Low Molecular Weight Protamine/ Insulin Formulation with Potential to Attenuate Protamine-Masqueraded Insulin Allergy

Huining He,Victor C. Yang,Allan E. David,Jian Zhang,박윤신,Jianxin Wang,Yongzhuo Huang,Jingkang Wang 한국고분자학회 2011 Macromolecular Research Vol.19 No.12

Construction and characterization of gelonin and saporin plasmids for toxic gene-based cancer therapy

Min, Kyoung Ah,He, Huining,Yang, Victor C.,Shin, Meong Cheol Springer-Verlag 2016 Archives of Pharmacal Research Vol.39 No.5

<P>Toxic gene therapy (or suicidal gene therapy) is gaining enormous interest, specifically for the treatment of cancer. The success of this therapy lies in several crucial factors, including the potency of gene products to kill the transfected tumor cells and the transfection ability of the transfection vehicles. To address the potency problem, in the present study, we engineered two separate mammalian transfection plasmids (pSAP and pGEL) containing genes encoding ribosome inactivating proteins (RIPs), gelonin and saporin. After the successful preparation and amplification of the plasmids, they were tested on various cancer cell lines (HeLa, U87, 9L, and MDA-MB-435) and a noncancerous cell line (293 HEK) using polyethyleneimine (PEI) as the transfection agent. Transfection studies performed under varying gene concentration, incubation time, and gene-to-PEI ratios revealed that, compared to the treatment of pGFP (GFP expression plasmid)/PEI, both pGEL/PEI and pSAP/PEI complexes could induce significantly augmented cytotoxic effects at only 2 mu g/mL gene concentration. Importantly, these cytotoxic effects were observed universally in all tested cancer cell lines. Overall, this study demonstrated the potential of pGEL and pSAP as effective gene candidates for the toxic gene-based cancer therapy.</P>

Tandem-multimeric F3-gelonin fusion toxins for enhanced anti-cancer activity for prostate cancer treatment

Shin, Meong Cheol,Min, Kyoung Ah,Cheong, Heesun,Moon, Cheol,Huang, Yongzhuo,He, Huining,Yang, Victor C. Elsevier/North Holland 2017 International journal of pharmaceutics Vol.524 No.1

<P><B>Abstract</B></P> <P>Despite significant progress in prostate cancer treatment, yet, it remains the leading diagnosed cancer and is responsible for high incidence of cancer related deaths in the U.S. Because of the insufficient efficacy of small molecule anti-cancer drugs, significant interest has been drawn to more potent macromolecular agents such as gelonin, a plant-derived ribosome inactivating protein (RIP) that efficiently inhibits protein translation. However, in spite of the great potency to kill tumor cells, gelonin lacks ability to internalize tumor cells and furthermore, cannot distinguish between tumor and normal cells. To address this challenge, we genetically engineered gelonin fusion proteins with varied numbers of F3 peptide possessing homing ability to various cancer cells and angiogenic blood vessels. The <I>E. coli</I> produced F3-gelonin fusion proteins possessed equipotent activity to inhibit protein translation in cell-free protein translation systems to unmodified gelonin; however, they displayed higher cell uptake that led to significantly augmented cytotoxicity. Compared with gelonin fusion with one F3 peptide (F3-Gel), tandem-multimeric F3-gelonins showed even greater cell internalization and tumor cell killing ability. Moreover, when tested against LNCaP <I>s.c.</I> xenograft tumor bearing mice, more significant tumor growth inhibition was observed from the mice treated with tandem-multimeric F3-gelonins. Overall, this research demonstrated the potential of utilizing tandem multimeric F3-modified gelonin as highly effective anticancer agents to overcome the limitations of current chemotherapeutic drugs.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Cellular uptake mechanism and comparative in vitro cytotoxicity studies of monomeric LMWP-siRNA conjugate

