Molecular reporters to detect early stage of Avian Influenza Virus Infection: Research Team Led by Professor Jun-Seok Lee at Korea University College of Medicine Develops Fluorophore that Can Detect Avian Influenza-infected Cells

- Detection method for single cells in the early stages of infection of the avian influenza virus using new “T-shaped” fluorogenic molecules sensitive to environmental changes
- Published in Nature Communications, a world-class international academic journal

(From the left) Professor Jun-Seok Lee of Korea University, Professor Dongwhan Lee of Seoul National University, Dr. Mamunul Haque of Korea University, and Dr. Taewon Kang of Seoul National University

A stress-responsive molecular probe has been developed, which could differentiate cellular states for early diagnosis of avian influenza infection.

A joint study team led by Jun-Seok Lee, professor of Pharmacology at the KUCM and professor Dongwhan Lee of Chemistry at the Seoul National University developed new “T-shaped” fluorogenic molecules (extended and ligating imidazolyl fluorophore, EliF) and identified the molecule that selectively responds to minute changes within the endoplasmic reticulum. This molecule demonstrated light-emitting properties selectively in the cells infected with the avian influenza virus at the early stage of infection. 

Existing studies on the diagnosis of avian influenza infection have been confined to either using the PCR technique specific to the gene sequence of the virus or the detection technique of using antibodies to the avian influenza virus protein. They make direct observation of the virus regardless of the subject being infected, whereas the research team launched the study with a focus on the interaction between the virus and the host cell.

Viruses multiply actively interacting with host cells, and they have different infection sensitivities depending on the genetic and developmental characteristics of the host cells. The research team reported that the patterns of avian influenza infection are different in cell line models with different organ-derived cells and genetically different backgrounds, and based on this, new molecular probes and chemical protein analysis methods have been studied to understand the interaction between viruses and host cells at the molecular level.

The research team designed a T-shaped EliF fluorophore that sensitively responds to changes in the surrounding environment by controlling the structural flexibility of the fluorophore and built a library of similar structures. Surprisingly, uorophores localize selectively at the endoplasmic reticulum in living cells. In general, a drug linked with a fluorophore can target intracellular organelles, but properties of fluorophore have never been reported to target the endoplasmic reticulum.

Furthermore, chemical proteomics profiling and bioinformatics analysis were performed to identify proteins interacting with the EliF within cells, and it was found that proteins and EliF molecules associated with endoplasmic reticulum stress interact selectively. The endoplasmic reticulum is an organ where protein synthesis takes place, and in neurodegenerative diseases, diabetes, and viral infections, a physiological response called endoplasmic reticulum stress occurs. This means that aggregation and accumulation of abnormal folding proteins occur in the endoplasmic reticulum. Flow cytometry analysis demonstrated that EliF molecules can measure the initial change in avian influenza virus infection at the cell level as a change in fluorescence. This new analysis technology will allow researchers to study the characteristics of host cells vulnerable to infection since cells infected with the avian influenza virus can be detected and separated from complex cells and tissues.

In addition, unlike the existing infection diagnosis approach, which takes several days to complete, the method developed by the research team is significant in that the degree of infection can be quantified within 24 hours based on the intensity of fluorescence at the single cell level.

Professor Lee, who led the research said, "The EliF molecule allows us to monitor virus infection at the single-cell level in the early stage of avian influenza virus infection. This will lay the foundation on which we can determine the mechanism of action of infection and develop therapies."

The study was led by first co-authors: Dr. Taewon Kang of Seoul National University; Dr. Mamunul Haque of Korean University; and corresponding authors of Jun-Seok Lee of Korean University; and Dongwhan Lee from the Department of Chemistry at Seoul National University. The effort was supported by the Ministry of Science and ICT, Korea Biomedical technology development project (for the future infectious disease technologies) of the National Research Foundation, Medium-sized Research Support Project, and the Samsung Future Technology Promotion Program. The study entitled “Orthogonally-tunable and ER-targeting Fluorophores Detect Avian Influenza Virus Early Infection” was published on-line in the sister journal of Nature, Nature Communications (IF=17.69)>.

