발표논문

• - Kim BK, Lee SJ, Kim TW, Kim H, Lee H, Hong EY, Cho YJ, Kwon S*(*Equal Corresponding authorship), Kim SH*. “EGT022, an RGD-containing recombinant disintegrin, inhibits the VEGF-induced angiogenic process by targeting integrin β3 in endothelial cells.” Am J Transl Res 2023; 15(3): 1831-41.
• - Hong, H. C., Kim, K. S., Park, S. A., Chun, M. J., Hong, E. Y., Chung, S. W., ... & Kim, S. H. “An mRNA vaccine against SARS-CoV-2: Lyophilized, liposome-based vaccine candidate EG-COVID induces high levels of virus neutralizing antibodies.” bioRxiv 2021; 2021-03.
• - Park, S.-Y., J. Y. Song, M. J. Lee, K. M. Lee, M. H. Jeon, R. J. Ko, S.-H. Kim and B. J. Kang. “Diagnostic Performance of Shear Wave Elastography as Add-on Test in Thyroid Nodules: a Systematic Review and Meta-Analysis.” International Journal of Thyroidology 2018; 11(1): 31.
• - Chung, S., J.-S. Park and S.-H. Kim. “Saphenous vein occlusion with cyanoacrylate: a systematic review.” J. the Korean Med Assoc 2017; 60(6): 499.
• - Kim, S., Y. Lee and S.-H. Kim. “Safety and effectiveness of fecal microbiota transplantation: a systematic review.” J the Korean Med Assoc 2017; 60(9): 761.
• - Park, S. Y., S. Y. Kim, W. J. Choi, S.-H. Kim and S. G. Hong. “Diagnostic Accuracy and Detection Rate of Real-Time PCR for Detection of Group B Streptococcal Colonization in Pregnant Women: Systemic Review of Literature and Meta-Analysis.” Ann Clin Microbiol 2017; 20(2): 42.

연구분야

• mRNA vaccine, 대상포진 백신, 면역보조제, 안질환 치료제

Lecture Summary

• Development of mRNA Vaccine Platform: Based on the Experience of COVID-19 mRNA Vaccines
  Bong-Kyu Kim, Sunjong Kwon, Yang Je Cho, Seok-Hyun Kim
  
  R&D Center, EyeGene Inc., Goyang-si, Gyeonggi-do 10551, Republic of Korea
  
  Abstract
  
  코로나19 팬데믹 이전에는 약독화 생백신, 불활성화 사백신과 같은 1세대 백신과 아단위 백신, 펩타이드 백신, 바이러스 유사입자 백신과 같은 2세대 백신 플랫폼을 이용한 백신 개발이 주를 이루었으나 코로나19를 계기로 3세대 백신 플랫폼, 특히, mRNA 백신이 처음으로 상용화되어 차세대 백신 플랫폼으로 자리를 잡았다. 특히, 코로나19 팬데믹에 가장 효과적으로 대응하기 위하여 개발 및 사용된 mRNA 백신은 유행성 바이러스 질환에 대한 예방효과가 입증되어 향후 도래할 수 있는 새로운 바이러스 전염병에 가장 효과적으로 빠른 시간에 대응할 수 있는 플랫폼으로 여겨지고 있다. 아이진 주식회사에서는 코로나19에 대응하기 위하여 mRNA 백신 (EG-COVID, EG-COVARo)을 개발하였으며, 현재 국내에서 기초접종 임상 1상을 완료 후 mRNA 백신의 안전성을 확인하고 호주에서 추가접종으로 임상 1/2a상을 진행하고 있다. 아이진 주식회사의 mRNA 백신은 자체 개발한 양이온성 리포좀 (EG-R)을 전달체로 사용하여 시장에서 사용되고 있는 LNP 기반의 타 mRNA 백신들에 비해 안전성이 우수하며 또한 동결건조의 형태로 제조하여 냉장 조건에서 장기간 안정하여 보관 및 운송이 용이하다는 장점이 있다. 아이진 주식회사는 코로나19 mRNA 백신의 개발 경험을 바탕으로 계절성 독감백신 및 결핵백신을 개발 중에 있다. 현재 계절성 독감백신은 시험 동물을 이용한 감염억제시험에서 그 효능을 확인하였으며, 결핵백신은 효능을 확인하기 위하여 조만간 시험 동물을 이용한 확인 시험을 개시할 예정이다. 이를 통해서 아이진의 mRNA 백신이 여러 질환에 대응할 수 있는 플랫폼으로서의 능력을 확인하고 향후 새로운 백신들을 mRNA 백신으로 개발하는데 적용하고자 한다.
  

