Utilizing carbon dots as non-viral vectors in gene delivery

박소연,한지은,나건 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-

Vectors are the most important component for gene therapy. The viral vectors are widely used because of their higher transfection efficiency. However, viral vectors have limitation of genetic material size, oncogenic potential and immunogenicity. Thus, the non-viral vectors have been developing to substitute viral vectors. In this study we utilized carbon dots (c-dots) as a non-viral vector and c-dots have unique properties such as great water solubility and low toxicity. The c-dots were generated using cationic polymer for gene delivery. The therapeutic gene complexed with c-dots (gene/c-dots) were transfected into human mesenchymal stem cells (hMSCs). C-dots have been found to be safer because c-dots are less toxic than cationic polymer. In addition, c-dots enable bioimaging after transfection of gene/c-dots into hMSCs (gene/c-dots@hMSCs). Even, the amount of secreted therapeutic protein from gene/c-dots@hMSCs was 25-fold higher than that from gene/cationic polymer@hMSCs.

유전자치료를 위한 벡터 개발의 연구 동향 : A Current Research Insight

손은화,손은수,표석능 한국약제학회 2004 Journal of Pharmaceutical Investigation Vol.34 No.5

The basic concept underlying gene therapy is that human diseases may be treated by the transfer of genetics material into specific cells of a patient in order to correct or supplement defective genes responsible for disease development. There are several systems that can be used to transfer foreign genetic material into the human body. Both viral and non-viral vectors are developed and evaluated for delivering therapeutic genes. Viral vectors are biological systems derived from naturally evolved viruses capable of transferring their genetics materials into host cells. However, the limitations associated with viral vectors, in terms of their safety, particularly immunogenecity, and their limited capacity of transgenic materials, have encouraged researchers to increasingly focus on non-viral vectors as an alternative to viral vectors. Although non-viral vectors are less efficient than viral ones, they have the advantages of safety, simplicity of preparation and high gene encapsulation capability. This article reviews the most recent studies highlighting the advantages and the limitation of gene delivery systems focused on non-viral systems compared to viral systems.

Recent advances in the development of gene delivery systems

YK sung,SW KIM 한국생체재료학회 2019 생체재료학회지 Vol.23 No.2

Background: Gene delivery systems are essentially necessary for the gene therapy of human genetic diseases. Gene therapy is the unique way that is able to use the adjustable gene to cure any disease. The gene therapy is one of promising therapies for a number of diseases such as inherited disorders, viral infection and cancers. The useful results of gene delivery systems depend open the adjustable targeting gene delivery systems. Some of successful gene delivery systems have recently reported for the practical application of gene therapy. Main body: The recent developments of viral gene delivery systems and non-viral gene delivery systems for gene therapy have briefly reviewed. The viral gene delivery systems have discussed for the viral vectors based on DNA, RNA and oncolytic viral vectors. The non-viral gene delivery systems have also treated for the physicochemical approaches such as physical methods and chemical methods. Several kinds of successful gene delivery systems have briefly discussed on the bases of the gene delivery systems such as cationic polymers, poly(L-lysine), polysaccharides, and poly(ethylenimine)s. Conclusion: The goal of the research for gene delivery system is to develop the clinically relevant vectors such as viral and non-viral vectors that use to combat elusive diseases such as AIDS, cancer, Alzheimer, etc. Next step research will focus on advancing DNA and RNA molecular technologies to become the standard treatment options in the clinical area of biomedical application.

Therapeutic gene delivery of using stem cells as tumor specific non-viral vector and photochemical internalization

박소연,한지은,황희숙,나건 한국공업화학회 2019 한국공업화학회 연구논문 초록집 Vol.2019 No.0

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is promising therapeutic protein to induce tumor apoptosis. TRAIL gene is delivered by non-viral vector due to short half life and low stability. However, gene delivery using non-viral vector has limitation on serum stability and target specificity to tumor. To overcome the limitation of gene delivery using non-viral vector, human mesenchymal stem cells (hMSCs) and photochemical internalization (PCI) strategy were utilized for targeting tumor and enhanced transfection efficiency. We substantiated that homing ability of hMSCs could enhance tumor targeting and PCI-mediated TRAIL gene loading in hMSCs induced TRAIL gene expression at tumor site. Also, the xenograft mouse model was observed that TRAIL protein was released from hMSCs efficiently after condition of laser irradiation leading to a valuable therapeutic efficiency as an antitumor effect.

