia-specific siRNA delivery by immunonanoplexes consisting of anti-JL1 minibody conjugated to oligo-9 Arg-peptides.

Mol Cells 2010 May 12;29(5):457-62. Epub 2010 Apr 12.

Department of Biomedical Laboratory Science, Yonsei University, Wonju, 220-710, Korea.

Targeted mRNA degradation by short interfering RNAs (siRNAs) offers a great potential to treat cancers. siRNA therapeutics for leukemias are, however, hindered by poor intracellular uptake, limited blood stability and nonspecific delivery. To solve these problems, we developed an anti-JL1 immunonanoplex (antibody-coupled nanocomplex) for siRNA delivery using anti-JL1 minibody (leukemia cell-specific minibody) conjugated to oligo-9-Arg peptide (9R) for effective siRNA delivery to leukemic cells. The anti-JL1 immunonanoplexes were able to deliver siRNA specifically to leukemic cells (CEM and Jurkat), but not to control cancer cells (H9). According to FACS and confocal microscopic analysis, siRNAs delivered by immunonanoplex particles were rapidly taken up by the JL1-positive cancer cells in 2 h. Furthermore, we showed that the anti-JL1 immunonanoplexes were effectively targeted to JL1-positive cells (CEM) inoculated in the mouse bone marrow. These results suggest that the anti-JL1 immunonanoplex is a powerful siRNA delivery system for human leukemia therapies.
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http://dx.doi.org/10.1007/s10059-010-0056-5DOI Listing
May 2010

Non-operative treatment of ACL rupture with mild instability.

Arch Orthop Trauma Surg 2010 Aug 25;130(8):1001-6. Epub 2010 Mar 25.

Department of Orthopaedic Surgery, Samsung Medical Center, Sungkyunkwan University, Kangnam-Ku, Seoul, Korea.

Purpose: This study examined whether or not conservative treatment of an acutely injured anterior cruciate ligament (ACL) could be successful in a select group of patients. Routine ACL reconstruction surgery in all acute ACL-injured patients should be avoided. We hypothesize that acutely injured ACL with mild instability at the initial physical examination could be improved even if there is disruption of ACL fibers on magnetic resonance images (MRI).

Materials And Methods: Among 232 acute ACL-injured patients who visited our institution from March 1997 to April 2006, 48 were treated non-operatively. Patients diagnosed with an acute ACL injury by MRI with Lachman test < or =grade 1 were treated non-operatively. In this study, 30 male and 18 female patients with a mean age of 31.8 years were enrolled. The initial and follow-up Lachman test and pivot shift test were performed 3 weeks after the injury. The Lysholm knee scoring scale, International Knee Documentation Committee (IKDC) score and KT-2000 were obtained at the last follow-up.

Results: There were 12 complete (25%) and 36 incomplete ACL ruptures (75%). The patients were followed up clinically and with MRI for 21.5 and 11.3 months, respectively. The follow-up Lachman test improved to grade 0 in 41 patients (87%). Thirty-six patients (76%) showed no laxity in the follow-up pivot shift test. The last follow-up IKDC score was a mean value of 91.1 points. KT 2000 was performed in 40 patients with a mean side-to-side difference of 2.85 mm. Of 48 patients, 46 showed restored ACL continuity and 39 (84%) showed restored low signal intensity on MRI.

Conclusion: A selective group of ACL tears with mild instability (Lachman < or =grade 1), though these seem to be complete tears on MRI, can show restoration of their continuity and signals on the MRI. Joint laxity on physical examination was improved at follow-up. These results suggest that a select group of patients with an acute ACL injury can successfully undergo non-operative treatment. In addition, unnecessary early ACL reconstruction surgery should be avoided.
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http://dx.doi.org/10.1007/s00402-010-1077-4DOI Listing
August 2010

Correlation between dynamic postural stability and muscle strength, anterior instability, and knee scale in anterior cruciate ligament deficient knees.

Arch Orthop Trauma Surg 2010 Aug 9;130(8):1013-8. Epub 2010 Mar 9.

Department of Physical Medicine and Rehabilitation, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, Korea.