발표논문

• 1. J. H. Park#, S. W. Hong#, S. Yun, D.-k. Lee*, and C. Shin*, Effect of an siRNA with asymmetric RNA/dTdT overhang on RNA interference activity. Nucleic Acid Ther 24, 364-371 (2014)
• 2. S. W. Hong#, J. H. Park#, S. Yun, C. H. Lee, C. Shin*, and D.-k. Lee*, Effect of the guide strand 3'-end structure on the gene silencing potency of asymmetric siRNA. Biochem J 461, 427-434 (2014)
• 3. S. W. Hong, Y. Jiang, S. Kim, C. J. Li, and D.-k. Lee*, Target gene abundance contributes to the efficiency of siRNA-mediated gene silencing. Nucleic Acid Ther 24, 192-198 (2014)
• 4. T. Y. Lee#, C. I. Chang#, D. Lee, S. W. Hong, C. Shin, C. Li, D. Haussecker, S. Kim, and D.-k. Lee*, RNA interference-mediated simultaneous silencing of four genes using cross-shaped RNA. Mol Cells 35, 320-326 (2013)
• 5. C. Chang, S. W. Hong, P. Dua, S. Kim, and D.-k. Lee*, The design, preparation, and evaluation of asymmetric small interfering RNA for specific gene silencing in mammalian cells. Method Mol Biol 942, 135-152 (2013)
• 6. C. I. Chang, T. Y. Lee, S. Kim, X. Sun, S. W. Hong, J. W. Yoo, P. Dua, H. S. Kang, S. Kim, D. Lee, C. Shin, C. J. Li, and D.-k. Lee*, Enhanced intracellular delivery and multi-target gene silencing triggered by tripodal RNA structures. J Gene Med 14, 138-146 (2012)
• 7. S.-g. Jo#, S. W. Hong#, J. W. Yoo, C. H. Lee, S. Kim, S. Kim, and D.-k. Lee*, Selection and optimization of asymmetric siRNA targeting human c-MET gene. Mol Cells 32, 543-548 (2011)
• 8. J. W. Yoo, S. W. Hong, S. Bose, H. Kim, S. Y. Kim, S. Kim, and D.-k. Lee*, The role of transglutaminase in double-stranded DNA-triggered antiviral innate immune response. Bull Kor Chem Soc 32, 3893-3898 (2011)
• 9. S. W. Hong, S. M. Hong, J. W. Yoo, Y. C. Lee, S. Kim, J. T. Lis, and D.-k. Lee*, Phosphorylation of the RNA polymerase II C-terminal domain by TFIIH kinase is not essential for transcription of Saccharomyces cerevisiae genome.Proc Natl Acad Sci USA 106, 14276-14280 (2009) [selected by Faculty of 1000]
• 10. C. I. Chang, J. W. Yoo, S. W. Hong, S. E. Lee, H. S. Kang, X. Sun, H. A. Rogoff, C. Ban, S. Kim, C. J. Li, and D.-k. Lee*, Asymmetric shorter-duplex siRNA structures trigger efficient gene silencing with reduced non-specific effects. Mol Ther 17, 725-732 (2009)
• 11. S. W. Hong#, M. Jung#, E. Kim, D.-k. Lee*, and S. Kim*, Identification of B52-dependent gene expression signature and alternative splicing using a D. melanogaster B52-null mutant. Bull Kor Chem Soc 4, 323-326 (2009)
• 12. S. W. Hong#, J. W. Yoo#, H. S. Kang, S. Kim, and D.-k Lee*, HIF-1alpha dependent gene expression program during the nucleic acid-triggered antiviral responses. Mol Cells 27, 243-250 (2009)
• 13. S. W. Hong, S. Kim, and D.-k. Lee*, The role of Bach2 in nucleic acid-triggered antiviral innate immune responses. Biochem Biophys Res Commun 365, 426-432 (2008)
• 14. C. I. Chang, S. W. Hong, S. Kim, and D.-k. Lee*, A structure-activity relationship study of siRNAs with structural variations. Biochem Biophys Res Commun 359, 997-1003 (2007)
• 15. J. W. Yoo#, S. W. Hong#, S. Kim, and D.-k. Lee*, Inflammatory cytokine induction by siRNAs is cell type- and transfection reagent-specific. Biochem Biophys Res Commun 347, 1053-1058 (2006)

연구분야

• mRNA therapeutics/vaccine, siRNA therapeutics

Lecture Summary

• Development of Advanced mRNA Chemistry Optimized for Therapeutic Applications
  
  Kyeongmi Kim¹, Younggyu Kang¹, Jinyoung Han¹, Jaejin Lee¹, Kyeongmin Lee¹ and Sun Woo Hong¹,*
  ¹mCureX Therapuetics, Inc., Korea
  
  Abstract
  
  Upon the first approval and commercialization of mRNA vaccine under the COVID-19 pandemic, mRNA medicine showed prominent expansion of target indication area including vaccines and therapeutics. Modified nucleosides, one of the key components of mRNA therapeutics technology, are thought to be essential to avoid innate immune responses that are triggered by foreign mRNAs. Here, we demonstrate a proprietary mRNA platform technology harnessing alternate modified nucleoside (Mod A) instead of conventional chemistry (N1-methylpseudouridine, m1Ψ). Series of the studies showed that Mod A-modified mRNAs barely induce the innate immune response, while achieving high protein expression. Reduced induction of cytokines in Mod A-modified mRNA treated mice in comparison to m1Ψ-modified mRNA was observed and, consistently, systemic toxicities determined by clinical signs, completed blood count and liver enzyme blood chemistry were significantly lower with Mod A-modified mRNAs compared to m1Ψ-modified mRNAs. COVID-19 vaccine candidate consists of Mod A-modified mRNA had potent in vivo activity demonstrated by the induction of neutralizing antibodies against SARS-CoV-2 viruses and a strong protection efficacy in COVID-19 mouse model. These data provide an idea that Mod A-modified mRNAs have distinct advantages over the conventional mRNAs in the perspective of safety, the critical element for the development of therapeutics which usually require higher dose level and repeated administrations.
  

발표논문

• 뎅기 바이러스 특이적 siRNA, 그러한 siRNA를 포함하는 이중나선 올리고 RNA 구조체 및 이를 포함하는 뎅기 바이러스 증식 억제용 조성물 (2014.07.04. 출원/ 대한민국, 미국, 호주 등록)
• 핵산 전달용 지질 나노입자 및 조성물 (2021.03.11. 출원)
• 생분해성 결합을 포함하는 이온화 지질 및 이를 포함하는 지질나노입자 (2022.01.17. 출원; 2023.01.16 PCT 출원)
• 폐를 표적장기로 하는 생분해성 지질나노입자 약물 전달 제형 및 이의 활용 (2022.10.27 출원)
• 신규 핵산 분자 (2022.02.07. 국내우선권주장 출원/등록; 2023.08.07 미국, 일본 국제출원; 2023.09 유럽 출원; 2023. 10 중국 예정)
• 신규 핵산 분자 제조용 플랫폼 (2022. 08. 10 출원)
• 3D 프린터 기반 일체형 미세유체 혼합기 (2022. 09. 30 출원)

연구분야

• mRNA LNP 플랫폼 연구 및 개발
• mRNA 공정연구 및 개발
• 자체 신약 COVID-19 mRNA vaccine STP2104, 2250 개발 및 임상 개발

Lecture Summary

• ST Pharm's Quick Response Strategy for Next-Pandemic with Leveraging Novel SmartCap® and STLNP® Platform Technology
  
