Advanced T and Natural Killer Cell Therapy for Glioblastoma

Wan-Soo Yoon,정동섭 대한신경외과학회 2023 Journal of Korean neurosurgical society Vol.66 No.4

Although immunotherapy has been broadly successful in the treatment of hematologic malignancies and a subset of solid tumors, its clinical outcomes for glioblastoma are still inadequate. The results could be due to neuroanatomical structures such as the bloodbrain- barrier, antigenic heterogeneity, and the highly immunosuppressive microenvironment of glioblastomas. The antitumor efficacy of endogenously activated effector cells induced by peptide or dendritic cell vaccines in particular has been insufficient to control tumors. Effector cells, such as T cells and natural killer (NK) cells can be expanded rapidly ex vivo and transferred to patients. The identification of neoantigens derived from tumor-specific mutations is expanding the list of tumor-specific antigens for glioblastoma. Moreover, recent advances in gene-editing technologies enable the effector cells to not only have multiple biological functionalities, such as cytokine production, multiple antigen recognition, and increased cell trafficking, but also relieve the immunosuppressive nature of the glioblastoma microenvironment by blocking immune inhibitory molecules, which together improve their cytotoxicity, persistence, and safety. Allogeneic chimeric antigen receptor (CAR) T cells edited to reduce graft-versushost disease and allorejection, or induced pluripotent stem cell-derived NK cells expressing CARs that use NK-specific signaling domain can be a good candidate for off-the-shelf products of glioblastoma immunotherapy. We here discuss current progress and future directions for T cell and NK cell therapy in glioblastoma.

Diethylamino-curcumin mimic with trizolyl benzene enhances TRAIL-mediated cell death on human glioblastoma cells

안용철,Seokjoon Lee,Cheon Soo Park,Hyuk-Jai Jang,이지환,Byong-Gon Park,Yoon Sun Park,Woon-Seob Shin,Daeho Kwon 대한독성 유전단백체 학회 2018 Molecular & cellular toxicology Vol.14 No.2

Backgrounds: Glioblastoma multiforme is one of the most aggressive human malignant brain tumors. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is known as the death ligand, which induces preferential apoptosis of transformed cancer cells. In this study, we demonstrated that the newly synthesized diethylamino-curcumin mimic with trizolyl benzene (YM-4) enhances cytotoxicity in combination with TRAIL in human glioblastoma cells. Methods: We synthesized diethylamino-curcumin mimic with trizolyl benzene (YM-4) and investigated possible apoptotic cell signaling by co-treatment with YM-4 and TRAIL on human glioblastoma cells. Results: Caspase-8, 9, and 3 and poly (ADP-ribose) polymerase were more efficiently cleaved with cotreatment of YM-4 and TRAIL than treatment with each alone in human glioblastoma cells. Co-treatment with YM-4 and TRAIL significantly increased the expression of Bax and Smac/Diablo and also inhibited the expression of the X-linked inhibitor of apoptosis protein and Survivin in human glioblastoma cells. Conclusion: These results demonstrated that YM-4 can be an anticancer candidate that can be effective on human glioblastoma cells in combination with TRAIL.

Rapid and Quantitative Measurement of Cell Adhesion and Migration Activity by Time-series Analysis on Biomimetic Topography

심우광,차정화,최철희,최경선 한국생물공학회 2017 Biotechnology and Bioprocess Engineering Vol.22 No.2

Glioblastoma Multiforme (GBM) is the most malignant brain tumor in adults, highly infiltrative and difficult to cure. According to the histopathological evidence, the glioma cells are found to infiltrate into the surround normal brain tissue, along the Scherer’s structure (e.g. white matter tract and microvasculature). As a major invasion route of microenvironments, these pre-existing anatomic structures should be considered in studying infiltrative movement of glioblastoma. In our previous work, we introduced in vitro biomimetic platform as alternative model of brain-anatomical structures to study about migratory phenotypes of glioblastoma. By applying this proper biomimetic platform, we further investigated the influence of integrin, which is one of mechanoreceptors to sense mechanical cues, on phenotype of glioblastoma cells in this study. On in vitro biomimetic platform, glioblastoma cells show elongated morphology with highly aligned along the patterned direction, which is similar to that on in vivo condition. These morphological changes were gradually progressed in time-dependent manner, which might be mediated by a representative mechanoreceptor, integrin. Treatment of cell adhesive motif for integrin inhibition hinders the morphological dynamics on in vitro biomimetic platform in early time-point compared with cell proliferation cycle. Since cell adhesion mediated by mechanoreceptors is one of essential steps in migration/invasion, our results imply that effect of integrin on glioblastoma invasion is mediated by the mechanosensing process on topography and indicated by morphological changes. For further application, this quantitative analysis of glioblastoma morphology on biomimetic platform can be contributed to simple and ease investigation and effective anti-cancer drug screening.