Aim: The purpose of this study was to assess the correlations between dynamic postural stability and muscle strength, anterior instability, and knee scores in anterior cruciate ligament (ACL) deficient knees.

Method: We examined 40 male patients with ACL injury prior to surgery. Maximal torques of flexors and extensors of the injured knee at 60 and 180 degrees /s were evaluated using an isokinetic testing device. Anterior translations of the tibia were measured using a KT-2000 arthrometer, and dynamic postural stabilities were measured during single-leg stance using the Biodex Stability System (BSS). Knee statuses were evaluated using Lysholm and International Knee Documentation Committee (IKDC) knee scores. Correlations between dynamic postural stability and muscle strength, anterior instability, and knee scores were determined.

Results: Significant correlations were found between maximal torques of the extensor of injured knees at 60 and 180 degrees /s (r = -0.52, p = 0.048 and r = -0.46, p = 0.019, respectively) and dynamic postural stability, and maximal torques of flexors of injured knees showed similar relations (r = -0.51, p = 0.0048 and r = -0.47, p = 0.016, respectively). Lysholm and IKDC knee scores were also found to be correlated with dynamic postural stability (r = -0.49, p = 0.001 and r = -0.52, p = 0.005, respectively). However, no correlation was found between grade of anterior translation measured using the KT-2000 arthrometer and dynamic postural stability (p = 1.0).

Conclusion: Dynamic postural stability determined using the BSS appears to be influenced by muscle strength, as determined by isokinetic testing, but not with grade of anterior translation measured using a KT-2000 arthrometer. Subjective knee scores appear to improve in parallel with dynamic postural stability.
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http://dx.doi.org/10.1007/s00402-010-1080-9DOI Listing
August 2010

In vitro and in vivo gene-transferring characteristics of novel cationic lipids, DMKD (O,O'-dimyristyl-N-lysyl aspartate) and DMKE (O,O'-dimyristyl-N-lysyl glutamate).

J Control Release 2006 Oct 11;115(2):234-41. Epub 2006 Aug 11.

Department of Biomedical Laboratory Science, Yonsei University, Wonju 220-710, Republic of Korea.

We chemically synthesized two different cationic lipids consisting of a core of lysine, two C-14 hydrocarbon chains, and either aspartatic acid or glutamic acid. The lipids were assigned the acronyms, DMKD and DMKE. Cationic liposomes prepared with the two different lipids were tested for their gene-transferring capabilities in various cell lines compared with that of control DOTAP liposomes. Under the same experimental conditions, the order of in vitro gene transfection efficiency was DMKE>or=DMKD>DOTAP. To identify the parameters influencing transfection efficiency, the DNA-binding affinities of the liposomes were compared and changes in particle size and surface charge were examined after complex formation. Both the DNA-binding affinity of the liposomes and the cell surface-binding affinity of the liposome-pDNA complexes were crucial for gene transfection. In addition, intravenously administered DMKE and DMKD liposomes exhibited different biodistribution characteristics and intensity of in vivo organ transfection from the DOTAP liposomes. Compared to the DOTAP liposomes, they were more readily transferred to the liver. Interestingly, when they were directly injected into tumor tissues, the DMKE lipoplexes were able to induce more efficient transgene expression in these tissues than the DOTAP and DMKD lipoplexes. This study suggests that a small difference in the cationic lipid backbone structure significantly affects gene-transferring capabilities. DMKE and DMKD liposomes can be utilized as efficient gene-transferring vehicles for hepatic or intra-tumoral gene transfection.
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http://dx.doi.org/10.1016/j.jconrel.2006.08.003DOI Listing
October 2006

Antitumor effects of angiostatin K1-3 and endostatin genes coadministered by the hydrodynamics-based transfection method.

Oncol Res 2005 ;15(7-8):343-50

Department of Biomedical Laboratory Science, College of Health Science, Yonsei University, Wonju 220-710, Republic of Korea.

