(A)Study on Printed Polydiacetylene Vesicles as Chemical Sensors

이승우 고려대학교 대학원 2009 국내석사

There has been a great interest on mimicking the structure of cell membranes for their unique properties from a material science point of view. The surface of a cell membrane contains highly specific molecular recognition receptor sites and forms mosaics which has mobility. One of functional monomors such as diacetylene molecules can form a membrane-like structure by self-organization and functionalization. Vesicles and films can be formed by Self-organization of diacetylene monomers into supramolecular assemblies and they can be polymerized by irradiation with 254 nm UV light. These supramolecules undergo a visible color transition from blue by polymerization to red due to various environmental stimuli including temperature, pH, chemical solvent, ligand-receptor interaction and mechanical stress. The color transition occurs when external stimuli impose stresses altering delocalization length of π electrons along polydiacetylene (PDA) backbones. Recently, our group were successful in mixing different diacetylene monomers and preparing vesicles for immobilization of the polydiacetylene liposomes on glass, ZnO, SiC and GaN substrates without losing their unique color changing property. However, we mixed monomers for the purpose of immobilization of vesicles. In this study, we mixed two diacetylene monomers having two different functional group (carboxylic and amine groups) and showed its colorimetric transition by different mixing molar ratio and how it effects the sensitivity of vesicles compared to homogeneous vesicles. We also test for the adjustment of an inkjet printer for printing polydiacetylene vesicles. Optimal condition of inkjet printer for PDA system was found. We report on patterning on various substrates using inkjet printer to make patterns more recognizable such as letters. Each letter is found to possess the color-changing property as well as the fluorescence emission. We have developed a new approach for the construction of label-free PDA sensor which is compatible with conventional inkjet technologies. This sensor fabricated by an inkjet printer was applied to a chemical sensor. By using specific ligand-receptor interaction occurring between cyclodextrins and PDA vesicles and ionic interaction occurring between acid group and amine group of gas analytes, printed letters with fluorescence profiles were generated. In this experiment we have investigated printed images of PDA vesicles integrated on flexible substrates such as nitrocellulose membrane and glass fibers via an inkjet printer. The shape and size of printed PDA vesicles on all the substrates were uniform and reproducible. The size of them was various from 0.5 cm to 1 cm for a letter. This detection method could be applied as DNA chip, protein chip, and cell chip for multiple screening as well as wall paper-like flexible chemical sensors by modifying the functional groups of diacetylene monomer.

Sensitivity enhancement of polydiacetylene vesicles by optimizing vesicles size and polymerization temperature

오재호 고려대학교 대학원 2009 국내석사

Diacetylene-funtionalized lipids can be used as the fundamental materials of effective bio-chemical sensors. Polydiacetylene vesicles change visible color from blue to red under external stimuli such as temperature, solvent, pH, mechanical stress and α-cyclodextrin. The color transition occurs when external stimuli impose stresses altering delocalization length of π-electrons in the repeated ene-yne bond moieties along polydiacetylene backbones. Most of the polydiacetylenes are prepared by 254-nm UV light of self-assembled monomers in vesicles solutions or Langmuir-Blodgett/Langmuir-Schaefer films on solid substrates. We previously investigated cyclodextrin-induced color changes in a polymerized diacetylene Langmuir-Schaefer film or vesicle solution. Cyclodextrins are well known for their ability to form inclusion complexes with various substrates. It would be useful to develop more understanding and design of PDA based chemosensors. Meantime a variety of methods to increase the sensitivity in polydiacetylene systems like variation of carbon chain length, polymerization temperature and strength of UV irradiation were reported in previous papers. Recently, it was published that smaller the vesicles are, the more sensitive is in detecting concanavalin (Con A) when polydiacetylene vesicles incorporated with glycolipids. In this study we investigated the effect of sensitivity enhancement polydiacetylene vesicles in detection α-cyclodextrin according to vesicle size by filtration technology with different pore sizes and process with different polymerization temperature. The degree of color change is quantified by UV-vis spectrometer. In order to quantify the response of polydiacetylene vesicles to the α-cyclodextrin, it can be defined colorimetric response that represents degree of color change from blue to red.

