암 관련 비코딩 RNA에 의해 형성되는 분자 구조물의 기능적 분할 분석

학위논문(박사) - 한국과학기술원 : 화학과, 2013.8 ,[ix, 119 p. :]Recent studies have shown that RNAs act as active molecules in the regulation of various cellular metabolism processes and plays a key role in cancer development or other disease progression. Their biological roles depend on the three-dimensional structures, although specific sequences have been shown to be crucial for various functions. Therefore, analysis of the RNA conformation in cells is essential for understanding their roles. First, we devised a novel tool for detecting and analyzing RNA conformation by anti-RNA antibody. We have developed an efficient strategy for panning and affinity maturation of human monoclonal antibodies binding to RNA from a na"ive antigen binding fragment (Fab) combinatorial phage library, using brain cytoplasmic 200 (BC200) RNA as the antigen. BC200 RNA, a neuron-specific noncoding RNA, is implicated in the inhibition of local synaptodendritic protein synthesis in human cells, and shown to be expressed at higher levels in invasive carcinomas than benign tumors of the breast. Using panning and affinity maturation, we identified MabBC200-A3 as the optimal binder, which interacted specifically with two regions (residues 76 to 85 and 96 to 104) of BC200 RNA with a dissociation constant of ~7 nM. Expression of BC200 RNA in various breast cancer cell lines was further examined using conventional hybridization and immunoanalysis with MabBC200-A3. When total cellular RNAs purified from cells were subjected to hybridization, the signals varied significantly among the cell types. Similar variability of BC200 levels among cell lines was observed upon immunoanalysis with the antibody. Furthermore, the antibody was able to discriminate BC200 RNA from homologous 7SL RNA, supporting its utility as a specific probe for the RNA. Intriguingly, however, when permeabilized cells were used instead of purified total cellular RNA, the amounts of antibody-recognizable BC200 RNA were different, indicating that BC200 RNA exists as at least two distinct cellular forms (antibody-recognizable and nonrecognizable), depending on the structural integrity of the antigenic RNA motif and its accessibility to the antibody. Our data clearly demonstrate the value of the anti-RNA antibody as a novel tool for investigating RNA conformation, which cannot be achieved with hybridization. Second, we analyzed the structural elements of steroid receptor RNA activator (SRA) RNA and suggested a new RNA inhibitor on estrogen receptor alpha (ER $\alpha$) signaling in breast cancers. SRA is an RNA regulator that coactivates steroid receptor-mediated transcription. It is a putative coactivator in the estrogen receptor (ER) signaling pathway. It functions as an RNA molecule itself, but its isoform can encode a protein. SRA levels are elevated in the majority of tumors. Regulatory actions of noncoding SRA core RNA vary depending on the type of estrogen receptors, the presence or absence of ligand, and even reporter elements. Recently it was reported that SRA RNA can be dissected into four domains based on various probing experiments. In this study, we examined the contribution of domains or helices to its coactivation ability to understand the role of structural elements of SRA RNA essential for the RNA function. We showed that domain III played a crucial role in SRA RNA. Furthermore the domain specifically inhibits ER $\alpha$ -dependent transactivation as a competitive inhibitor by interfering with the interaction between coactivators and SRA RNA. In particular four helices, H15 to H18 among nine helices of the domain III showing significantly high sequence conservation across vertebrates also inhibited the ER $\alpha$ signaling pathway. Taken together, the results gave an insight into structural basis for a role of SRA RNA with the estrogen and ER complex and proposed a new target candidate on RNA therapeutics in breast cancers. In summary, this thesis proposed a novel tool for recognizing RNA conformation and dissecting RNA structural motifs. Also we investigated the contribution of structural elements of SRA RNA in breast tumors. In that molecular architecture plays a key role in understanding the mechanism of many functional RNAs, these studies of functional dissection can pave the way for gaining extensive insights on RNA field as well as application to develop new diagnostic and therapeutic approaches for RNA-related diseases.한국과학기술원 :화학과