Angiostatin and endostatin are potent endothelial cell growth inhibitors and have been carefully evaluated for antiangiogenic cancer therapy. Previously, we have shown that subcutaneous administration of angiostatin K1-3 and endostatin genes complexed with liposomal vectors is a more practical treatment procedure than administration of angiostatin and endostatin proteins. This study provides additional conclusive evidence supporting the effectiveness of antiangiogenic cancer gene therapy employing angiostatin K1-3 and endostatin genes. Plasmids encoding a mouse angiostatin K1-3 gene (pFLAG-AngioK1/3) and an endostatin gene (pFLAG-Endo) were introduced by the hydrodynamic transduction method into mice carrying Matrigel plugs or B16BL6 mouse melanoma tumors. A single systemic injection of the two genes exhibited potent antiangiogenic and antitumor activity in the mouse model. Hydrodynamic coadministration of the genes inhibited the B16BL6 mouse melanoma growth and pulmonary metastasis more effectively than administration of either gene alone. Compared with the untreated control group, the mice cotreated with pFLAG-AngioK1/3 and pFLAG-Endo exhibited 75% reduction of tumor growth while those treated with pFLAG-AngioK1/3 or pFLAG-Endo showed 46% and 52% reduction, respectively. The cotreatment inhibited B16BL6 pulmonary metastasis formation by 80% while the inhibition induced by individual treatment with pFLAG-AngioK1/3 or pFLAG-Endo was 68% and 71%, respectively. These results provide additional evidence that systemic expression of angiostatin K1-3 and/or endostatin genes is a viable alternative procedure for antiangiogenic cancer therapy.
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http://dx.doi.org/10.3727/096504005776449707DOI Listing
March 2006

Gene-transferring efficiencies of novel diamino cationic lipids with varied hydrocarbon chains.

Bioconjug Chem 2004 Sep-Oct;15(5):1095-101

Department of Biomedical Laboratory Science, Yonsei University, Wonju 220-710, Korea.

Utilizing three biocompatible components, a series of novel cationic lipids has been chemically synthesized and tested for their gene-transferring capabilities in 293 transformed kidney cells and B16BL6 mouse melanoma cells. The synthesized cationic lipids consisting of a core of lysine and aspartic acid with hydrocarbon chains of varied length were assigned the acronyms DLKD (O,O'-dilauryl N-lysylaspartate), DMKD (O,O'-dimyristyl N-lysylaspartate), DPKD (O,O'-dipalmityl N-lysylaspartate), and DSKD (O,O'-distearyl N-lysylaspartate). The gene-transferring capabilities of these cationic lipids were found to be dependent on the hydrocarbon chain length. Under similar experimental conditions, the order of gene transfection efficiency was DMKD > DLKD > DPKD > DSKD. Addition of cholesterol or dioleoyl phosphatidylethanolamine (DOPE) as a colipid did not change this order. Colipid addition affected the transfection efficiency positively or negatively depending on the length of the cationic lipid acyl chain. On the whole, the length of the hydrophobic carbon chain was a major factor governing the gene-transferring capabilities of this series of cationic lipids. The observed differences in transfection efficiency may be due to differing binding affinities to DNA molecules as well as differences in the surface charge potential of the liposome-DNA complexes (lipoplexes) in the aqueous environment.
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http://dx.doi.org/10.1021/bc049934tDOI Listing
February 2005

Inhibition of B16BL6 tumor progression by coadministration of recombinant angiostatin K1-3 and endostatin genes with cationic liposomes.

Cancer Gene Ther 2004 Jun;11(6):441-9

Department of Biomedical Laboratory Science and Institute of Health Science, Yonsei University, Wonju 220-710, Republic of Korea.