Research Team Led by Professor Kihoon Han from the Department of Neuroscience Identified a Novel Cause of West Syndrome, a Rare Neurodevelopmental Disorder

- Major symptoms such as spasm-like movements, microcephaly, and developmental regression were confirmed in a mouse model carrying the human genetic variant
- The study was published in an international academic journal, Annals of Neurology

 (from the left) Eunjoon Kim, the head of the Institute for Basic Science Center for Synaptic Brain Dysfunctions; Muwon Kang, a graduate student; professor Kihoon Han from the Department of Neuroscience at KUCM; Dr. Yinhua Zhang; and Hyae Rim Kang, a graduate student  

A new cause of West syndrome, a rare neurodevelopmental disorder, has been identified, inspiring hope for treatment.

A joint research team led by Professor Kihoon Han from the Department of Neuroscience at KUCM working together with another team from the Institute for Basic Science (IBS) Center for Synaptic Brain Dysfunctions (Director Eunjoon Kim) confirmed in an animal model that a single-nucleotide mutation in the CYFIP2 gene could be the cause of West syndrome.

They confirmed that the mice with CYFIP2 p.Arg87Cys variant recapitulated representative symptoms seen in patients with West syndrome, such as spasm-like movements, microcephaly, and developmental regression at an early age. It was also found that neurons were lost in the hippocampus during adulthood, and there was an overgrowth of astrocytes and microglia (gliosis). Furthermore, in terms of a molecular mechanism, it was found that the p.Arg87Cys mutation promoted ubiquitination and proteasomal degradation of the CYFIP2 protein.

Professor Han, the principal investigator, said, "The most important achievement is that this research verified the causal relationship between the CYFIP2 gene mutation and West syndrome, and specifically explained the effect of the mutation on CYFIP2 protein. The CYFIP2 mutant animal model produced for this research is of great value as it can be used to understand the detailed pathophysiological mechanism of West syndrome and to develop new therapies."

The study was led by co-first authors Muwon Kang, a graduate student at KAIST; Dr. Yinhua Zhang of KUCM; Hyae Rim Kang, a graduate student at KUCM; co-corresponding authors director Eunjoon Kim from IBS Center for Synaptic Brain Dysfunctions; and Professor Kihoon Han at KUCM. Support for the study was provided by the Ministry of Science and ICT, the National Research Foundation of Korea and the IBS Center for Synaptic Brain Dysfunctions. The study results, entitled "CYFIP2 p.Arg87Cys Causes Neurological Defects and Degradation of CYFIP2," was published in the online edition of the renowned international journal Annals of Neurology (IF=11.274) on October 17.

Students at Korea University College of Medicine Publish a Research Paper in an International Journal on AI-based Coronary Artery Calcification Scoring

- AI software measuring coronary calcium scores in chest CT will help to predict and prevent cardiovascular diseases.
- The immersive scientific research practice program leads to SCI-level paper published.

A paper written by undergraduate students at Korea University College of Medicine (KUCM) on monitoring coronary artery calcification by using artificial intelligence (AI) was published in one of the SCI-level international journals.

In February, KUCM seniors Hyun Woo Kang and Woo Jin Ahn joined the immersive scientific research practice program, implemented to encourage students to take control of their learning and to practice critical thinking. They soon were engaged in a study to evaluate the accuracy of coronary  calcium scoring (CAC) on AI-based non-ECG-gated low-dose chest computed tomography (CT) scans with different slice thicknesses under the leadership of Professor Cherry Kim from the Department of Radiology at KUCM Ansan Hospital.