식물세포에서 분리한 Tonoplast Vesicles에서의 Ca^(2+)수송과 Inositol 1, 4, 5-trisphosphate(IP₃)의 작용

김현주 이화여자대학교 대학원 1989 국내석사

A Study on Vesicles of Multidrug-Resistant Bacteria against Beta-lactam Antibiotics

김시원 경상대학교 대학원 2020 국내박사

항생제의 발견은 인간의 수명을 연장시켰으며 의료기술의 발전에 큰 도움을 줬다. 최초의 항생제인 페니실린의 발견 이후 여러 개량 항생제들이 만들어져 왔으나 항생제의 오남용으로 인하여 항생제에 대해 저항하는 세균들이 등장했다. 결국은 어떤 강력한 항생제에도 저항할 수 있는 세균이 생겨났으며 이를 슈퍼박테리아, superbugs, 또는 다제내성균이라고 한다. 이러한 다제내성균주의 출현은 최근 의학계뿐만 아니라 축산, 수산, 환경, 식품 등에서 가장 큰 문제점으로 나타나고 있으며 전세계의 공중 보건에 위협을 가하고 있다. 세균의 바깥부분이 주변세포질을 에워싸면서 형성된 나노 사이즈의 구형 물질을 vesicles라고 하며 그람-음성세균의 경우는 outer membrane vesicles (OMVs), 그람-양성세균은 extracellular vesicles (EVs)라고 부른다. 이들이 형성되는 과정에서 다양한 세균 유래의 단백질, 지질, 유전적 물질이 포함되며 이들이 가지는 이중막의 안전한 구조를 이용하여 다양한 물질을 분비 및 전달할 수 있는 장점을 가지고 있다. 바이오필름을 형성하여 주변미생물의 보호, 숙주세포에 독성물질 전달, 경쟁관계의 미생물 공격, 세균간의 통신, 세균의 영양분 탐식 기능 등이 있으며 최근에는 세균의 자기방어, 항생제 내성유전물질 전달, 항생제의 분해능력에 관해 활발히 연구가 늘어나고 있다. 본 연구에서는 그람-양성균을 스트레스 환경 혹은 정상환경에서 배양시켜 EVs를 분리하여 이들의 항생제 소모성 능력과 단백체를 비교분석하였다. 또한 conjugation assay를 통해 항생제 내성 대장균을 제작하였으며 이들에서 분리한 OMVs와 감수성 대장균에서 분리한 OMVs의 항생제 분해능력과 단백체 비교분석을 통해 베타-락탐 항생제 저항성 단백질을 유추하였으며 이들에 대한 유전체 결손돌연변이주를 제작하여 OMVs에서 포린과 베타-락타마제의 중요성을 입증하였다. 본 연구를 통해 세균은 vesicles을 분비하여 항생제에 저항할 수 있음을 증명하였으며 이는 세균의 첫 번째 방어기전으로 작용할 수 있다. 따라서 전 세계적으로 공중 보건에 위협을 가하고 있는 항생제 내성 문제에 대항하기 위해 본 연구는 도움을 줄 수 있을 것으로 판단된다. Chapter 1: 특정 세균 유래 물질을 갖고있는 extracellular vesicles (EVs)는 그람-양성 세균에서 자연적으로 유리된다. 분비된 EVs는 세균을 보호하는데 중요한 역할을 하며, 항생제 환경에서 세균의 생존에 기여할 수 있다. 이 연구에서, 우리는 메티실린 내성 포도상알균 (MRSA)의 배양 시 최소억제농도 (MIC)보다 낮은 용량의 ampicillin 항생제를 첨가하여 스트레스를 준 환경과 스트레스를 주지 않은 환경에서 EVs를 각각 분리했다. 우리는 각각의 EVs가 몇몇 종류의 베타-락탐 계열 항생제로부터 항생제 감수성 그람-양성균과 그람-음성균을 보호하고 항생제를 직접 분해할 수 있는지 조사했다. 두가지의 EVs에서 단백체 비교분석을 수행하여 베타-락탐 저항성 결정인자를 조사했다. Ampicillin 스트레스 상태에서 MRSA의 EVs의 분비량은 스트레스가 없는 환경에서 EVs의 분비량과 비교 시 급격하게 증가했다. 