Junxiao Ye,Xing Pei,Hui Cui,Zhili Yu,이혁진,Jian-Xin Wang,Xu Wang,Lu Sun,Huining He,Victor C. Yang 한국공업화학회 2018 Journal of Industrial and Engineering Chemistry Vol.63 No.-

The covalent attachment of CPPs to siRNA molecules offers great potential for CPP-mediated siRNA delivery. We recently reported a concise and high-yield synthesis strategy of the cell-permeable, cytosol-dissociable LMWP-siRNA covalent conjugate. Herein, cell uptake mechanism and cellular toxicity studies of this conjugate were performed to evaluate the potential of LMWP-siRNA conjugate for clinical translation. Cellular uptake mechanism study indicated that the conjugate could be taken up by cells via multiple pathways, including direct penetration of the plasma membrane and clathrin- and caveolae-independent endocytosis. In vitro cytotoxicity study revealed that the conjugation promoted internalization in a low-toxic fashion.

Specific down regulation of 3T3-L1 adipocyte differentiation by cell-permeable antisense HIF1α-oligonucleotide

Park, Yoon Shin,Huang, Yongzhuo,Park, Yoon Jeong,David, Allan E.,White, Lindsay,He, Huining,Chung, Hee Sun,Yang, Victor C. Elsevier 2010 Journal of controlled release Vol.144 No.1

<P><B>Abstract</B></P><P>Hypoxia is a strong modulator of angiogenesis, accelerating adipose tissue expansion, suggesting that hypoxia inducible factor 1α (HIF1α) can be a novel target for anti-obesity. We conjugated antisense-HIF1α-oligonucleotide (ASO) with low molecular weight protamine (LMWP), a cell-penetrating peptide, to enhance its ability to block hypoxic-angiogenesis, thereby eliciting an anti-obesity effect. Nano-sized ASO-LMWP (AS-L) conjugates enhanced cellular uptake of ASO without yielding a cytotoxic effect and protected the ASO against enzymatic attack and chemical reduction. AS-L showed enhanced intra-cellular localization compared to naked ASO and the complex of ASO with lipofectamine during hypoxic-differentiation. Consequently AS-L induced significant down-regulation of leptin and VEGF gene expressions, thereby reducing fat accumulation in the cell.</P><P>This proof-of-concept study shows that AS-L produces an inhibitory effect on adipogenesis and angiogenesis during differentiation, indicating LMWP mediated ASO delivery can potentially be a safe and promising treatment for obesity.</P> <P><B>Graphical abstract</B></P><P></P>

MMP-2-responsive fluorescent nanoprobes for enhanced selectivity of tumor cell uptake and imaging

Sun, Lu,Xie, Shuping,Ji, Xiuru,Zhang, Jingming,Wang, Dongmei,Lee, Seung Jin,Lee, Hyukjin,He, Huining,Yang, Victor C. The Royal Society of Chemistry 2018 Biomaterials Science Vol.6 No.10

<P>It is difficult to develop highly selective substrate-based fluorescent nanoprobes for specific matrix metalloproteinases (MMPs) due to overlapping substrate specificities among the family of MMP enzymes. To resolve this issue, we have developed novel fluorescent nanoprobes that are highly selective for soluble MMP-2. Herein, MMP-2-responsive nanoprobes were prepared by immobilizing fluorescent fusion proteins on nickel ferrite nanoparticles <I>via</I> the His-tag nickel chelation mechanism. The fusion protein consisted of a fluorescent mCherry protein with a cell penetrating peptide (CPP) moiety. An MMP-2 cleavage site was also introduced within the fusion protein, which was directly linked to the nickel ferrite nanoparticles. The selectivity of nanoprobes was modulated by hiding the cleavage site of MMP-2 substrates deeply inside the system, which could result in strong steric hindrance between the nanoprobes and MMPs, especially for membrane-tethered MMPs such as MMP-14. A cell-based assay demonstrated that the nanoprobes could only be activated by tumor cells secreting soluble MMP-2, but not membrane-tethered MMP-14. To further evaluate the contribution of the steric hindrance effect on the nanoprobes, a truncated recombinant MMP-14 was employed to confer their cleavage activity as compared to native membrane-tethered MMP-14. Furthermore, a designed probe with a diminished steric hindrance effect was proved to be activated by membrane-tethered type MMP-14. The results indicated that the design of fluorescent nanoprobes employing the steric hindrance effect can greatly enhance the selectivity of MMP-responsive nanoprobes realizing the specific detection of soluble MMP-2 in a tumor microenvironment. We believe that highly selective MMP-2-responsive fluorescent nanoprobes have broad impacts on biomedical applications including molecular imaging and labeling for tumor detection.</P>