A Study on the Outlet Blockage Determination Technology of Conveyor System using Deep Learning

본 연구는 컨베이어 시스템에서 딥러닝을 이용한 배출구 막힘 판단 기술에 대하여 제안한다. 제안 방법은 산업 현장의 CCTV에서 수집한 영상을 이용하여 배출구 막힘 판단을 위한 다양한 CNN 모델들을 학습시키고, 성능이 가장 좋은 모델을 사용하여 실제 공정에 적용하는 것을 목적으로 한다. CNN 모델로는 잘 알려진 VGGNet, ResNet, DenseNet, 그리고 NASNet을 사용하였으며, 모델 학습과 성능 테스트를 위하여 CCTV에서 수집한 18,000장의 영상을 이용하였다. 다양한 모델에 대한 실험 결과, VGGNet은 99.89%의 정확도와 29.05ms의 처리 시간으로 가장 좋은 성능을 보였으며, 이로부터 배출구 막힘 판단 문제에 VGGNet이 가장 적합함을 확인하였다.22Nkc

A PoD(Probability of Damage) Prediction Monitoring System for Structure Safety based-on Deep Learning

최근 경주와 포항의 지진 발생 이후, 구조물 노후화 정도와 구조물 붕괴 위험도의 사전 탐지 및 피해 대응을 위한 구조물 건전성 모니터링(SHM, Structural Health Monitoring) 기술이 주목을 받고 있다. 본 논문에서는 구조물의 센서 데이터와 1-D CNN 모델을 이용하여 구조물 건전성 모니터링을 수행하고, 구조물 건전성 모니터링에 IoT를 결합하여 딥러닝 기반 구조물 손상확률(PoD, Probability of Damage) 예측 시스템을 제안한다. 1-D CNN 모델을 활용한 구조물 손상확률 예측 모델에서는 구조물 센서 데이터를 프레임 단위로 나누어 예측 속도를 빠르게 하였고, 전체 프레임들의 손상확률 평균으로 결과를 산출하여 예측 정확도를 개선하였다. 이를 활용하여, 제안한 시스템은 구조물 유지 보수에 대한 의사결정 지원 및 사전 경보 등을 통하여 지진과 같은 재난으로부터 발생하는 피해를 최소화하는 분야에 적용할 수 있다.2

A pharmaceutical composition for inhibiting metastasis comprising siRNA of BC200 RNA

본 발명은 BC200 RNA의 활성을 억제할 수 있는 siRNA, 상기 siRNA로 세포내에서 변환될 수 있는 shRNA를 발현시키는 벡터 및 상기 siRNA 또는 벡터를 포함하는 종양전이 억제용 약학조성물에 관한 것이다. 본 발명의 BC200 RNA에 대한 siRNA는 BC200 RNA의 활성을 억제하여, 암세포의 이동 및 이에 따른 암세포의 전이를 억제할 수 있으므로, 보다 효과적인 암의 치료에 널리 활용될 수 있을 것이다

RNA-특정 결합된 항체

The present invention relates to an antibody being capable of binding specifically to BC200 RNA or its antigen binding fragment, a polynucleotide of encoding the antibody or its antigen binding fragment, a composition, kit and method for detecting BC200 RNA in a sample using the antibody or its antigen binding fragment, and a composition for diagnosis, prevention or treatment of BC200 RNA-associated diseases including the antibody or its antigen binding fragment

RNA-SPECIFIC BINDING ANTIBODY AND SELECTING METHOD FOR THE SAME

본 발명은 RNA에 결합하는 항체의 선별방법 및 BC200 RNA에 특이적으로 결합하는 항체에 관한 것이다. 본 발명의 선별방법은 기능성 RNA의 구조를 인식하는 인간 항체의 제조에 유용하게 사용되며, BC200 RNA에 특이적으로 결합하는 항체는 BC200 RNA를 인지하거나 그 작용을 조절할 수 있어 관련 의약분야에 다양하게 적용될 수 있다