베타-락탐 항생제의 분해 능력과 관련된 단백질들은 스트레스 상태에서 분리한 EVs에서 정상환경의 EVs보다 풍부했다. 그러므로, 이들 EVs는 농도 의존적으로 베타-락탐 항생제를 분해하고 베타-락탐 항생제로부터 베타-락탐 감수성 세균의 생존을 도울 수 있다. 종합하자면, 이 연구를 통해 MRSA의 EVs가 베타-락탐 항생제에 대한 첫째 방어선 역할을 함으로써 베타-락탐 감수성 세균의 생존에 결정적인 역할을 하며, 항생제 스트레스는 더 높은 방어 능력을 갖는 EVs를 방출할 수 있다는 점을 밝혔다. Chapter 2: 다양한 세균 유래물질을 함유하는 outer membrane vesicles (OMVs)는 모든 그람-음성 세균에서 방출된다. 분비된 OMVs는 세균이 스스로를 방어할 수 있는 능력에 중요한 역할을 하며, 세균의 생존에 기여한다. 이 연구에서 우리는 conjugation assay를 통해 제작한 베타-락탐 항생제 내성 대장균과 recipient 베타-락탐 항생제 감수성 대장균에서 OMVs를 분리했으며, 단백질 비교분석을 통해 OMVs들이 베타-락탐 항생제 저항성 단백질을 갖고 있는지 조사했다. 우리는 또한 두 유형의 OMVs 모두 베타-락탐 항생제로부터 감수성 세균을 보호할 수 있는지 그리고 베타-락탐 항생제를 직접 분해할 수 있는지의 여부를 조사했다. 베타-락탐 항생제 분해에 관여하는 몇몇 단백질들은 베타-락탐 내성 대장균에서 분리한 OMVs에서 더 풍부했으며 이 OMVs는 농도에 비례하여 베타-락탐 항생제를 직접적으로 분해할 수 있었다. 이를 통해 베타-락탐 감수성 대장균과 다른 이종의 균주들을 베타-락탐 항생제로부터 구제할 수 있었다. 종합하자면, 본 연구는 베타-락탐 내성 대장균의 OMVs가 베타-락탐 항생제 감수성 세균의 생존에 중요한 역할을 함을 증명했다. 이 발견은 현제 전 세계적으로 공중 보건에 위협을 주고있는 항생제 내성 문제에 대항하기 위한 새로운 길을 열어줄 수 있을 것으로 판단된다. Chapter 3: 그람-음성 세균은 항생제의 진입을 억제하는 외막을 가지고있으며 외막에는 베타-락탐 항생제의 투과성을 조절하는 포린을 갖고있다. 세균의 주변세포질공간에 존재하는 베타-락타마제 효소는 베타-락탐 항생제의 항균 특성을 분해하여 항생제를 비활성화 시킨다. 흥미롭게도, 이들 포린 및 베타-락타마제는 베타-락탐 내성 대장균의 outer membrane vesicles (OMVs)에서 발견되며, OMVs의 베타-락탐 항생제 가수분해에 관여하여 항생제 환경에서 감수성 균주의 생존에 관여할 수 있다. 이 연구에서, 포린 또는 베타-락타마제의 결손이 OMVs의 생산량에 영향을 끼치는지 알아봤다. 그리고 결손돌연변이체와 모균주에서 분리한 OMVs를 비교하여 이들이 베타-락탐 항생제 소모성에 영향을 끼치는지 알아봤다. 베타-락타마제 결손 돌연변이주에서 분리한 OMVs는 베타-락탐 항생제 소모성 능력을 보이지 않았으며, OmpC나 OmpF 포린이 결손된 돌연변이주의 OMVs는 모균주의 OMVs보다 확실히 낮은 항생제 분해능을 나타냈다. 이러한 데이터는 OmpC 및 OmpF 단백질이 OMVs의 내강 내로 베타-락탐 항생제를 투과시키며, OMVs내로 들어간 항생제는 베타-락타마제에 의해 분해될 수 있음을 입증한다. 이 연구의 결과를 통해 세균의 방어기전에서 OMVs가 중요함을 증명하였으며, 이는 항생제 내성에 대한 세계적인 문제를 퇴치하는데 도움이 될 수 있다.

