정상 공여자에서 지속적인 G-CSF 정맥내 투여에 의한 CD34+ 세포의 가동화

이 석,성주명 大韓輸血學會 1999 大韓輸血學會誌 Vol.10 No.2

골수유핵세포의 경내피세포 및 경기질세포 이행 과정에서 류코트리엔 B_(4) 역할과 활성산소종 억제의 영향

문영철,오수아,유은선,최문영,안지영,성주명 이화여자대학교 의과학연구소 2008 EMJ (Ewha medical journal) Vol.31 No.2

Objectives:Leukotriene B_(4)(LTB4) is lipid mediator derived from membrane phospholipids during the process of inflammation, having many roles(ie;inducer of chemotaxis, the production of nitric oxide, transepithelial migration of neutrophil). The major activities of LTB4 include the recruitment and activation of leukocytes, suggesting that it may involve the process for transendothelial migration of nuclear cells in bone marrow environment. Reactive Oxygen Species (ROS) have a cell signaling roles that are involved in signal transduction cascades of numerous growth factor-, cytokine-, and hormone-mediated pathways, and regulate many biological systems. In this present study, we focused on the role of LTB4 and ROS on transmigration of bone marrow nuclear cells across endothelial or stromal cell monolayer. Methods:MS-5, murine stromal cell line cells, or bEnd.3, murine microvascular cell line cells, were grown to confluence on microporous transwell membrane. Murine marrow cells were placed on top of the prepared transwell membrane. The transwells were then seated in wells containing media and LTB4 with or without pretreatment of N-acetylcysteine(NAC), an oxygen free radical scavenger, or diphenylene iodonium(DPI), an inhibitor of NADPH oxidase-like flavoproteins. Cells that migrated through the stromal or endothelial layer into the wells were assayed for transendothelial migration. Results:The numbers of migrated bone marrow nuclear cells through the bEnd.3 were increased by treatment of LTB4(control, 12.5±0.2%;50nM, 22.7±0.9%;100nM, 44.3±1.4%;200nM, 36.3±0.9%;p<0.05). The numbers of migrated bone marrow nuclear cells through the MS-5 were also increased by treatment of LTB4(control, 11.0±0.9%;50nM, 25.7±0.9%;100nM, 35.8±1.8%;200nM, 32.1±0.9%;p<0.05). However, increasing effect of LTB4 to the transmi-gration of bone marrow nuclear cells through the MS-5 or bEnd.3 were inhibited by pretreatment of NAC or DPI. Conclusion:Through our data, it is suggested that LTB4 could induce the transmigration of bone marrow nuclear cells and ROS might be involved on the transendothelial migration of bone marrow nuclear cells by LTB4. It would be very interesting to test the effects of LTB4 and ROS on stem cell mobilization and homing in the future. 류코트리엔 B_(4)(LTB4)는 세포막 인지질로부터 유래된 염증과정에 관여하는 지방매개체이다1-3). LTB4의 주된 역할은 화학주성의 유도, 활성산소의 생산, 백혈구, 특히, 호중구 및 호산구의 이동에 관여하고 있으며, 내피세포와 백혈구의 부착에 중요한 역할을 하는데, 이러한 LTB4의 역할은 케모카인에 의한 백혈구의 이동과 화학주성과 많은 부분에서 유사한 특성을 가진다4-7). 염증반응과 백혈구 이동과 연관된 LTB4의 역할들은 골수내 세포들의 이동 과정에서 다른 시토카인이나 성장인자 혹은 케모카인의 역할과 유사한 점이 많다. LTB4와 비슷한 작용을 가진 여러 케모카인들이 조혈모세포를 포함한 골수내 세포들의 이동과 연관이 있음이 최근 밝혀짐에 따라, LTB4도 골수내 세포의 이동에 관련이 있을 가능성이 있다8-14). 활성산소종(ROS)은 여러 종류의 성장인자, 시토카인, 혹은 호르몬 등으로 유도된 조혈세포내 신호전달체계에 중요한 역할을 하고, 조혈세포내 여러 생명현상의 조절과 관련이 있다15-18). 특히, 염증세포에서 분비된 시토카인 혹은 케모카인들이 세포내 ROS를 증가시키고, 이때 증가된 ROS는 혈관내피세포 단층의 투과성을 증가시켜 백혈구의 경내피세포 이행을 용이하게 한다19-22). 조혈모세포 이식 후 조혈모세포가 말초혈액에서 골수 내로 자리잡는 귀소나, 골수내 조혈모세포를 혈액으로 나오게 하는 가동화의 과정은 매우 복잡하여, 여러 다양한 기전들이 관련이 있지만, 혈관내피세포 및 기질세포와 골수 조혈세포들과의 유기적인 반응과 골수 조혈세포의 경내피세포 이행이 중요한 역할을 한다23)24). 본 연구에서는 골수유핵세포의 경내피세포 및 경기질세포 이행에 LTB4와 ROS가 어떻게 영향을 미치는지 알아보고자 하였다.