교아세포종에 대한 면역치료 전략

윤미정 ( Mi Jung Yun ),한상배 ( Sang Bae Han ),남도현 ( Do Hyun Nam ),김충현 ( Choong Hyun Kim ),김정훈 ( Jeong Hoon Kim ),최종성 ( Jong Sung Choi ),김환묵 ( Hwan Mook Kim ),임재승 ( Jae Seung Lim ) 대한뇌종양학회·대한신경종양학회·대한소아뇌종양학회 2007 대한뇌종양학회지 Vol.6 No.2

Despite efforts and advances in surgical resection, radiation, and chemotherapy, the mean survival time of glioblastoma remained little changed during the last few decades. To overcome this situation, immune cell therapy has been attempted in various animal models and human cancer patients. Immunotherapy for glioblastoma aims at enhancing immunogenicity of glioma cells and cytotoxic activity of immune effector cells. The present review focuses on immune-mediated treatment approaches for glioblastoma and therapeutic trials. Especially, we address the usefulness of cytokine-induced killer cells(CIK cells) for treatment of glioblastoma patients.

Kaposi’s Sarcoma-Associated Herpesvirus Infection Modulates the Proliferation of Glioma Stem-Like Cells

( Hyungtaek Jeon ),( Yun Hee Kang ),( Seung-min Yoo ),( Myung-jin Park ),( Jong Bae Park ),( Seung-hoon Lee ),( Myung-shin Lee ) 한국미생물생명공학회(구 한국산업미생물학회) 2018 Journal of microbiology and biotechnology Vol.28 No.1

Glioblastoma multiforme is the most lethal malignant brain tumor. Despite many intensive studies, the prognosis of glioblastoma multiforme is currently very poor, with a median overall survival duration of 14 months and 2-year survival rates of less than 10%. Although viral infections have been emphasized as potential cofactors, their influences on pathways that support glioblastoma progression are not known. Some previous studies indicated that human Kaposi’s sarcoma-associated herpesvirus (KSHV) was detected in healthy brains, and its microRNA was also detected in glioblastoma patients’ plasma. However, a direct link between KSHV infection and glioblastoma is currently not known. In this study, we infected glioblastoma cells and glioma stem-like cells (GSCs) with KSHV to establish an in vitro cell model for KSHV-infected glioblastoma cells and glioma stem-like cells in order to identify virologic outcomes that overlap with markers of aggressive disease. Latently KSHV-infected glioblastoma cells and GSCs were successfully established. Additionally, using these cell models, we found that KSHV infection modulates the proliferation of glioma stem-like cells.

좌측 전두엽에 발생한 거대 세포 교모세포종 -증례보고-

홍용표 ( Yong Pyo Hong ),김문규 ( Moon Kyu Kim ),오세문 ( Sae Moon Oh ),박세혁 ( Se Hyuck Park ) 대한뇌종양학회·대한신경종양학회·대한소아뇌종양학회 2006 대한뇌종양학회지 Vol.5 No.2

Giant cell glioblastomas are defined as glioblastomas with marked predominance of bizarre and mutinucleated giant cells. They represent about 5% of all glioblastomas and can occur at any site of the central nervous system. Giant cell glioblastomas show a prolonged survival period compared with usual histological type of glioblastomas. The authors report a 23-year-old patient who had undergone resection and adjuvant radio-chemotherapy for a giant cell glioblastoma on left frontal lobe. The patient survived for 3.9 years.

Force-mediated proinvasive matrix remodeling driven by tumor-associated mesenchymal stem-like cells in glioblastoma

( Eun-jung Lim ),( Yongjoon Suh ),( Seungmo Kim ),( Seok-gu Kang ),( Su-jae Lee ) 생화학분자생물학회 2018 BMB Reports Vol.51 No.4

In carcinoma, cancer-associated fibroblasts participate in force-mediated extracellular matrix (ECM) remodeling, consequently leading to invasion of cancer cells. Likewise, the ECM remodeling actively occurs in glioblastoma (GBM) and the consequent microenvironmental stiffness is strongly linked to migration behavior of GBM cells. However, in GBM the stromal cells responsible for force-mediated ECM remodeling remain unidentified. We show that tumor-associated mesenchymal stem-like cells (tMSLCs) provide a proinvasive matrix condition in GBM by force-mediated ECM remodeling. Importantly, CCL2-mediated Janus kinase 1 (JAK1) activation increased phosphorylation of myosin light chain 2 in tMSLCs and led to collagen assembly and actomyosin contractility. Collectively, our findings implicate tMSLCs as stromal cells providing force-mediated proinvasive ECM remodeling in the GBM microenvironment, and reminiscent of fibroblasts in carcinoma. [BMB Reports 2018; 51(4): 182-187]

Effect of glioblastoma-cultured medium on neural differentiation of porcine induced pluripotent stem cells

Eunhye Kim,Young Seok Park,Sang-Hwan Hyun 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

Little is known to date about neural development of pig and directed differentiation of porcine pluripotent stem cells (PSCs) to neuronal cells remains elusive. To determine whether soluble factors from glioblastoma multiforme (GBM) promoted the neural differentiation from porcine induced PSCs (iPSCs), cells were treated cultured media of GBM cells. First of all, we isolated and established primary GBM cell line (WHO grade IV). The cellular morphology of GBM cancer cell line are dendritic-like with positive expression in NESTIN, SOX2, VIMENTIN and GFAP using immunofluorescence analysis. G-banded karyotype from primary GBM cell line revealed severe numerical chromosomal aberrations. GBM-cultured medium (CM) treated iPSC-NPCs survive well in vitro when supplemented with a combination of growth factors, including EGF and bFGF. The GBM-CM treated differentiated cells showed an increased mRNA expression level of astrocyte marker, GFAP and the dopaminergic neuron marker, tyrosine hydroxylase (TH). However, there was no significant difference in mRNA expression level of oligodendrocyte marker, MBP. The protocol developed in the present study for large animal models might provide an exciting tool to bridge the present gaps in neuroscience studies between rodents and humans.