Studies on the role of extracellular vesicles in intercellular communication network

이은영 포항공과대학교 일반대학원 2012 국내박사

Communication between cells and the environment is an essential process in living organisms, and intercellular communication is believed to be mediated mainly by the secretion of soluble factors, cell-to-cell contacts, and tunneling machinery such as nanotubes. Recently, a mechanism mediated by extracellular vesicles, termed EVs, which are spherical, bilayered proteolipids with an average diameter of 0.03 to 1 m, has drawn much attention. Throughout evolution, both prokaryotic and eukaryotic cells have adapted to manipulate EVs for intercellular communication via (outer) membrane vesicles in the case of bacteria and exosomes in eukaryotic cells. Increasing evidence suggests that EVs act as potent communicasomes, that is, nano-sized extracellular organelles that play diverse roles in intercellular communication. The biogenesis and functions of EVs may share many features in different biological systems. Thus, the study of EVs provides crucial keys to understanding the intercellular communication network in living organisms and the evolutionary connections between prokaryotes and eukaryotes. A wide variety of Gram-negative bacteria constitutively secrete EVs during growth. Bacterial EVs are composed of lipopolysaccharide, proteins, genetic materials, and other factors associated with virulence. Studies of EVs from diverse bacterial strains suggest their roles in the delivery of toxins to host cells, the transfer of proteins and genetic material between bacterial cells, cell-to-cell signals, and the elimination of competing organisms. Although growing evidence suggests that Gram-negative bacterial EVs are essential for bacterial survival and pathogenesis in hosts, the mechanisms of vesicle formation and of protein sorting into EVs, as well as the pathophysiological roles of EVs, have not been clearly defined. To address these issues, vesicular proteins should be comprehensively identified. Proteomics offers a powerful approach to decode the protein components of EVs. Using a proteomics approach, a comprehensive proteome map of Escherichia coli-derived native EVs has been established with high confidence. This information helps elucidate the biogenesis and functions of EVs from nonpathogenic and pathogenic bacteria. Although archaea, Gram-negative bacteria, and mammalian cells constitutively secrete EVs as a mechanism for cell-free intercellular communication, this cellular process has been overlooked in Gram-positive bacteria. In the present study, I found for the first time that Gram-positive bacteria naturally produce EVs into the extracellular milieu. This observation suggests that the secretion of EVs is an evolutionally conserved, universal process that occurs from simple organisms to complex multicellular organisms. With a proteomics approach, a total of 90 protein components of Gram-positive Staphylococcus aureus-derived EVs were identified with high confidence. In the group of identified proteins, I found key proteins that facilitate the transfer of antibiotic resistance proteins to other bacteria and pathological functions in systemic infectious diseases including coagulation and thrombosis disorders. In-depth study on the role of these EV-associated proteins revealed that S. aureus EVs play diverse roles in polymicrobial community as well as multiple roles for pathogenesis in an inter-species world. Studies of EVs will help us not only to elucidate the biogenesis and functions of EVs but also to stimulate the development of diagnostic tools, novel vaccines, and therapeutic agents which will advance both basic and clinical sciences.