침습성 아스페르길루스증으로 치료받았던 급성 백혈병 환자에서 조혈모세포이식후 발생한 파급성 아스페르길루스증 1례

배기선,박지영,신수연,문영철,최희정,조민선,성주명 대한감염학회 2003 감염과 화학요법 Vol.35 No.4

본 증례는 침습성 아스페르길루스증의 과거력을 갖고 있는 백혈병환자에서 조혈모세포이식을 받은 후 치명적인 파급성 아스페르길루스증이 발병한 예이다. 환자는 항암요법 후 흉부 방사선 및 단층촬영에서 진균성 폐렴을 의심하여 항진균제를 투여하고 폐엽절제술을 시행하여 아스페르길루스에 의한 폐렴임을 확인하였다. Amphotericin B로 치료한 후 조혈모세포이식을 시행받은 뒤 치료에도 불구하고 파급성 아스페르길루스증으로 진행되어 사망하였다. 이 증례에서 보듯이 이전에 아스페르길루스증의 과거력이 있는 경우에 이식후 치명적인 결과를 유발할 수 있으므로, 고위험군을 선별하는 지침과 적절한 치료법에 대한 연구가 필요하겠다. Invasive aspergillosis has been increasing as the number of severe immunocompromised hosts rises. Particularly, in allogeneic hematopoietic stem cell transplantation (HSCT) recipients, incidence of invasive aspergillosis ranges from 4 to 10%. Even with appropriate treatment, the prognosis of invasive aspergillosis in allogeneic HSCT recipients remains poor, showing high mortality rate. Herein. we report a case where invasive aspergillosis in a patient with acute myelogeneous leukemia progressed to disseminated aspergillosis after allogeneic HSCT. A 31-year-old woman with acute myelogenous leukemia had invasive aspergillosis after third reinduction chemotherapy. After administering amphotericin B, the patient underwent the wedge resection of lung. and HLA-matched allogeneic HSCT was then conducted. On day 14 of transplantation, the patient died of disseminated aspergillosis, including possible cerebritis and endocarditis despite the amphotericin B therapy.

Clinical Applications of Stem Cells derived from Mobilized Peripheral Blood and Human Cord Blood

Seong, Chu Myong 이화여자대학교 세포신호전달연구센터 2005 고사리 세포신호전달 심포지움 Vol. No.7

Hematopoietic stem cells(HSC) have been used for curative purposes on leukemia, bone marrow aplasia(ie, aplastic anemia) and metabolic disorder. Though bone marrow has been a rich source for hematopoiesis, mobilized peripheral blood by cytokines proved efficient in terms of long term engraftment after myeloablative therapy. In fact, peripheral blood stem cell is superior shortening the period of cytopenia after myeloablative conditioning regimens(ie, Busulfan, Total Body Irradiation(TBI)) compared to using bone marrow for transplantation. While mobilized hematopoietic stem cells are used often, the mechanisms on mobilization from bone marrow to peripheral blood are not elucidated yet. So far, important players in these processes are; 1) VLA-4/VCAM-1, LFA-1/ICAM-1, 2) SDF-1/CXCR4, 3) protease(ie, neutrophil elastase, cathepsin-G, MMP-9. The limitation of using G-CSF, which is the most popular cytokine in the clinic for HSC mobilization is as follows; 1) side effects(muscle pain, weakness and occasional leukostasis), 2) slowness(it takes 4-6 days to have optimal mobilization). In result, it is necessary to find new molecules for the shortening of optimization(ideally few hours) and molecules with minimal side effects, but at the same time providing large amount of HSC. VE-cadherin has a role in vascular integrity among vascular endothelial cells. In our Lab, we found the decreased expression of VE-cadherin during G-CSF mobilization. When G-CSF was given with anti-VE cadherin, the speed of mobilization was faster compared to that of G-CSF alone. In vitro, the expression of BLT2 was upregulated on nucleated cells in the bone marrow when G-CSF was given. In addition, we were able to observe maximal mobilization within 4hours after an injection of LTB4 in the murine model. In 1989, Gluckman et al found that human cord blood(HCB) was a rich source of HSC and HCB was used for an allo-graft after a conditioning regimen for a Fanconi anemia patient. More than 15 years later, this patient is still in complete remission maintaining allo-graft. In addition, HCB has many attractions; 1) readily available, 2) less exposed to infective organisms, 3) more primitive nature, 4) less GVHD(Graft Versus Host Disease) after allogeneic transplantation. But HCB has a critical limitation, which is a low number of HSC in HCB for transplantation in adult patients. In the midst of developing an effective ex-vivo expansion system of HCB, we found that Hyaluronic acid(HA), ligand of CD44, had a ability for expanding more cells but at the same time, we found more apoptosis during this process. The methodes for maintaining sternness of HSC but minimizing apoptosis is being investigated during ex-vivo expansion system. Meanwhile, we are also trying to develop better ways for inducing early and late endothelial progenitor cells which can be used for angiogenesis and myogenesis using different cytokines and from different graft sources including HCB. By understanding the mechanisms of the mobilization of HSC in depth, we will have a large amount of HSC and at the same time a speedy way to mobilize HSC. Doing that, we will be in a good position in graft engineering for cell therapies. In addition, via an effective and smart exvivo expansion system on HCB, we are able to provide more relevant cells for the purposes on tissue repair.