CAC has been shown to be especially beneficial for predicting the occurrence of cardiovascular disease and determining a prognosis of conditions in the asymptomatic population. Up to now CAC has been measured using the ECG-gated cardiac CT scans. The research team evaluated the presence or absence and its degree of coronary artery calcification on non-ECG-gated low-dose chest CT scans using AI-based software. The research found a high level of agreement between CAC measured with the low-dose chest computed tomography screenings using AI-based software and the outcome of the conventional heart CT scans. In addition, the LDCT screening with a 1.0-mm slice thickness yielded more accurate CAC  than the LDCT with a 2.5-mm slice thickness did, particularly in high-risk patients. Reconstruction of low-dose chest CT images with a 1.0-mm slice thickness had a smaller number of underestimated CAC than did those with an image slice thickness of 2.5-mm.

The research team emphasized that AI-based software can accurately and efficiently calculate CAC based on the low-dose chest CT images taken for other purposes, and that it can be used to predict and help prevent cardiovascular diseases. In particular, those high-risk people who require lung cancer screening and the population group at high risk of cardiovascular disease share common risk factors, and in Korea, lung cancer screening is usually conducted using the low-dose chest CT scan, which makes the research outcome more useful.

Co-authors Kang and Ahn said, "We are really grateful that Professor Kim led us in performing such good research through the immersive science practice program, and we are delighted the results of our efforts have finally been published. The CAC can help predict the occurrence of cardiovascular diseases and help us determine treatment approaches. The conventional measurement approach has been labor-intensive and time-consuming, so we expect our AI-based assessment can help both the doctors and patients with its greater efficiency."

Professor Kim, the research team leader, said, "We were researching together as a team; I was not just giving them instructions. Rather, we had many discussions whenever we were faced with challenges while working on this project. I feel honored to have had this opportunity of working with two such great students. It is meaningful in that this research lays the foundation on which future studies can be done to evaluate coronary artery calcium scores using low-dose chest CT scans."

The paper “Evaluation of Fully Automated Commercial Software for Agatston Calcium Scoring on Non-ECG-gated Low-dose Chest CT Scans with Different Slice Thicknesses” was published in the latest issue of the international journal, European Radiology (IF: 7.034).

A New Zoonotic Virus Discovered in China is Similar to the Virus Found in Korea Last Year
- A team led by Jin-Won Song of Korea University and Won-Keun Kim of Hallym University revealed that the Gamak Virus and Daeryong Virus found in Korea are similar to the Langya henipavirus of China
-As it infects humans, we need to prepare for the emergence of a new virus

A zoonotic virus recently discovered in Shandong and other Chinese provinces is of the same genus as the ones discovered in Korea last year, according to a Korean research team.

Professor Jin-Won Song's team from the Department of Microbiology at Korea University College of Medicine, and Professor Won-Keun Kim's team from Hallym University College of Medicine were the first to discover four new paramyxoviruses, including one in the Henipavirus family, in three species of rodents and carnivorous animals living in South Korea last year. The discovery was published in Virology and Viruses, well-known international academic journals.

Prof. Song's team discovered novel paramyxoviruses in the insectivorous animals known to have hantavirus such as the Ussuri shrew and the small shrew, and named them the Gamak and Daeryong viruses respectively. These two viruses were recently discovered in the same host animal where the new Langya henipavirus from Shandong Province, China, was found as published in the international academic journal, The New England Journal of Medicine, by Chinese and Singaporean researchers. And their similarity of RNA put them in the same genus.

Henipavirus, transmitted from fruit bats that inhabit in Southeast Asia, is known to have a very high (up to 70%) fatality rate, although the transmissibility is not great. On the other hand, it was announced that the Langya virus found in China would not cause fatal or very serious diseases.

Professor Jin-Won Song from the Department of Microbiology said, “Judging from the Langya virus infection cases up to now, it seems that Gamak and Daeryong viruses found in Korea are not fatal to humans, either. However, those infected will have symptoms such as fever, fatigue, cough, and pneumonia, so additional research should be conducted to brace for the emergence of a new virus.”