쥐의 뇌조직으로부터 정제한 synaptic vesicles의 2-D PAGE와 MALDI-TOF/MS를 이용한 Proteome 연구

김혜정 연세대학교 대학원 2005 국내석사

Synaptic vesicle proteins and their markers are useful tools for the understanding of the complex life cycle of synaptic vesicles underlying intercellular communication. Whole proteomes of synaptic vesicle from rat brains were analyzed by using 2-dimentional electrophoresis (2-DE) and matrix-assisted laser desorptionionization-time of flightmass spectrometry (MALDI-TOF). In the present study, I have prepared a highly purified synaptic vesicle fraction from rat brains by sequentially differential centrifugation, hypoosmotic shock, and sucrose-density gradient centrifugation. Proteins were separated by isoelectric focusing on immobilized IPG DryStrips in the pH range of 3-10 (linear), and then by 10% gradient SDS-PAGE gels. Protein identification was done by peptide mass fingerprinting with MALDI-TOFMS. Yield of the synaptic vesicle fraction to brain homogenate was 0.18% and its purity was confirmed by using an electron microscope. This study identified the optimal voltage conditions of IEF for the separation of synaptic vesicle proteins. A total of 619 protein spots were obtained from 13 cm gels. Synaptic vesicle proteins were mostly located on low molecular weight range at pH ranges of 5-8. At present, all 64 proteins were identified from 2-DE and MALDI-TOFMS. The identified proteins involved in the biogenesis and trafficking of vesicle, energy metabolism, signal transduction, transport and unknown function. An updated map of rat brain proteins is presented and the method may be applied to a field for improving the limited proteomics approach to brain protein expression. 본 연구는 2-D PAGE와 MALDI-TOFMS를 이용하여 흰쥐의 뇌에서 whole synaptic vesicle에 대한 proteome map을 완성하고자 하였다. Differential centrifugation과 hypoosmotic shock, sucrose gradient centrifugation을 이용한 정밀한 정제과정을 거쳐 쥐의 뇌조직에서 고도로 정제된 synaptic vesicles을 분리하였으며, 분리된 synaptic vesicle의 양은 전체 단백질양의 0.18% (wv)에 해당하였다. 전자현미경으로 관찰한 결과 고도로 정제된 균질한 모양의 synaptic vesicles임을 확인 하였다. 정제된 synaptic vesicle 단백질 시료는 immobilized pH gradient strip (pH 3-10)을 사용하여 first-dimensional isoelectric focusing을 한 다음, 10% SDS-PAGE로 second-dimension electrophoresis를 실행하였다. Synaptic vesicle 단백질의 whole map을 얻기 위한 최적의 isoelectric focusing 조건을 확립 하였다. 얻어진 단백질은 in-gel digestion 후 MALDI-TOFMS를 거쳐 단백질을 확인하였다. 13 cm gel을 사용하여 619개의 단백질 spot을 얻었고 그 중에서 총 64개의 synaptic vesicle과 관련된 단백질을 확인하였다. 단백질 spot들은 대부분 50 kDa 이하의 비교적 낮은 분자량을 가진 것이었으며, pI값은 5-8의 범위에 걸쳐있었다. 확인된 단백질은 vesicle 운반과, signaling, 에너지 대사, 구조유지, 그 밖에 몇 종의 효소와 수용체 등의 기능으로 나눌 수 있었으며, 기능이 알려지지 않은 단백질도 다수 포함되어 있었다. 이 연구 는 synaptic vesicles을 분석하여 기존의 뇌조직에 존재하고 있는 단백질의 database를 향상시키고, 원심분리를 이용한 전처리 단계와 지질 성분이 풍부한 분획에 대한 최적의 isoelectric focusing 조건을 정립함으로써 proteomics 방법의 실험적 한계를 개선하는데 기여하였다고 사료된다. 또한 이 연구결과는 synaptic vesicle cycle을 포함하여 그들 사이의 정보 전달 기전을 이해하고 연구하는데 기여할 것으로 사료된다.