Modulation of ex-vivo expanded human cord blood graft and mobilization of peripheral blood progenitor cells

Seong, Chu-Myong 이화여자대학교 세포신호전달연구센터 2003 고사리 세포신호전달 심포지움 Vol. No.5

Human umbilical cord blood(CB) is an alternative source of hematopoietic stem cell(HSC). However, the use of CB is limited by a delayed engraftment due to insufficient number of HSCs. Cytokine-mediated ex vivo expansion has been proposed as a means of increasing the number of CB HSCs for transplantation. However, the biology of CB HSCs during cytokine-mediated ex vivo expansion, such as apoptosis or expression of adhesion molecules, has not yet been elucidated. Human cord blood ex-vivo exansion system has been established using TPO, FL and G-CSF with/without feeder layer. In addition, by understanding the molecular mechanisms of apoptosis during ex-vivo expansion, we are able to build up more efficient ex-vivo system. In result, we can provide not only more hematopoietic stem cells(quantity) but "true stem cells" having better qualities. Bone marrow hematopoietic stem cell transplantation has been very powerful curative method for disease, i.e., leukemia, aplastic anemia and metabolic disease. Meanwhile, peripheral blood hematopoietic stem cells have been alternative sources for allogeneic transplantation. However, it takes 5-6 days to get optimal mobilization using cytokines like G-CSF. Though many transplantation centers have been using mobilized peripheral blood hematopoietic stem cell for allograft on curing hematopoietic malignancies, the optimal protocols for mobilization and its mechanisms on mobilization need to be elucidated. We would like to focus two areas. First, we would like to set up the speedy methods for mobilization using different cytokines, anti CAMs and proteinase inhibitors. Same time, we would like to figure out the way how to collect the large number("Mega" dose) of CD34+ cells. Second, the molecular mechanism of mobilization and its signal transduction pathways will be investigated. In results, we may apply our knowledges that we will obtain via this current projects to the clinic for the situations that we need to overcome the MHC barries(HLA-mismatch setting) with megadose CD34+ cells. And understanding the molecular mechanisms on mobilization may provide the insight for finding more specific molecules with having less sides effects on mobilization in near future.

제49차 대한내과학회 추계학술대회초록집 : 심포지움 ; 급성 백혈병 : 급성백혈병 치료의 발전 - 동종 및 자가 말초 조혈모세포 이식 -

성주명 ( Chu Myong Seong ) 대한내과학회 1997 대한내과학회지 Vol.53 No.-

제49차 대한내과학회 추계학술대회초록집 : 심포지움 ; 급성 백혈병 : 급성백혈병 치료의 발전 - 동종 및 자가 말초 조혈모세포 이식

성주명 ( Chu Myong Seong ) 대한내과학회 1997 대한내과학회지 Vol.53 No.2S

만성 저산소증이 가로 심혈관계에 미치는 영향

주영명,지제근,이계열 공군본부 의무감실 1968 항공의학 Vol.16 No.1-2

How Light-Absorbing Properties of Organic Aerosol Modify the Asian Summer Monsoon Rainfall?