Human mesenchymal stem cell loading therapeutic gene by photo-mediated system

박소연,한지은,황희숙,나건 한국공업화학회 2019 한국공업화학회 연구논문 초록집 Vol.2019 No.1

Therapeutic gene indicates short half life and low stability due to denaturation. Therefore, the therapeutic gene is complexed with nonviral vectors such as cationic polymers or lipids. In this study, cationic polymer was used as non-viral vector, and complexed with therapeutic gene (polyplex). Non-viral vectors are safer than viral vectors however there are many obstacles that non-viral vectors face long delivery pathways. To overcome the limitation of gene therapy with non-viral vectors, we used human mesenchymal stem cells (hMSCs) to target tumor and photochemical internalization (PCI) to enhance transfection efficiency. We demonstrate that therapeutic gene loaded hMSCs (polyplex@hMSCs) facilitate tumor targeting and PCI could increase the expression of therapeutic gene in polyplex@hMSCs. All the taken together, PCI-mediated polyplex@hMSCs (PCI-polyplex@hMSCs) offer exciting potential therapeutic effect in cancer gene therapy.

Non-viral systemic delivery of Fas siRNA suppresses cyclophosphamide-induced diabetes in NOD mice

Jeong, J.H.,Kim, S.H.,Lee, M.,Kim, W.J.,Park, T.G.,Ko, K.S.,Kim, S.W. Elsevier Science Publishers 2010 Journal of controlled release Vol.143 No.1

A membrane receptor, Fas (CD95), and its ligand FasL have been considered as key players in diabetes pathogenesis. They are known to mediate interactions between β cells and cytotoxic T cells, which results in apoptotic cell death. We hypothesized that the interruption of Fas-FasL interactions by suppressing Fas expression in β cells would affect the development of diabetes. The effect of Fas-silencing siRNA (Fas siRNA) on diabetes development was evaluated in a cyclophosphamide (CY)-accelerated diabetes animal model after intravenous administration using a polymeric carrier, polyethylenimine (PEI). The systemic non-viral delivery of Fas siRNA showed significant delay in diabetes incidence up to 40days, while the control mice treated with naked Fas siRNA, scrambled dsRNA, or PBS were afflicted with diabetes within 20days. The retardation of diabetes incidence after the treatment of Fas siRNA may be due to the delayed progression of the pancreatic insulitis. In this study, the potential use of a non-viral carrier based siRNA gene therapy for the prevention of type-1 diabetes is demonstrated.

유전자 전달에서 양친매성 디펩티드들을 결합시킨 폴리아미도아민 덴드리머의 증대된 형질주입 효율

이제일(Jeil Lee),이선화(Sun Hwa Lee),최준식(Joon Sig Choi) 한국고분자학회 2018 폴리머 Vol.42 No.5

폴리아미도아민(PAMAM) 덴드리머는 널리 이용되는 비바이러스성 벡터 중 하나이며 생분해성 잔기를 가지고 있기 때문에 비교적 낮은 세포독성을 가지고 있는 것으로 알려져 있다. 그러나 빈약한 유전자 전달 능력은 폴리아미도아민 덴드리머의 생체 내 연구 및 임상적용에 장벽으로 작용하고 있다. 이러한 문제를 극복하기 위해서 폴리아미도아민 덴드리머 3차와 4차에 양친매성의 디펩티드인 라이신과 페닐알라닌을 도입하고 형질주입 능력을 평가하였다. 표면개질되지 않은 폴리아미도아민 덴드리머 3차, 4차와 비교할 때, 라이신과 페닐알라닌을 도입한 폴리아미도아민 덴드리머 4차는 HeLa, HepG2 세포 주에서 향상된 형질주입 효율을 나타내었고 또한 비교적 낮은 세포독성을 나타내었다. 이러한 결과들은 고분자들에 라이신, 페닐알라닌 디펩티드의 도입은 빈약한 유전자 전달 능력을 향상시킬 수 있음을 암시한다. Polyamidoamine (PAMAM) dendrimer consisting of biodegradable functional groups is one of the non-viral vectors and has showed relatively low cytotoxicity. However, its poor transfection efficiency acts as a barrier of in vivo research and medical application. To overcome this problem, we introduced amphiphilic KF dipeptides to PAMAM generation 3 and 4, and evaluated transfection efficiency. Compared with those of native PAMAM generation 3 and 4, PAMAM generation 4-KF showed enhanced transfection efficiency in HeLa and HepG2 cell lines. These results imply that introduction of KF dipeptide to polymers can improve the transfection efficiency.