Investigation on the potential of antibiotic-resistant substances delivery via extracellular vesicles (EVs) from antibiotic resistance bacteria

이애린 경상대학교 대학원 2021 국내석사

Bacteria naturally release extracellular vesicles (EVs) during growth. These EVs are known as membrane vesicles (MVs) in Gram-positive bacteria or outer membrane vesicles (OMVs) in Gram-negative bacteria. EVs are known to store and to protect proteins, small molecules, and genetic materials and to deliver them. They play diverse roles, such as enabling bacterial survival during stress conditions and communications within bacterial communities. Here, this study shows that EVs from Gram-positive bacteria are possible to transmit antibiotic resistance substances unto antimicrobial-sensitive Gram-negative bacteria. We collected EVs from a methicillin-resistant Staphylococcus aureus (MRSA) called ST541. Using these EVs, we performed vesicles-mediated transformation of antimicrobial-sensitive Escherichia coli called RC85 based on established protocols. The transformant RC85 colonies that acquired antibiotic resistance were re-select on antibiotic selection plates. One of them was named RC85-T. The resulting transformant RC85 was exhibited logarithmic phase growth but slightly slower than RC85. RC85-T showed a higher value of resistance against β-lactam antibiotics when compared with minimum inhibitory concentrations (MICs) of RC85. OMVs from the RC85 and the RC85-T were purified, and their quantitative production rates and β-lactamase activity were compared. The production of both OMVs was nearly equal. The β-lactamase activity of OMVs from the RC85-T was higher than that of the RC85 OMVs per equal protein concentration. We performed a comparative proteomic analysis to investigate the difference in β - lactam resistive compounds carried by OMVs and investigated whether both types of OMVs could protect susceptible E.coli from β-lactam-induced death. Several proteins that can be involved in degrading β-lactam antibiotics were more abundant in OMVs from RC85-T. Also, OMVs from RC85-T dose-dependently rescued β-lactam-susceptible E. coli from β-lactam antibiotic-induced growth inhibition. In conclusion, we surmise that certain substances in Gram-positive ST541 EVs may be transferred to Gram-negative bacteria E. coli RC85, resulted in β-lactam antibiotic resistance. The findings of the present study might provide the importance of MVs in promoting the emergence of resistant strains out of a mixed community of bacteria.

Systematic Study on Microfluidic Formation of Polymeric Vesicles in Multiple Lamination Flow

응웬 부산대학교 대학원 2019 국내박사

Polymeric Micelles, especially Vesicles have made a significant contribution to the colloidal researches and their applications in many fields of drug delivery and biomedical study have attracted a lot of attention from research groups worldwide. This thesis presents systematically a study on microfluidic formation of the polymeric vesicles (PVs) in a multiple lamination flow generated in a double flow-focusing microchannel (DFFM). This formation process occurred in multiple steps from self-assembly of spherical micelles on the flow interface to the fusion and growth of spherical micelles to form the PVs. The meeting of the polymer stream and water stream through the transverse diffusion of water and polymer molecules activated the self-assembly to form spherical micelles in the beginning. This was simulated and visualized by using a 3D bond fluctuation method. The growth process of spherical micelles to form PVs were also investigated thoroughly and confirmed by SEM images after the microchannel was frozen with liquid nitrogen. Along with the formation of PVs, the DFFM can overcome the current issue of channel clogging in common microfluidic synthesis using single flow-focusing microchannel (SFFM) and produces PVs with high uniformity in size. In addition, the influenced factors of the formation process of PVs were studied. Finally, the produced PVs were tested for potential OLED and encapsulation applications.