Cardiac-derived stem cell engineered with constitutively active HIF-1a gene enhances blood perfusion of hindlimb ischemia

Xing Pei,Jiyoung Shin,김희정,Nana Wang,Chaewon Seo,Miyun Yoon,Xiongwen Chen,Jianqing Gao,Victor C. Yang,Huining He,Seungjin Lee 한국공업화학회 2022 Journal of Industrial and Engineering Chemistry Vol.105 No.-

Stem cell-based therapeutic approach provides a possible treatment for critical limb ischemia (CLI) byinducing revascularization and regenerating ischemic tissue. However, the clinical benefit is modestdue to low cell survival and limited efficacy after transplantation. Cardiac-derived stem cells (CSCs) mightbe a novel cell source for CLI treatment owing to their superb endothelial differentiation potential andangiogenic paracrine functions. In this study, the angiogenic ability of CSCs was maximized by geneticengineering with constitutively active form of hypoxia-inducible factor-1a (CA-HIF-1a), resistant tooxygen-dependent degradation. CSCs transfected with CA-HIF-1a (CA-HIF-CSCs) promoted supplementaryexpression of proangiogenic factors including VEGF, bFGF, Ang-1 and PDGF-B, along with enhancedangiogenic function including migratory effect, tube formation and endothelial differentiation potential. In the mouse CLI model, CA-HIF-CSCs transplanted into the ischemic region using fibrin gel as cell deliveryvehicle, improved blood perfusion and limb functional recovery with minimal incidence of foot necrosisand limb loss by promoting new vessel formation. Histological evidence further confirmed that CAHIF-CSC/gel treatment markedly alleviated muscle degeneration and fibrosis. CSCs genetically engineeredwith constitutively active HIF-1a provide a novel therapeutic modality in CLI combining stem cell andgene therapy.

Cellular uptake mechanism and comparative <i>in vitro</i> cytotoxicity studies of monomeric LMWP-siRNA conjugate

Ye, Junxiao,Pei, Xing,Cui, Hui,Yu, Zhili,Lee, Hyukjin,Wang, Jianxin,Wang, Xu,Sun, Lu,He, Huining,Yang, Victor C. THE KOREAN SOCIETY OF INDUSTRIAL AND ENGINEERING 2018 JOURNAL OF INDUSTRIAL AND ENGINEERING CHEMISTRY -S Vol.63 No.-

<P><B>Abstract</B></P> <P>The covalent attachment of CPPs to siRNA molecules offers great potential for CPP-mediated siRNA delivery. We recently reported a concise and high-yield synthesis strategy of the cell-permeable, cytosol-dissociable LMWP-siRNA covalent conjugate. Herein, cell uptake mechanism and cellular toxicity studies of this conjugate were performed to evaluate the potential of LMWP-siRNA conjugate for clinical translation. Cellular uptake mechanism study indicated that the conjugate could be taken up by cells via multiple pathways, including direct penetration of the plasma membrane and clathrin- and caveolae-independent endocytosis. <I>In vitro</I> cytotoxicity study revealed that the conjugation promoted internalization in a low-toxic fashion.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>