  Joo-Sung Yang
  
  Biotechnology R&D, ST Pharm, Seoul, Korea
  
  Abstract
  
  Since 2018, ST Pharm has developed its own mRNA platform technology. Novel 5’-cap analogs, marketed as SmartCap^®, and lipid-nanoparticle (LNP) drug delivery system, known as STLNP® have been introduced with strong IP position.
  Series of 5’-capping derivatives (ca. >30) with different ribose and base combination are available for capping screening purpose. The in-house capping library screening system is providing our clients for identifying the best capping analogue with high capping efficiency and potency. Also, the optimized capping analogue will be specifically applicable to the desired target cell/organ. The novel ionizable lipids used for STLNP^® formulation showed similar physicochemical property and performance to other well-known LNPs. Some of the STLNP^® is delivered to a specific organ, as determined by in vivo biodistribution. Mass production of key raw materials for SmartCap^® and STLNP^® is feasible utilizing CFT and SMB technology. ST Pharm can also manufacture both non-GMP and GMP-grade 5’-capping reagents (> kgs/year) and the ionizable and PEG-lipid in LNP (> tons/year). Our seamless mRNA platform technologies are highly cost competitive and enabling customers for their successful mRNA therapeutics and vaccine development.
  Herein, we are presenting in-house experience and know-hows of SmartCap^® and STLNP^®, and their applications to mRNA COVID-19 vaccine programs that are currently in clinical trials, and Expedite 100-day project for rapid preparedness for new/emerging infectious diseases.
  
  

발표논문

• • Jonathan F. Lovell, Yeong Ok Baik, Seuk Keun Choi, Chankyu Lee, Jeong-Yoon Lee, Kazutoyo Miura, Wei-Chiao Huang, Young-Shin Park, Sun-Je Woo, Sang Hwan Seo, Jae-Ouk Kim, Manki Song, Chung-Jong Kim, Jae-Ki Choi, Jieun Kim, Eun Ju Choo & Jung-Hyun Choi. Interim analysis from a phase 2 randomized trial of EuCorVac-19: a recombinant protein SARS-CoV-2 RBD nanoliposome vaccine. BMC Medicine. 2022.
• • Young Jae Lee, Ji Eun Yu, Paul Kim, Jeong-Yoon Lee, Yu Cheol Cheong, Yoon Jae Lee, Jun Chang, Baik Lin Seong. Eliciting unnatural immune responses by activating cryptic epitopes in viral antigens. FASEB Journal. 2018.
• • Yo Han Jang, Joo Young Kim, Young Ho Byun, Ahyun Son, Jeong-Yoon Lee, Yoon Jae Lee, Jun Chang, Baik Lin Seong. Pan-influenza A protection by prime-boost vaccination with cold-adapted live attenuated influenza vaccine in a mouse model. Frontiers in immunology. 2018.
• • Young J. Lee†, Jeong Y. Lee†, Yo H. Jang, Sang-Uk Seo, Jun Chang, Baik L. Seong. Non-specific effect of vaccines: Immediate protection against respiratory syncytial virus infection by a live attenuated influenza vaccine. Frontiers in microbiology. 2018. (co-first author).
• • Lee JY, Chang J. Recombinant baculovirus-based vaccine expressing M2 protein induces protective CD8+ T-cell immunity against respiratory syncytial virus infection. J Microbiol. 2017.
• • Lee JY, Chang J. Universal vaccine against respiratory syncytial virus A and B subtypes. PLoS One. 2017.
• • Lee HJ, Lee JY, Park MH, Kim JY, Chang J. Monoclonal Antibody against G Glycoprotein Increases Respiratory Syncytial Virus Clearance In Vivo and Prevents Vaccine-Enhanced Diseases. PLoS One. 2017.

연구분야

• 바이러스 면역, 백신

Lecture Summary

• A Novel Nanoliposome Adjuvant Platform for Infectious Diseases Vaccine Development
  Jeong-Yoon Lee
  
  Eubiologics Co., Ltd., Chuncheon, South Korea
  
  Abstract
  
  Concerns about the emergence of a pandemic are growing as the incidence and cycle of new viral infectious diseases becomes shorter over the world. While dealing with new infectious diseases such as SARS, H1N1 flu, MERS, and, most recently, COVID-19, it is critical to secure vaccine technologies and platforms that can quickly respond to new emerging infectious diseases. Eubiologics has been developing a novel nanoparticle adjuvant platform for use in the development of recombinant protein vaccines against various viral diseases. This nanoparticle adjuvant is based on liposome with EcML (monophosphoryl lipid A (MPLA) derived from genetically engineered E. coli, TLR-4 agonist) and SNAP (Spontaneous Nanoliposome-Antigen Particleization) technology, which can display antigens on the liposome surface. EuCorVac-19, a COVID-19 vaccine developed by Eubiologics, is made of the SARS-CoV-2 receptor-binding domain (RBD) and a nanoparticle adjuvant platform, and its remarkable efficacy and safety have been proven in phase 1/2 clinical trials. In addition, phase 3 clinical trial is in progress, with intermediate results demonstrating superiority (immunogenicity) and non-inferiority (sero response rate) compared to the comparator vaccine. This adjuvant platform has the advantage of being adaptable to the development of viral vaccines by appropriately replacing the main vaccine antigens of different viruses. As a result, in addition to the COVID-19 vaccine, it is being applied to develop other virus vaccines such as RSV and VZV. Moreover, it is expected to contribute to the development of other novel vaccines in combination with MPLA.
  