Chu, Jung-Eun,Kim, Kyu-Myong,Lau, William K. M.,Ha, Kyung-Ja American Geophysical Union 2018 Journal of Geophysical Research: Atmospheres Vol.123 No.4

난소암 세포주에서 Topotecan, Cisplatin, Taxol에 대한 화학민감도의 측정과 p53, bcl-2 및 세포고사에 관한 연구

허주엽 ( Huh Chu Yeop ),임명철 ( Lim Myong Cheol ),서병선 ( Suh Byung Sun ) 대한산부인과학회 2003 Obstetrics & Gynecology Science Vol.46 No.7

목적 : 본 연구는 난소암 세포주에서의 Topotecan, Cisplatin, Taxol에 의한 세포고사를 통하여 화학민감도를 측정하고 세포고사가 일어날 때 p53유전자와 bcl-2유전자와의 상호관계를 평가하기 위함이다. 연구 방법 : 이번 연구에서 저자들은 Topotecan, Cisplatin, Taxol을 5가지 인간 난소암 세포주에 반응시켜서 유도된 세포고사의 결과를 얻었고, Topotecan, Cisplatin, Taxol의 여러 농도에서의 단독, 복합 또는 연속적 투여에 대한 민감도를 조사하였다. 또한 난소암 세포주에 대한 이들 화학요법의 상호작용과 세포 사망의 정도를 정량적으로 분석을 통한 세포고사와 세포독성을 비교하였다. 세포고사가 일어날 때 p53유전자와 bcl-2유전자가 변하는가를 알기 위해 western blotting으로 유전자 검사를 시행하였다. 결과 : 5가지 세포주는 Topotecan, Cisplatin, Taxol (LD_50 range of Topotecan, 30~1000 ng/ml; Cisplatin, 3~10 ㎍/ml; Taxol 5~1000 nm)에 대한 다양한 민감도를 보여주고 있다. SKOV-3는 다른 세포주에 비하여 저항성이 있는 세포주로서 Topotecan에 대한 저항성은 다른 세포주에 비해 2~30배, Cisplatin에서는 3배, Taxol에서는 200배 저항성을 보였다. Topotecan, Cisplatin, Taxol의 세포주에 대한 민감도와 세포고사와의 관계가 SNU-840과 OVCAR-3에서 관찰되었다. Topotecan, Cisplatin, Taxol 24시간 혹은 48시간 처리시 OVCAR-3의 DNA 분절은 동일하게 나타났다. 세포독성 분석과 연관지어 염기순서분석 실험을 시행하였을 때, SNU-251을 제외한 다른 세포주에서 Topotecan, Cisplatin, Taxol간의 다른 상호관계에서 의미있는 차이를 보이지 못하였다. topotecan을 전처치하였을 경우, Cisplatin을 투여한 군에서 24시간째 세포독성이 증가하였다. 전기영동검사상 fragmented DNA의 정량화를 통하여 동일한 결과를 보인다. 결론 : Topotecan, Cisplatin, Taxol에 의해서 의미있는 세포독성이 생기는 것은 단순한 괴사에 의한다기 보다는 직접적인 세포고사의 유도와 관련이 있음을 확인하였고, 이들 약제에 대한 치료성적은 세포적 특성, 유전적 특성의 차이에 의함을 알 수가 있었다. Objective : The aim of this study was to evaluate Topotecan-, Cisplatin- and Taxol-induced apoptosis in five human ovarian cell lines as a measure of chemosensitivity and relationship between apoptosis and p53 and bcl-2 gene expression. Methods : In this study, the author is presenting data on apoptosis induced by Topotecan, Cisplatin and Taxol in five ovarian cancer cell lines, and represent different levels of sensitivities to Topotecan, Cisplatin and Taxol. This study also includes the interaction of these chemotherapeutic agents on ovarian cancer cell lines with respect to the apoptosis and cytotoxicity assay as a quantitative measure of the efficiency of killing. Presence of the p53 and bcl-2 gene product were examined by western blotting. Results : The five cell lines represent various sensitivities to Topotecan, Cisplatin, Taxol (LD_50 range of Topotecan, 30~1000 ng/ml; Cisplatin, 3~10 ㎍/ml, Taxol 5~1000 nm). SKOV-3 represent a resistant cell line which was 2~30 times resistant to Topotecan, 3 times resistant to Cisplatin, and 2~200 times resistant to Taxol when compared to others. Demonstration of apoptosis correlated with the sensitivity of the cell lines to Topotecan, Cisplatin and Taxol for SNU-840 and OVCAR-3. DNA fragmentation of OVCAR-3 was uniformly present when treated with Topotecan, Cisplatin and Taxol, 24 or 48 hours. When sequencing experiments were performed with correlated with cytotoxicity assays, except in SNU-251 cells where no significant difference was observed in different interactions of Topotecan, Cisplatin and Taxol. Pretreatment with Topotecan, Cisplatin at a 24 hour interval resulted in enhanced cytotoxicity. Quantitation of the fragmented DNA correlated with that seen on gel electrophoresis. Conclusion : The study indicate that the ability to achieve significant cytotoxicity by Topotecan, Cisplatin and Taxol may be related to the induction of apoptosis rather than necrosis. However, outcome of these treatments depend on cellular and genetic characheristics.