뱀 골격근 근소포체 칼슘 유리 채널의 특성에 관한 연구

남장현 충남대학교 1996 국내박사

To investigate properties of Ca-release channel in the reptile skeletal muscle, electrophoretical analysis, purification of RyR, [³H]ryanodine binding study, and ^45Ca-release were carried out in the SR vesicles which were prepared from the snake skeletal muscle. Results are as follows ; 1.The snake SR vesicle has the single high molecular weight protein band on SDS polyacrylamide gel(PAGE), and its mobility was similar with that of rat skeletal SR vesicles. 2.The high molecular weight band on SDS PAGE was found in the [³H]ryanodine peak fractions (Fr_5-7) obtained from the purification step of the RyR. 3.Maximal binding site and K_D of the snake SR RyR were 6.36 pmol/mg protein and 17.62 nM, respectively. Specific binding of [³H]ryanodine was significantly increased by calcium and AMP (P < 0.05), but not or slightly inhibited by tetracaine, ruthenium red (5.4%), or MgC1₂(21%). 4.^45Ca-release from the SR vesicles loaded passively was significantly increased by the low concentration of calcium (1 - 10 μM) and AMP(5 mM)(P <0.05), but significantly decreased by the high concentration (300 μM) of calcium, tetracaine(1 mM), ruthenium red (10 μM), and MgC1₂(2 mM)(P<0.05). 5.Ryanodine receptor of the snake skeletal muscle is reacted with polyclonal (rabbit serum) to rat RyR, but not with monoclonal Ab(mouse ascites) to rat RyR. From the above results, it is suggested that snake SR vesicles also has the RyR showing the similar properties to those of mammalian skeletal RyR with the exceptions of lower values of [³H]ryanodine maximal binding sites and K_D and of no or slight inhibition of [³H]ryanodine-binding by tetracaine, ruthenium red, or MgCl₂.

Attenuated but immunologically potentiated tumor extracellular vesicles as cancer vaccines

한지훈 KU-KIST Graduate School of Converging Science and 2022 국내석사

Tumor extracellular vesicles (TEVs) are lipid bilayer particles that are secreted by tumor cells, and are known to play a key role in tumor cell to cell communications, mostly promoting tumor progression. Since TEVs reflect the characteristics of its original tumor cells, they have attracted much attention for their applications in biomarkers, drug delivery vehicles and notably, vaccines. Cancer vaccines are a mode of cancer immunotherapy that is designed to specifically activate the immune system to combat cancer in either a prophylactic or a therapeutic way. However, according to numerous studies signing TEVs as a tumor-promoting communicator between cancer cells, TEVs seem to be a double-edged sword when it comes to their utility as cancer vaccines. In this study, we have used the drug verteporfin in order to attenuate the tumorigenic characteristics yet potentiate the immunogenic properties of TEVs, thereby designing an effective cancer vaccine. This strategy lead to an attenuated but immunologically potentiated TEVs (AI-TEVs) that were silenced of the tumorigenic properties of TEVs yet had heightened antigenic and adjuvantic qualities, to be used as prophylactic cancer vaccines. Tumor extracellular vesicles (TEVs) 은 암세포에 의해 분비되는 지질이중층 입자들이며, 주로 암 성장을 촉진시키는 방향으로 암세포들 간의 소통을 주관하는 역할을 하는 것으로 알려져 있다. TEV들은 유래한 암세포의 특징을 반영하기 때문에 바이오마커, 약물 수송 수단 그리고 특히 암 백신으로의 활용에 대한 주목을 많이 받고 있다. 암 백신이란 암 면역 치료의 수단 중 면역체계를 특이적으로 활성화 시켜 암을 예방하거나, 혹은 치료할 수 있도록 한다. 그러나 TEV가 암을 촉진시킨다는 다수의 연구들을 참고하면 TEV는 암 백신으로 응용함에 있어서 양날의 검으로 보인다. 이 연구에서는 Verteporfin이라는 약물을 사용하여 TEV의 암 촉진적인 특징들은 억제하고, 동시에 면역학적인 특징들은 증진시켜 효과적인 암 백신으로 만들고자 하였다. 이 전략에 의해, 일반적인 TEV들의 암 촉진적인 성질은 잠재워졌으며 항원성과 애주번트성은 증가되어, 예방적인 암 백신으로서 효과적으로 사용될 수 있는 attenuated but immunologically potentiated TEVs (AI-TEVs)를 개발할 수 있었다.