발표논문

• Choi HK, Kim SI, Son JS, Hong Ss, Lee HS, Lee HJ. Enhancement of paclitaxel production by temperature shift in suspension culture of Taxus chinensis. Enzyme Microb. Technol. 2000; 27: 593 -598
• Choi HK, Kim SI, Son JS, Hong SS, Lee HS, Chung IS, Lec HJ. Intermittent maltose feeding enhances paclitaxel production in suspension culture of Taxus chinensis cells. Biotechnol Lett 2000; 22: 1793 – 1796
• Kim SI, Choi HK, Son JS, Yun JH, Jang MS, Kim HR, Song JY, Kim JH, Choi HJ, Hong SS. Cryoprescrvation of Taxus chinensis suspension cell cultures. CryoLetters 2001; 22: 43 – 50
• Son JS, Choi HK, Hong SS. Recovery of extracellular paclitaxel from suspension culture of Taxus chinensis by adding inorganic salts. Biotechnol. Lett. 2001; 23: 723 – 725
• Choi HK, Kim SI, Son JS, Kim HR, Song JY, Kim JH, Hong SS. Enhanced production of paclitaxel by semi-continuous batch process (SCBP) in suspension culture of Taxus chinensis. Enzyme Microb. Technol. 2001; 29: 583 – 586
• Choi HK, Son JS, Na KH, Hong SS, Song JY. Mass production of paclitaxel with plant cell culture. Korea Journal of Plant Biotechnology. 2002; 29: 59-62
• 원료의약품 제조 및 품질관리기준 (GMP) 가이던스 – 식품의약품 안전처

연구분야

• 원료의약품의 상업화를 위한 식물세포배양 공정의 개발과 스케일업을 수행하였다. 플라스크 수준의 개발, 3000리터 파이롯트 스케일업을 거쳐 32000리터 상업 생산 스케일의 가동에 이르는 전체 상업화 과정의 공정 개발, 문제점 분석 등을 실시하였다. 연구개발에 참여한 제품에 대해 국내 및 해외 의약품 등록 업무를 수행하고 품질관리 업무를 수행하여 의약품 개발 – 허가 – 생산 - 판매에 이르는 전 life-cycle에 관여하고 있다.

Lecture Summary

• Construction of Facilities and Scale-up of mRNA Production
  
  Joo-Sun Son* and Dong-Myeong Lee
  
  R&D Center, BMI Korea Co., Ltd.,
  
  Abstract
  
  코로나19 백신 및 mRNA 원부자재 생산시설·장비 확충사업으로 ㈜한국비엠아이 오송공장에 GMP 시설을 구축 및 문서화를 진행하였고, mRNA와 mRNA 전달체인 양이온성 리포좀 생산을 위한 장비(생산 및 품질)를 설치 후 적격성평가를 실시하였고, 이후 시험생산을 통해서 GMP 시설, 장비(생산, 품질) 및 품질에 이상이 없음을 확인하였다. 현재 ㈜한국비엠아이 오송공장에서는 최대 30L까지 mRNA 생산이 가능하다. 그리고 백신용 mRNA 원료의약품 GMP급 대량생산 과제를 통해서 mRNA 원료의약품 생산공정에 대해서 원부자재(대체 원부자재 포함) 및 공정 최적화를 통해서 IPC 항목 설정, 공정 및 품질 시험방법을 검증하고 있으며, 최적화 후 제조단위를 스케일-업을 진행하면서 백신용 mRNA 원료의약품 GMP급 대량생산을 하기 위해서 mRNA 플랫폼을 구축하여 향후 백신으로 이용가능한 수천 dose 분량의 mRNA 생산을 위한 공정을 개발 중이다.
  

발표논문

• Safety and Immunogenicity of the ID93 + GLA SE Tuberculosis Vaccine in BCG Vaccinated Healthy Adults: A Randomized, Double Blind, Placebo Controlled Phase 2 Trial

연구분야

• 백신 및 면역치료제

Lecture Summary

• Status of next generation vaccine development by Quratis
  
  Kwan Goo Cho^{1
  1}Ceo and Chairman of Quratis Inc, Seoul, Korea
  
  Abstract
  
  -General company introduction and history
  - Next genetation TB vaccine development
  - Next genetation mRNA Covid-19 vaccine
  - New parasite vaccine for Schistosomiasis
  

발표논문

• Mattoo, S.S., Kim, S.J., Ahn, D.G., Myoung, J. 2022. Escape and over-activation of innate immune responses by SARS-CoV-2: two faces of a coin. Viruses (IF: 5.048). 14(3): 530
• Mattoo, S.S., Myoung, J. 2021. A promising vaccination strategy against COVID-19 on the horizon: heterologous immunization. Journal of Microbiology and Biotechnology (IF: 3.45). 31(12): 1601-1614
• Lee, S., Yoon, G.Y., Myoung, J. Kim, S.J., Ahn, D.G. 2020. Robust and persistent SARS-CoV-2 infection in the human intestinal brush border expressing cells. Emerging Microbes and Infections (IF: 5.828). 9(1):2169-2179
• Lee, J., Bae, S., Myoung, J. Generation of full-length infectious cDNA clones of Middle East respiratory syndrome coronavirus. Journal of Microbiology and Biotechnology (IF: 1.992). 29(6):999-1007.
• Park, M.K., Cho, H.J., Rho, S.W., Kim, S.J., Myoung, J. 2019. Cell-type-specific IFN-gamm-mediated inhibition of Kaposi’s Sarcoma-associated Herpesvirus. Scientific Reports (IF: 4.7). 10(1): 1-12.

연구분야

• Coronavirus (SARS-CoV-2, MERS-CoV): host-pathogen interactions and vaccines

발표논문

• Mattoo, S.S., Kim, S.J., Ahn, D.G., Myoung, J. 2022. Escape and over-activation of innate immune responses by SARS-CoV-2: two faces of a coin. Viruses (IF: 5.048). 14(3): 530
• Mattoo, S.S., Myoung, J. 2021. A promising vaccination strategy against COVID-19 on the horizon: heterologous immunization. Journal of Microbiology and Biotechnology (IF: 3.45). 31(12): 1601-1614
• Lee, S., Yoon, G.Y., Myoung, J. Kim, S.J., Ahn, D.G. 2020. Robust and persistent SARS-CoV-2 infection in the human intestinal brush border expressing cells. Emerging Microbes and Infections (IF: 5.828). 9(1):2169-2179
• Lee, J., Bae, S., Myoung, J. Generation of full-length infectious cDNA clones of Middle East respiratory syndrome coronavirus. Journal of Microbiology and Biotechnology (IF: 1.992). 29(6):999-1007.
• Park, M.K., Cho, H.J., Rho, S.W., Kim, S.J., Myoung, J. 2019. Cell-type-specific IFN-gamm-mediated inhibition of Kaposi’s Sarcoma-associated Herpesvirus. Scientific Reports (IF: 4.7). 10(1): 1-12.

