Modular protein-DNA hybrid nanostructures as a drug delivery platform

Ryu, Yiseul,Hong, Cheol Am,Song, Yunjin,Beak, Jonghwi,Seo, Bo Am,Lee, Joong-jae,Kim, Hak-Sung The Royal Society of Chemistry 2020 Nanoscale Vol.12 No.8

<P>With the increasing number of identified intracellular drug targets, cytosolic drug delivery has gained much attention. Despite advances in synthetic drug carriers, however, construction of homogeneous and biocompatible nanostructures in a controllable manner still remains a challenge in a translational medicine. Herein, we present the modular design and assembly of functional DNA nanostructures through sequence-specific interactions between zinc-finger proteins (ZnFs) and DNA as a cytosolic drug delivery platform. Three kinds of DNA-binding ZnF domains were genetically fused to various proteins with different biological roles, including targeting moiety, molecular probe, and therapeutic cargo. The engineered ZnFs were employed as distinct functional modules, and incorporated into a designed ZnF-binding sequence of a Y-shaped DNA origami (Y-DNA). The resulting functional Y-DNA nanostructures (FYDN) showed self-assembled superstructures with homogeneous morphology, strong resistance to exonuclease activity and multi-modality. We demonstrated the general utility of our approach by showing efficient cytosolic delivery of PTEN tumour suppressor protein to rescue unregulated kinase signaling in cancer cells with negligible nonspecific cytotoxicity.</P>

Increase in the Detection Sensitivity of a Lateral Flow Assay by Oriented Immobilization of Antibody

Yiseul RYU,Zongwen JIN,Mun Suk KANG,Hak-Sung KIM 한국생물공학회 2012 한국생물공학회 학술대회 Vol.2012 No.4

Development of a Highly Sensitive Phage-Based Biosensor for Cancer Diagnosis

JuHyun BAE,YiSeul RYU,SeongJoon KIM 한국생물공학회 2022 한국생물공학회 학술대회 Vol.2022 No.9

온라인 학습카페 활용 수업에 대한 학생평가 및 학습활동 유형 분석 : S대학교 WIU 강좌 사례를 중심으로

김경은(Kim Kyoungeun),장윤호(Jang YunHo),류이슬(Ryu Yiseul) 한국어린이미디어학회 2020 어린이미디어연구 Vol.19 No.4

최근 교육환경의 변화 등으로 다양한 형태의 온라인 기반 학습이 이루어지고 있다. 본 연구는 포털사이트 기반 학습카페를 활용한 수업에 대한 학생평가 및 구체적인 학습활동 유형 분석을 통해 온라인 기반 수업의 개선방안을 탐색하고자 한다. 이를 위해 2019년 S대학교에서 운영된 65개 온라인 학습카페 활용 강좌를 대상으로 수업효과에 대한 학생평가 설문을 실시하였다. 더불어 수업별 운영된 학습활동을 학습형태, 활동수준, 상호작용 방식을 중심으로 유형화하여 분석하였다. 분석 결과, 학생들의 수업에 대한 만족도는 3.73점으로 나타났고, 인지적 차원의 수업효과에 대한 인식이 3.84점으로 가장 높게 나타났다. 한편, 학습활동의 유형은 개인기반/기억 및 이해 수준의 활동/간접적 상호작용 방식이 38.46%로 가장 높은 비율을 보였고, 개인기반/적용 및 분석 수준의 활동/간접적 상호작용 방식이 26.15%로 높게 나타났다. 이는 수업에서 온라인 학습카페가 ‘개인의 학습활동 결과를 공유하는 장’으로 주로 활용되고 있음을 의미한다. 이러한 분석결과를 토대로 보다 나은 수업효과를 위한 온라인 학습카페 활용을 위한 논의점을 제시하였다. The purpose of this study is to explore direction for improvement of courses using a web-based bulletin board in college education from course evaluation of students and analysis of learning activity types. To achieve this aim, a survey of course evaluation was conducted on students in 65 courses which used a web-based bulletin board at S University in 2019. And learning activities performed in these courses were analyzed by elements such as the type of learning(individual based/team based), the level of cognition(remember/understand/apply/analyze/evaluate/create) and the type of interaction (direct interaction between professor and student/direct interaction between student and student/indirect interaction). The results of the study are as follows: first, students’ satisfaction with the courses was showed 3.73 points and students’ awareness of cognitive effects was showed 3.84 points. Second, the type of learning activities asked that individuals usually carried out tasks for knowledge and comprehension without direct interaction between professor and students was showed 38.46%. And the type of learning activities asked that individuals usually carried out tasks for application and analysis without direct interaction between professor and students was showed 26.15%. These findings mean that a web-based bulletin board were mainly used as a venue for sharing the result of learning activities by individual rather than producing new knowledge based on cooperation in courses.