Gene Therapy for Bone Tissue Engineering

김영동,Prasad Pofali,박태은,비제이,조기현,Sushila Maharjan,Prajakta Dandekar,Ratnesh Jain,최윤재,Rohidas B. Arote,조종수 한국조직공학과 재생의학회 2016 조직공학과 재생의학 Vol.13 No.2

Gene therapy holds a great promise and has been extensively investigated to improve bone formation and regeneration therapies in bone tissue engineering. A variety of osteogenic genes can be delivered by combining different vectors (viral or non-viral), scaffolds and delivery methodologies. Ex vivo & in vivo gene enhanced tissue engineering approaches have led to successful osteogenic differentiation and bone formation. In this article, we review recent advances of gene therapy-based bone tissue engineering discussing strengths and weaknesses of various strategies as well as general overview of gene therapy.

Human mesenchymal stem cells modification via photosensitizer to secret ligand inducing tumor targeted cell death

박소연,한지은,황희숙,나건 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-

In gene therapy, the delivery of gene to the target site is the most important. However, the gene therapy has a disadvantage, off-target effect, of causing inordinate biodistribution to major organs. The off-target effect can cause side effect such as normal cell death and decrease therapeutic effect. To complement for drawback, human mesenchymal stem cells (hMSCs) were used because hMSCs can migrate to tumor site by homing effect. Also, we used polymer as non-viral vector and the polymer was complexed with gene (g/plex). The gene expresses tumor targeted cell death ligands that kills tumor cells through apoptosis. However, the transfection efficiency of the g/plex was lower than viral vector. Therefore, we utilized a photosensitizer to enhanced efficiency of the g/plex transfection into hMSCs. Overall, cell death ligand expressing gene was successfully transfected into hMSCs via complexation and photosensitizer, so that hMSCs were modified to secrete cell death ligands at tumor site.

Charge reduction: an efficient strategy to reduce toxicity and increase the transfection efficiency of high molecular weight polyethylenimine

Mohammad Ramezani,Reza Kazemi Oskuee,Mehdi Rezaee,Leila Gholami,Mojtaba Seddighi Gildeh 한국약제학회 2019 Journal of Pharmaceutical Investigation Vol.49 No.1

With high potency in treatment of various diseases, gene therapy is mainly hindered by lack of safe and efficient gene delivery vectors. The current study was aimed to develop an efficient non-viral vector with adequate cytotoxicity. To this end, alkylcarboxylate chains (6C, 10C, 16C) were exploited to ameliorate the characteristics of PEI 750 kDa. Briefly, alkylcarboxylate chains with three different lengths (6C, 10C, 16C) were chemically grafted to the primary amine groups of PEI 750 kDa in three percentages (10, 50, and 100%). After evaluating the physicochemical properties of prepared vectors including surface charge, size, buffering capacity, and DNA condensing, their transfection efficiency and cytotoxicity were investigated in Neuro2A cells. The polyplexes size were 158.9–264.5 nm and their zeta potentials were 14–30 mV, while their buffering capacity and DNA condensing were not significantly decreased. The highest transfection efficiency in term of C/P ratio was observed in PEI750-10C-68%, PEI750-10C-7%, and PEI750-6C-7% at C/P ratios of 2, 4, and 6, respectively. Altogether, the decanoylcarboxylate-modified PEI with medium grafting percentages showed promising results as gene delivery vector. To sum up, the modification of high molecular weight PEIs by alkylcarboxylate chains is an efficient approach for development of more efficient non-viral vectors.