연구분야

• Coronavirus (SARS-CoV-2, MERS-CoV): host-pathogen interactions and vaccines

Lecture Summary

• Investigation of Tanscription Regulatory Sequence-mediated viral gene expression by utilizing reverse genetics system and its application to vaccine development
  
  
  Jinjong Myoung, Ph.D.
  Korea Zoonosis Research Institute, Jeonbuk National University
  
  Abstract
  
  The emergence of SARS-CoV-2, initially brought to the attention of the World Health Organization on December 31, 2019, presented an unparalleled challenge to global public health. Since its initial identification, novel variants have arisen, supplanting preceding variants and protracting the duration of the global health crisis. At the outset, the research primarily centered around the development of vaccines and treatments. Nevertheless, there is a gradual transition occurring towards an enhanced comprehension of the molecular mechanisms that drive the pathogenesis of SARS-CoV-2. The manipulation of SARS-CoV-2 genetics necessitates the utilization of reverse genetics, a technique that involves the conversion of RNA into cDNA. This process is essential due to the virus's single-stranded RNA nature and its possession of the largest genome among RNA viruses, measuring 32 kb. Reverse genetics enables the application of diverse molecular biological techniques in the study of SARS-CoV-2. In this study, full-length complementary DNA (cDNA) copies of SARS-CoV-2 were generated utilizing a range of promoters. Furthermore, we incorporated reporter genes (mNeonGreen or mCherry) into the viral genome using various arrangements in order to ascertain the most effective genetic configurations that facilitate proficient expression of the inserted reporter genes. This measure was implemented to enhance the monitoring of viral infections in both laboratory-controlled and live organism environments. Our study aimed to investigate the mechanisms involved in the regulation of viral transcription through transcription regulatory sequences (TRS), using SARS-CoV-2 cDNA constructs. Our study has effectively shown that both the transcription start site (TRS) and the 5' flanking sequences play crucial roles in facilitating the optimal expression of downstream genes. Conversely, we have found that the 3' flanking sequences, if present, have minimal or insignificant effects on gene expression. The mutant viruses, in which a reporter gene was inserted between TRS and 5' flanking sequences, exhibited a diminished phenotype in both Vero E6 cells and Calu-3 cells when compared to the wild-type virus. It is noteworthy that there was a notable impairment in the overall expression of viral genes, resulting in a significant reduction in viral protein levels. At present, the application of TRS-based attenuation is being utilized in the rational design of coronavirus vaccines.
  

발표논문

• 1. Hyun Jung Hwang and Yoon Ki Kim (2023.02) The role of LC3B in autophagy as an RNA-binding protein. Autophagy 19(3): 1028-1030.
• 2. Ki-Jun Yoon and Yoon Ki Kim (2022.11) Resolving m6A epitranscriptome with stoichiometry. Trends in Genetics 38(11):1099-1100.
• 3. Sung Ho Boo, Hongseok Ha, and Yoon Ki Kim (2022.09) m1A and m6A modifications function cooperatively to facilitate rapid mRNA degradation. Cell Reports 40(10):111317.
• 4. Sung Ho Boo, Hongseok Ha, Yujin Lee, Min-Kyung Shin, Sena Lee, and Yoon Ki Kim (2022.05) UPF1 promotes rapid degradation of m6A-containing RNAs. Cell Reports 39(8):11086.
• 5. Hyun Jung Hwang, Hongseok Ha, Ban Seok Lee, Bong Heon Kim, Hyun Kyu Song, and Yoon Ki Kim. (2022.03) LC3B is an RNA-binding protein to trigger rapid mRNA degradation during autophagy. Nature Communications 13:1436.

연구분야

• mRNA decay, mRNA modification, circular RNA, translation mechanism

Lecture Summary

• Molecular features of lariat-capped RNA and its application
  Yoon Ki Kim¹, *
  ¹Department Department of Biological Sciences, KAIST
  
  Abstract
  In an effort to combat the global pandemic caused by the coronavirus, many pharmaceutical companies are striving to develop coronavirus vaccines. Notably, a groundbreaking new type of vaccine that utilizes mRNA has recently been developed and is being commercialized for the first time in history. mRNA-based vaccines have a significant advantage due to their ease of manufacture, enabling rapid responses to various infectious diseases. However, due to the inherent instability of mRNA itself, there are technological limitations in its utilization. In our lab, we are currently developing lariat-capped RNA, a novel type of RNA with a high stability. Unlike the general 5'-cap structure, lariat-capped RNA is highly stable within cells. The use of ribozymes makes it easy to produce lariat-capped RNA through in vitro transcription. More importantly, while there are numerous patents for various known techniques to increase mRNA stability, there are currently no patents related to lariat-capped RNA. Therefore, given the advantages mentioned earlier, we anticipate that lariat-capped RNA technology can be used to develop a versatile mRNA vaccine platform for not only the coronavirus but also various infectious diseases.
  

발표논문

• Zou Y et al. Commun Biol, 2023, 6(1):272.
• Lee AR et al. J Microbiol Biotechnol, 2022, 32(8):955-959.
• Choi H et al. Front Microbiol, 2022, 13:891646.
• Kim C et al. J Microbiol, 2022, 60(3):335-346.
• Lim J et al. Nanomedicine, 2021, 37:102438.

연구분야

• Respiratory infection
• Innate immunity
• Vaccine
• Japanese encephalitis
• Commensal microorganism

Lecture Summary

• Leveraging Escherichia coli-based antigen expression for rapid response to emerging infectious diseases Sang-Uk Seo
  
  Department of Microbiology, College of Medicine, The Catholic University of Korea
  
  Abstract
  
  Emerging infectious diseases continue to threaten global public health, necessitating innovative strategies for the rapid development of prophylactic measures. Escherichia coli, a well-established and easily manipulable microorganism, serves as a powerful biotechnological tool for cost-effective and scalable recombinant antigen production. While E. coli-based systems hold great promise, they come with certain challenges, notably the often insoluble expression of antigens. One prominent approach involves the conjugation of target antigens with RNA-interacting domains, resulting in a significant increase in their solubility within the E. coli-based system. Additionally, the utilization of scaffold structures capable of generating nanoparticles enhances the efficiency of vaccine-induced immune responses. Despite the inherent weaknesses of the E. coli system when compared to eukaryotic expression, advanced techniques render it increasingly practical and useful. Especially in emergent situations like recent pandemics, E. coli-based vaccine development holds merits as a valuable and rapid-response option.
  