벼메뚜기(Oxya chinensis sinuosa Mistshenko) 분말 섭취와 유산소성 운동훈련에 의한 마우스의 에너지 대사 변화

김이슬,전병덕,최석립,김우철,이동운,류승필,Kim, Yiseul,Jeon, Byungduk,Choi, Seokrip,Kim, Woocheol,Lee, Dong Woon,Ryu, Sungpil 한국응용곤충학회 2016 한국응용곤충학회지 Vol.55 No.1

본 연구의 목적은 벼메뚜기(Oxya chinensis sinuosa) 분말 섭취가 유산소성 운동훈련(트레드밀 달리기)의 병행 유무에 의해 ICR 생쥐의 에너지 대사를 증가시키는지를 알아보고자 하는 것이다. 이 목적을 달성하기 위하여, 28 마리의 ICR 생쥐를 보통식 대조군(CON), 보통식 대조군으로서 운동훈련 병행군(COEX), 벼메뚜기 분말이 혼합된 사료 섭취군(GH), 그리고 벼메뚜기 분말이 혼합된 사료를 섭취함과 동시에 운동훈련을 병행군(GHEX)으로 구분하였다. 벼메뚜기 분말 사료섭취 및 운동은 6주간 진행하였다. 체중증가율은 유의하지 않았다. 지방량은 GH와 GHEX에서 유의하게 감소하였다. 혈중 glutamic oxaloacetic transaminase와 glutamic pyruvic transaminase 수준은 처치 집단간 변화가 없었다. 제2형 당수송체 및 제4형 당수송체는 처치 집단간 유의한 차이가 없었다. GHEX의 fibronectin type III domain-containing protein 5 단백질 발현량이 가장 높았다. AMP-activated protein kinase 단백질 수준은 GHEX에서 유의하게 증가하였다. Glycogen synthase kinase 3 beta 단백질 발현량은 GHEX가 CON과 비교할 때 감소하였다. 이러한 결과들은 벼메뚜기 분말을 섭취하면서 지구성 운동훈련을 하는 경우에 에너지 대사에 영향을 준다는 것을 제시하고 있다. The purpose of this study was to analyze the effect of grasshopper (Oxya chinensis sinuosa) powder ingestion with/without aerobic exercise (treadmill running) on energy metabolism. To achieve this purpose, 28 Institute of Cancer Research (ICR) mice were divided into four groups: normal diet control group (CON), a normal diet with exercise control group (COEX), a grasshopper powder-supplemented diet group (GH), and a grasshopper powder-supplemented diet with exercise group (GHEX). Duration of the powder ingestion and aerobic exercise training were 6 weeks. Body weight gain ratio was not significant. Fat mass significantly decreased in GH and GHEX. There were no changes in blood glutamic oxaloacetic transaminase and glutamic pyruvic transaminase levels between groups. Glucose transporter type 2 and glucose transporter type 4 protein levels were not significantly different between groups. Fibronectin type III domain-containing protein 5 level was the highest in GHEX. AMP-activated protein kinase level significantly increased in GHEX compared to the levels in the other groups. Glycogen synthase kinase 3 beta protein level was reduced in GHEX compared to that in CON. These results suggest that grasshopper powder ingestion and endurance exercise training influence energy metabolism.