Transfection of the antiangiogenic angiostatin and endostatin genes was shown to be an alternative to high-dose administration of angiostatin or endostatin proteins for cancer therapy. We have systematically investigated whether coadministration of the mouse angiostatin kringle 1-3 gene (pFLAG-AngioK1/3) and the endostatin gene (pFLAG-Endo) complexed with cationic liposomes exhibits enhanced therapeutic efficacy. In vitro, the coexpressed mixture of angiostatin K1-3 and endostatin more effectively reduced angiogenesis in chorioallantoic membranes than either angiostatin K1-3 or endostatin alone. In vivo, subcutaneous co-administration of pFLAG-AngioK1/3 and pFLAG-Endo lipoplexes more effectively inhibited vascularization in Matrigel plugs implanted in mice than either one alone. Additionally, subcutaneous administration of these genes inhibited the growth and formation of pulmonary metastases of B16BL6 melanoma cells in mice. Compared to treatment with an empty vector, treatment with pFLAG-AngioK1/3 plus pFLAG-Endo inhibited 81% of tumor growth, while treatment with pFLAG-AngioK1/3 or pFLAG-Endo inhibited tumor growth 70 and 69%, respectively. Cotreatment with the two plasmids after primary tumor excision induced a 90% inhibition of pulmonary metastases versus 79% for pFLAG-AngioK1/3 or 80% for pFLAG-Endo individually. These results suggest that combined administration of angiostatin K1-3 and endostatin genes complexed with cationic liposomes may be an innovated antiangiogenic strategy for cancer therapy.
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http://dx.doi.org/10.1038/sj.cgt.7700716DOI Listing
June 2004

Inhibition of angiogenesis by salmosin expressed in vitro.

Oncol Res 2004 ;14(4-5):227-33

Department of Biomedical Laboratory Science and Institute of Health Science, Yonsei University, Wonju 220-710, Republic of Korea.

Recently, salmosin, a novel snake venom-derived disintegrin containing the Arg-Gly-Asp (RGD) sequence, was reported to be both antiangiogenic and antitumorigenic. The antitumor activity was substantiated by in vivo administration of recombinant salmosin into mice bearing tumors. However, it was difficult to prepare functionally active recombinant salmosin and to maintain a therapeutically effective concentration of the protein in the circulatory system by daily injections. Hence, we have suggested that salmosin gene transfer mediated by cationic liposomes may be a practical alternative for cancer treatment. Plasmids encoding the salmosin gene were constructed and then transferred by means of cationic liposomes into transformed human embryonic kidney (HEK) 293 cells. The transfected genes were able to produce functionally active salmosin proteins in vitro. In fact, the expressed salmosin remarkably inhibited proliferation of bovine capillary endothelial (BCE) cells and effectively inhibited the migration of highly metastatic B16BL6 mouse melanoma cells. Neovascularization in chick chorio-allantoic membranes (CAM) and in Matrigel implanted subcutaneously into mice was greatly inhibited in the presence of the expressed salmosin. Based on these experimental results, we suggest that the antitumor effect induced by salmosin gene transfection may be due to the antiangiogenic activity of the expressed salmosin proteins.
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http://dx.doi.org/10.3727/000000003772505353DOI Listing
September 2004

Inhibitory effect of the salmosin gene transferred by cationic liposomes on the progression of B16BL6 tumors.

Cancer Res 2003 Oct;63(19):6458-62

Department of Biomedical Laboratory Science and Institute of Health Science, Yonsei University, Wonju 220-710, South Korea.

Salmosin is a novel disintegrin containing the Arg-Gly-Asp sequence that significantly inhibits platelet aggregation, basic fibroblast growth factor-induced endothelial cell proliferation, and tumor progression by antagonizing integrin-mediated cell interactions. Previously, it was shown that daily administration of salmosin was able to inhibit tumor-derived angiogenesis and adherence and proliferation of tumor cells, resulting in suppression of tumor progression. However, it is very difficult to maintain a therapeutic level of salmosin in the blood by systemic administration of the protein. Hence, an alternative strategy for antiangiogenic cancer therapy, based on the in vivo expression of the salmosin gene administered with cationic liposomes, was investigated. The salmosin peptides expressed in vitro inhibited the proliferation of bovine capillary endothelial cells in a dose-dependent manner, presumably as a result of inhibition of cell adhesion mediated via alpha(v)beta(3) integrin. Subcutaneous administration of the salmosin gene resulted in systemic expression of the gene product and concomitant inhibition of the growth of B16BL6 melanoma cells. Suppression of pulmonary metastases, verified by experimental and spontaneous metastasis models in mice, also resulted from salmosin gene treatment. These results suggest that administration of the salmosin gene complexed to cationic liposomes is effective in maintaining antiangiogenic salmosin at an effective therapeutic level and may be clinically applicable to anticancer gene therapy.
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October 2003