발표논문

• 2023년 교신저자
• 1. Understanding nucleic acid sensing and its therapeutic applications. Kong LZ, Kim SM, Wang C, Lee SY, Oh SC, Lee S, Jo S, Kim TD. Experimental and Molecular Medicine. In Press [correspondence, IF 12.172]
• 2. NK cells encapsulated in micro/macropore-forming hydrogels via 3D bioprinting for tumor immunotherapy. Kim D, Jo S, Lee D, Kim SM, Seok JM, Yeo SJ, Lee JH, Lee JJ, Lee K, Kim TD*, Park SA*. Biomaterials Research. 2023 Jun 22; 27(1):60 (* equal correspondence). [correspondence, IF 15.863]
• 3. FTO negatively regulates NK cell killing efficacy. Kim SM, Oh SC, Lee SY, Kong LZ, Lee JH, Kim TD. EMBO Rep. 2023 Apr 5;24(4): e55681. [Correspondence, IF 9.07 ]
• 4. NgR1 is an NK cell inhibitory receptor that destabilizes immunological synapse. Oh SC, Kim SE, Jang IH, Kim SM, Lee SY, Lee SY, Chu IS, Yoon SR, Jung HY, Choi I, Doh JS*, Kim TD*. Nat. Immunol. 2023 Mar; 24(3):463 (* equal correspondence). [Correspondence, IF 31.25]
• 5. A modifiable universal cotinine-chimeric antigen system of NK cells with multiple targets. Kang HY, Lee SY, Kim HM, Lee H, Cho MY, Park HS, Han EH, Chung J, Hong KS*, Choi I*, Kim TD*. Frontiers in Immunol. 2023 Jan. 13; 13:1089369 (* equal correspondence). [Correspondence, IF 8.786 ]

연구분야

• RNA biology, NK cell biology, gene therapy (RNA and CAR-NK)

Lecture Summary

• Development of mRNA vaccine platforms using novel synthetic molecular design
  Seok-Min Kim¹, Dongwon Yoo², and Tae-Don Kim¹*
  ¹ Immunotherapy Research Center, KRIBB, Daejeon, Korea
  ² School of Chemical and Biological Engineering, Seoul National University, Seoul, Korea
  
  Abstract
  
  As part of our research to develop a highly efficient mRNA vaccine platform based on novel molecular synthesis technology, we would like to introduce a new platform construction technology for mRNA vaccine technology that has recently attracted attention. Among them, we would like to describe the technical features and differentiation of a recently patented linear mRNA structure. We observed that the mRNA construct exhibits highly efficient protein expression and significantly higher antibody production against Cov2 spike antigen compared to the control mRNA vaccine from Pfizer. In addition, we synthesized novel LNPs and showed high mRNA delivery efficiency compared to the control. Based on these various new synthesis and design technologies, the technology to amplify the delivery efficiency and expression efficiency of cells is expected to be useful not only for increasing the efficacy of vaccines, but also for cancer vaccines and mRNA-based therapies.
  

발표논문

• SARS-CoV-2 variants with NSP12 P323L/G671S mutations display enhanced virus replication in ferret upper airways and higher transmissibility. Kim SM et al. Cell Rep. 2023.

• Enhanced antibody responses in fully vaccinated individuals against pan-SARS-CoV-2 variants following Omicron breakthrough infection. Jeong HW, et al. Cell Rep Med. 2022.

• Age-dependent pathogenic characteristics of SARS-CoV-2 infection in ferrets. Kim YI, et al. Nat Commun. 2022.

• Infection and Rapid Transmission of SARS-CoV-2 in Ferrets. Kim et al Cell Host Microbe. 2020

• Cross-genotype protection of live-attenuated vaccine candidate for severe fever with thrombocytopenia syndrome virus in a ferret model. Yu KM, et al PNAS, U S A. 2019,

• Ferrets animal model of severe fever with thrombocytopenia syndrome phlebovirus for human lethal infection and pathogenesis. Park et al Nat Microbiol. 2019

연구분야

• Emerging and Reemerging viruses, Zoonotic diseases, Vaccines

Lecture Summary

• Zoonotic viral disease and their pathogenic potential: pathogenesis mechanisms and control strategy
  Young-Il Kim ¹, Se-Mi Kim ¹, Eun-Ha Kim ¹, Mark Anthony B.Casel ^1,2, Rare Rollonl ^1,2, Seung-Gyu Jangl ^1,2, Young Ki Choil ^1,2,
  ¹ Center for Study of Emerging and Re-emerging Viruses, Korea Virus Research Institute, Institute for Basic Science (IBS), Daejeon 34126, Republic of Korea
  ² College of Medicine and Medical Research Institute, Chungbuk National University, Cheongju, Republic of Korea
  
  Abstract
  
  In light of recent pandemics, including influenza and the Coronavirus pandemic, viral diseases have emerged as one of the most imperative areas of research. Notably, there has been remarkable and rapid progress in the fields of novel diagnostics, mRNA vaccine technology, and the development of novel antiviral drugs. Nonetheless, numerous zoonotic diseases continue to pose a significant threat to human life. Specifically, the looming risk of pandemics arising from various zoonotic diseases, such as avian influenza viruses, SFTSV, and Disease-X, underscores the critical importance of the one-health concept. Today, I would like to provide an overview of the current state of zoonotic infectious diseases, present the most recent research findings, and discuss the latest advancements in the field. Specifically, I would like to pay attention to elucidating the pathogenic mechanisms and detailing the status of vaccine development for SFTSV, a recent emergence in Asia, as well as the highly pathogenic avian H5Nx influenza virus, which has evolved into a global concern. I hope that this discourse will establish a foundation for fostering communication and fostering collaborative research efforts with experts from diverse scientific disciplines
  