누에나방(Bombyx mori L.) 번데기 분말 섭취가 마우스의 저항성운동 훈련 후 근육량 증가에 미치는 영향

김이슬,김희빈,전병덕,이동운,류승필,Kim, Yiseul,Kim, Heebin,Jeon, Byungduk,Lee, Dong Woon,Ryu, Sungpil 한국응용곤충학회 2016 한국응용곤충학회지 Vol.55 No.1

식용곤충은 대체 식량자원으로 관심이 증가되고 있다. 본 연구의 목적은 누에나방 번데기 분말 섭취 유무에 따른 저항성운동 훈련(등장성 수축)이 ICR 마우스의 근육량 증가에 미치는 영향을 분석하였다. 이를 위하여 ICR 마우스 28마리를 대조군(CON), 저항성운동 훈련군(EX), 누에나방 번데기 분말 섭취군(SP), 누에나방 번데기분말 섭취 저항성운동 훈련군(SPEX)으로 그룹 당 각각 7마리씩 분류하였다. 체중증가율은 EX와 SPEX가 CON과 SP에 비해 유의하게 억제되었다. 혈중 총 단백질 농도는 SPEX가 다른 그룹에 비하여 가장 높았다. 알부민 농도 변화는 운동훈련 병행 시에만 증가하였다. 혈액 GOT와 GPT 수준은 유의차가 없었다. Akt와 Gsk-$3{\beta}$의 단백질 발현의 유의차는 없었다. 그러나 번데기 분말 섭취 시 Akt 증가 및 Gsk-$3{\beta}$의 감소 경향이 나타났다. 비복근 근육 양은 저항성 운동 훈련을 하였을 때만 증가하였다. 그리고 또한, 번데기 분말 섭취 시 근육량은 유의차가 없었으나 증가하는 경향을 보였다. 이러한 결과는 저항성 운동훈련과 누에나방 번데기 분말 섭취는 간독성을 유발하지 않고 근육량을 증가시킬 수 있다는 점을 시사한다. 그러나 보다 상세한 결과를 제시하기 위해 부가적인 연구가 필요할 것이다. Edible insects are an interesting alternative global food resource. The purpose of this study was to investigate whether ingestion of silkworm (Bombyx mori) pupae powder with and without resistance exercise training (isometric contraction training) increased muscle mass in ICR mice. To achieve this, 28 ICR mice were grouped into control (CON), resistance exercise training (EX), silkworm pupae powder ingested control (SP), and silkworm powder ingestion with resistance exercise training (SPEX) groups. The change in body weight ratio was significantly decreased in the EX and SPEX groups compared to the CON and SP groups. Total blood protein levels were the highest in SPEX mice compared to those in other groups. The albumin concentration increased only in the EX group. Blood GOT and GPT levels were not significantly affected. Changes in Akt and Gsk-$3{\beta}$ protein expression were not significant but there was a tendency for Akt to increase and for Gsk-$3{\beta}$ to increase following the ingestion of the powder. The size of the gastrocnemius muscle increased significantly in response to resistance exercise training only. Furthermore, the ingestion of silkworm pupae powder tended to increase muscle mass without significance. These results suggested that the ingestion of silkworm pupae powder with resistance exercise training might enhance muscle mass without hepatotoxicity. However, future study may be needed to obtained detailed results and practical suggestions.

Genetically engineered and self-assembled oncolytic protein nanoparticles for targeted cancer therapy

Lee, Joong-jae,Kang, Jung Ae,Ryu, Yiseul,Han, Sang-Soo,Nam, You Ree,Rho, Jong Kook,Choi, Dae Seong,Kang, Sun-Woong,Lee, Dong-Eun,Kim, Hak-Sung IPC Science and Technology Press 2017 Biomaterials Vol.120 No.-