발표논문

• Chung, Hae-Rynn, Eun-Ah Kim, and Jun Chang. "A “Prime and Deploy” Strategy for Universal Influenza Vaccine Targeting Nucleoprotein Induces Lung-Resident Memory CD8 T cells."Immune Network 21.4 (2021): 28-28.
• Chang, Jun. "MHC multimer: A Molecular Toolbox for Immunologists." Molecules and Cells 44.5 (2021): 328-334.
• Chang, Jun. "Adenovirus Vectors: Excellent Tools for Vaccine Development." Immune Network 21.1 (2021): 6-6.
• MH Kim, HJ Kim, J Chang (2019) Superior immune responses induced by intranasal immunization with recombinant adenovirus-based vaccine expressing full-length Spike protein of Middle East Respiratory Syndrome Coronavirus. PLoS ONE 14(7), e0220196.
• MH Kim, JO Kang, JY Kim, HE Jung, HK Lee, J Chang (2019) Single mucosal vaccination targeting nucleoprotein provides broad protection against two lineages of influenza B virus. Antiviral Research 163, 19-28

연구분야

• Vaccine for respiratory virus infection, T-cell immunology

Lecture Summary

• T-cell-based Universal Influenza Vaccine
  
  Myung Hee Kim, Haerynn Chung, Gyeongjoo Park, Eun-Ah Kim, and Jun Chang*
  Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul, Korea.
  
  Influenza vaccines that target the highly variable surface glycoproteins hemagglutinin and neuraminidase cause inconvenience of having vaccination every year. For this reason, development of universal vaccines targeting conserved viral components is needed. In this study, we generated recombinant adenovirus (rAd) vaccine encoding nucleoprotein (NP) of influenza A virus (A/PR/8/34) and B virus (B/Yamagata/16/1988), designated rAd/A-NP and rAd/B-NP, respectively, and investigated their immunogenicity and protective efficacy against influenza virus infections. We found that NP-specific CD8 T cells induced by the vaccine played a major role in protection following influenza virus infection. To assess the effects of the administration routes on protective immunity, we immunized mice via intranasal or intramuscular routes. Both groups showed strong NP-specific humoral and CD8 T-cell responses, but only intranasal immunization provided complete protection against influenza virus challenge. Intranasal but not intramuscular administration established resident memory CD8 T cells in the airway and lung parenchyma, which were required for efficient protection. These findings demonstrate that non-replicating adenovirus-based NP vaccines could be developed as a universal vaccine for influenza.
  

발표논문

• 1. Kwak K*, Sohn H*, George R, Torgbor C, Manzella-LapeiraJ, Brzostowski J and Pierce SK. (2023) B cell responses to membrane-presented antigens require the function of the mechanosensitive cation channel Piezo1. Sci Signal. Accepted.
• 2. Notario GR, Kwak K. (2022) Increased B cell understanding puts improved vaccine platforms just over the horizon. Immune Netw., 10.4110/in.2022.22.e47.
• 3. Ambegaonkar AA, Kwak K, Sohn H, Manzella-Lapeira J, Brzostowski J, Pierce SK. (2020) Expression of inhibitory receptors by B cells in chronic human infectious diseases restricts responses to membrane- associated antigens. Sci Adv., 10.1126/sciadv.aba6493

연구분야

• B cell immunology, Vaccinology

Lecture Summary

• Antigen Valency Dictates the Strength and Quality of B Cell Response
  
  Seungyong Baek ¹, Minhee Ham ¹, Youngjae Hong ¹, Kihyuck Kwak¹*
  
  ¹ Department of Microbiology and Immunology, Institute for Immunology and Immunological Diseases, Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, Seoul 03722, Korea.
  
  Abstract
  
  Current human papilloma virus-virus-like particle (HPV-VLP) vaccines exhibit unrivaled efficacy, inducing long-lasting neutralizing antibody responses that provide protection against HPV infections and HPV-related cancers even after a single dose. Due to the robust immunogenicity exhibited by VLPs, the VLP platform is frequently employed in the formulation of vaccines targeting many other pathogens, including SARS-CoV-2 and Influenza. The cellular and molecular mechanisms underlying the ability of VLP vaccines to evoke a robust immune response, however, have not been thoroughly investigated. To better comprehend this mechanism, we developed an antigen system that can modify the valency of gp120 antigen (1mer, 4mer, 8mer, 60mer) as well as a mouse system consisting of B cells that express a BCR recognizing gp120 antigen. We discovered that high valency (HV) antigen (60mer) substantially induced BCR micro-clusters, whereas low valency (LV) antigen (4mer) induced BCR nano-clusters, and that the calcium flow induced by HV antigen was greater and lasted longer. Moreover, LV antigen preferentially activates Akt signaling, while HV antigen preferentially activates NF-kB and NFAT signaling, both of which are essential for optimal B cell activation. Bulk mRNA sequencing revealed that HV antigen induced greater expression of genes related to germinal center response than LV antigen, and also induced high levels of IL-2RA gene expression related to B cell proliferation, suggesting that HV antigen generates a signaling pathway that amplifies B cell activation. We hypothesize that the elucidation of the new signaling molecular targets that cause robust B cell activation by HV Antigen and the development of a compound that can activate these targets would make a significant contribution to vaccine development.
  

발표논문

• (1)Dengue virus-polymersome hybrid nanovesicles for advanced drug screening using real-time single nanoparticle virus tracking. ACS Applied Material& Interfaces, 2020. (Corresponding author)
• (2)Host Cell Mimic Polymersomes for Rapid Detection of Highly Pathogenic Influenza Virus via a Viral Fusion and Cell Entry Mechanism. Advanced Functional Materials, 2018. (Corresponding author)
• (3)Reactive Oxygen Species-Regulating Polymersome as an Antiviral Agent against Influenza Virus. Small, 2017. 13(32). (Corresponding author)
• (4)Experimental infection of clade 1.1.2 (H5N1), clade 2.3.2.1c (H5N1) and clade 2.3.4.4 (H5N6) highly pathogenic avian influenza viruses in dogs. Transbound. Emerg. Dis. 2017. 64(6); 1669-1675. (Corresponding author)
• (5)Attenuation of the virulence of a recombinant influenza virus expressing the naturally truncated NS gene from an H3N8 equine influenza virus in mice. Vet. Res. 2016. 15;47(1):115 (Corresponding author); Selected from 120 peer-reviewed articles

연구분야

• 바이러스, 인수공통감염병, 스필오버, 진단, 백신

Lecture Summary

• Nanobiotechnology for diagnosis and vaccine against Disease X; unexpected emerging viral diseases
  
  Daesub Song, Ph.D., D.V.M
  College of Veterinary Medicine, Seoul National University, Korea
  
  Abstract
  
  Unlike in the past, emergences of viral infections have been and are occurring at very frequent intervals, causing enormous deaths and disability worldwide. Against a constant background of established infections, periodical emerging the epidemics of highly pathogenic influenza viruses (HPAI) greatly expand the global burden of infections. Accurate and rapid diagnosis of viral infections can result in effective and appropriate prevention and quarantine measures. This study compares and discusses about rapid diagnostics for the differential patho-typing between HPAI and LPAI using nanobiotechnology. The field of nanotechnology encompasses those technologies to fabricate materials, including sphere, cubic and nanoscale particles. Therefore, nanobiotechnology has the potential to offer not merely advances in diagnostics and vaccination to control infectious diseases but also in delivering various capabilities as below; Highly pathogenic avian influenza virus (HPAIV) infections have occurred continuously and crossed the species barrier to humans, leading to fatalities. A PCR-based molecular test is currently the most sensitive diagnostic tool for HPAIV; however, the results must be analyzed in centralized diagnosis systems by a trained individual. This requirement leads to delays in quarantine and isolation. To control the spread of HPAIV, rapid and accurate diagnostics suitable for field testing are needed, and the tests must facilitate a differential diagnosis between HPAIV and low pathogenic avian influenza virus (LPAIV), which undergo cleavage specifically by trypsin- or furin-like proteases, respectively. In this study, we have developed a differential avian influenza virus (AIV) rapid test kit and evaluated it in vitro and using clinical specimens from HPAIV H5N1-infected dogs. We demonstrated that this rapid test kit provides highly sensitive and specific detection of HPAIV and LPAIV and is thus a useful field diagnostic tool for H5N1 HPAIV outbreaks and for rapid quarantine control of the disease. In addition to diagnostics, development of better adjuvant accompanied with vaccine for enhancing immunogenicity has been greatly required for the control of influenza infection. Herein, we also address nano-complex of amphiphilic grafted poly (amino acid) and hydrophobic squalene (PA/S-NC) as a potent adjuvant that can act as a robust strategy for induce humoral (Th2) and cellular (Th1) immune responses as well as a delivery agent of antigens. CASq performed great biocompatibility, particle stability, and produced a high degree of antigen-specific antibodies and T cell immune responses in mice when CASq was co-administered with inactivated whole influenza virus antigen (CA04), in which CASq exhibited complete protection against lethal infection. Keyword: HPAIV, nanobiotechnology, diagnostics, adjuvant, vaccination
  

발표논문

• Rhee JH, Khim K, Puth S, Choi Y, Lee SE. Deimmunization of flagellin adjuvant for clinical application. Curr Opin Virol. 2023 Apr 19;60:101330. doi:10.1016/j.coviro.2023.101330. Online ahead of print
• Puth S, Verma V, Hong SH, Tan W, Lee SE, Rhee JH. An all-in-one adjuvanted therapeutic cancer vaccine targeting dendritic cell cytosol induces long-lived tumor suppression through NLRC4 inflammasome activation. Biomaterials. 2022 May 10;286:121542. doi: 10.1016/j.biomaterials.2022.121542.
• Puth S, Hong SH, Na HS, Lee HH, Lee YS, Kim SY, Tan W, Hwang HS, Sivasamy S, Jeong K, Kook JK, Ahn SJ, Kang IC, Ryu JH, Koh JT, Rhee JH, Lee SE. A built-in adjuvant-engineered mucosal vaccine against dysbiotic periodontal diseases. Mucosal Immunol. 2019 Mar;12(2):565-579. doi: 10.1038/s41385-018-0104-6. Epub 2018 Nov 28. (co-correspondence)

연구분야

• Molecular microbial pathogenesis/Vaccinology/Immunotherpy

Lecture Summary

• Flagellin: A Versatile Engineerable Vaccine Adjuvant
  
  
  Joon Haeng Rhee
  
  Department of Microbiology and Combinatorial Tumor Immunotherapy MRC and Clinical Vaccine R&D Center, Chonnam National University Medical School, Hwasun, Korea
  
  Modern vaccines became safer than old conventional vaccines by improving purity and regulating composition with defined constituents. Consequently, new vaccines confer lower immunogenicity because of the removal of built-in adjuvant components through more refined purification manufacture process. Hence, newly developed vaccines require coformulation with effective adjuvants. TLR ligands are considered attractive adjuvants for vaccines and immunotherapy. Flagellin is the cognate ligand for Toll-like receptor 5 (TLR5) of host cells. TLR stimulation leads to activation of innate immunity and subsequently modulates adaptive immune responses. Flagellin has an excellent adjuvanticity for mucosal vaccines. Mucosal co-administration of a V. vulnificus flagellin (FlaB) with microbial antigens induced significantly enhanced antigen-specific IgA responses in both mucosal and systemic compartments and IgG responses in the systemic compartment. Intranasally administered FlaB targets TLR5 expressing CD11c+ DCs in draining lymph nodes and stimulate induction of antigen-specific T and B cell responses. Further, FlaB served an efficacious adjuvant for cancer vaccines, which could be successfully combined with conventional therapies such as radiation or photodynamic therapy. Flagellin could be engineered to built-in adjuvanted vaccines. The built-in adjuvanted vaccines could be further engineered to target antigen presenting cells and enhance cross-presentation resulting in stronger cellular immune responses. We also improved clinical applicability by deimmunization of FlaB employing molecular dynamic analysis.