<P><B>Abstract</B></P> <P>The integration of a targeted delivery with a tumour-selective agent has been considered an ideal platform for achieving high therapeutic efficacy and negligible side effects in cancer therapy. Here, we present engineered protein nanoparticles comprising a tumour-selective oncolytic protein and a targeting moiety as a new format for the targeted cancer therapy. Apoptin from chicken anaemia virus (CAV) was used as a tumour-selective apoptotic protein. An EGFR-specific repebody, which is composed of LRR (Leucine-rich repeat) modules, was employed to play a dual role as a tumour-targeting moiety and a fusion partner for producing apoptin nanoparticles in <I>E. coli</I>, respectively. The repebody was genetically fused to apoptin, and the resulting fusion protein was shown to self-assemble into supramolecular repebody-apoptin nanoparticles with high homogeneity and stability as a soluble form when expressed in <I>E. coli</I>. The repebody-apoptin nanoparticles showed a remarkable anti-tumour activity with negligible side effects in xenograft mice through a cooperative action of the two protein components with distinct functional roles. The repebody-apoptin nanoparticles can be developed as a systemic injectable and tumour-selective therapeutic protein for targeted cancer treatment.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Amelioration of DSS-Induced Acute Colitis in Mice by Recombinant Monomeric Human Interleukin-22

Kim Suhyun,Hong Eun-Hye,Lee Cheol-Ki,Ryu Yiseul,Jeong Hyunjin,Heo Seungnyeong,이중재,고현정 대한면역학회 2022 Immune Network Vol.22 No.3

IL-22, a pleiotropic cytokine, is known to have a profound effect on the regeneration of damaged intestinal barriers. The tissue-protective properties of IL-22 are expected to be potentially exploited in the attenuation and treatment of colitis. However, because of the disease-promoting role of IL-22 in chronic inflammation, a comprehensive evaluation is required to translate IL-22 into the clinical domain. Here, we present the effective production of soluble human IL-22 in bacteria to prove whether recombinant IL-22 has the ability to ameliorate colitis and inflammation. IL-22 was expressed in the form of a biologically active monomer and non-functional oligomers. Monomeric IL-22 (mIL-22) was highly purified through a series of 3 separate chromatographic methods and an enzymatic reaction. We reveal that the resulting mIL-22 is correctly folded and is able to phosphorylate STAT3 in HT-29 cells. Subsequently, we demonstrate that mIL-22 enables the attenuation of dextran sodium sulfate-induced acute colitis in mice, as well as the suppression of pro-inflammatory cytokine production. Collectively, our results suggest that the recombinant mIL-22 is suitable to study the biological roles of endogenous IL-22 in immune responses and can be developed as a biological agent associated with inflammatory disorders.

Enzymatic Prenylation and Oxime Ligation for the Synthesis of Stable and Homogeneous Protein–Drug Conjugates for Targeted Therapy

Lee, Joong‐,jae,Choi, Hyo‐,Jung,Yun, Misun,Kang, YingJin,Jung, Ji‐,Eun,Ryu, Yiseul,Kim, Tae Yoon,Cha, Young‐,je,Cho, Hyun‐,Soo,Min, Jung‐,Joon,Chung, Chul‐,Wo WILEY‐VCH Verlag 2015 Angewandte Chemie Vol.127 No.41

<P><B>Abstract</B></P><P>Targeted therapy based on protein–drug conjugates has attracted significant attention owing to its high efficacy and low side effects. However, efficient and stable drug conjugation to a protein binder remains a challenge. Herein, a chemoenzymatic method to generate highly stable and homogenous drug conjugates with high efficiency is presented. The approach comprises the insertion of the CaaX sequence at the C‐terminal end of the protein binder, prenylation using farnesyltransferase, and drug conjugation through an oxime ligation reaction. MMAF and an EGFR‐specific repebody are used as the antitumor agent and protein binder, respectively. The method enables the precisely controlled synthesis of repebody–drug conjugates with high yield and homogeneity. The utility of this approach is illustrated by the notable stability of the repebody–drug conjugates in human plasma, negligible off‐target effects, and a remarkable antitumor activity in vivo. The present method can be widely used for generating highly homogeneous and stable PDCs for targeted therapy.</P>