수아세포종 세포와 교모세포종 세포의 배양에서 all-trans와 13-cis retinoic acid의 세포증식억제 효과

윤수한(Soo Han Yoon),김세혁(Se Hyuk Kim),안영환(Young Hwan Ahn),안영민(Young Min Ahn),조기홍(Ki Hong Cho),조경기(Kyung Gi Cho),김성환(Sung Hwan Kim) 대한소아신경학회 1998 대한소아신경학회지 Vol.5 No.2

목적 : 암화학요법중에 대표적인 약물중의 하나인 레티노익 산을 이용하여 신경모세포종과 교모세포종을 포함하여 여러 암세포에서 증식억제를 이룰 수 있다고 하지만, 최근에 악성 교종 세포종에서의 임상적 시도에서 약물반응도가 낮았다는 상반된 결과를 해결하기 위해 혈중 유효약물농도에서 종양증식억제 정도를 측정해보고자 하였다. 또한, 레티노익 산의 작용이 종양세포의 분화를 촉진시키고 종양의 증식을 억제하는 것이라면 당연히 성인종양보다는 소아종양에서 효과가 기대되는 만큼 뇌종양에서는 무엇 보다도 대표적인 소아종양인 수아세포종에서 효과를 확인해보고자 하였다. 대상및 방법 : 수아세포종 환자의 생검조직에서 배양된 수아세포종과 교모세포종 373과 87세 포주를 배양하여 all-trans 와 13-cis 레티노익 산의 농도를 10¯5M에서 10¯¹¹M까지 10¯5M, 10¯6, 10¯7, 10¯8, 10¯9, 10¯10 및 10¯¹¹M등의 7단계로 나누고 4일, 7일 그리고 14일간 배양후 각 단계별 세포성장억제(cell growth inhibition) 정도를 MTT 검사에 의해 비교하였다. 결과 : 87세포주에서 13-cis 레티노익 산의 경우에 10¯8M부터 10¯7M까지에서는 10~15%의 종양증식억제가 이루어졌지만 10¯6M에서 10¯5M의 농도에서는 증식억제가 4일에는 10-22%, 7일에는 10%, 14일에는 0~12%였고, all trans 레티노익산의 경우에는 10¯6M 이하 농도에서는 4일, 7일, 그리고 14일 모두에서 5% 미만의 증식억제를 보였는데 10¯5M 농도에서는 4일과 7일에는 증식억제가 42%와 37%였지만, 14일에는 0%로서 시간 경과에 따라 증식억제가 감소하였다. 373 세포주에서는 10¯6M과 10¯5M 농도의 all-trans 레티노익 산에 의해서 10¯6M과 10¯5M 농도에서 4일과 7일에는 5% 이하의 증식억제를 보이다가 14일에는 10-15%의 증식억제를 보였으며 13-cis 레티노익 산의 경우에는 10¯6M과 10¯5M에서 4일에는 약 30%, 7일에는 약 20%, 그리고 14일에도 약 20%의 증식억제를 보였다. 수아세포종에서는 10¯6M 이하 농도의 all-trans와 13-cis 레티노익 산에 의해서 증식억제가 약 25% 이상 이루어졌으며, 10¯6M 이상 농도에서 13-cis 레티노익 산의 경우 25% 정도의 증식억제를 보였지만 all-trans 레티노익 산에서는 약 40%의 증식억제를 볼 수 있었다. 결론 : 교모세포종의 경우 인체에 투여가능한 정도의 all-trans 또는 13-cis 레티노익 산 농도에서 각 세포주의 반응도가 달라서 그 효과를 예측할 수 없지만, 수아세포종에서는 13-cis 와 all-trans 레티노익 산 모두에서 종양 증식억제 효과가 있었으며, 특히 all-trans 이형체가 효과가 더 좋았으며, 또한 지속적 투여와 저농도(10¯9M 이하)에서도 종양증식 억제에 효과가 있었다. propose : It has been reported that retinoic acid, one of the most popular agents for chemoprevention could inhibit the proliferation of many cancer cells including neuroblastoma and glioblastoma. However, there is increasing demand reaccessing its in vitro inhibitory errect on the tumor proliferation because of the poor results from recent clinical trials of retinoic acid in the malignant brain tumor. Retinoic acid promoted the diffferentiation and apoptosis of tumor cell so that its effect might be obvious in the pediatric brain tumor. Therefore we are going to confirm the effectiveness of retinoic acid to inhibit the proliferation of the tumor cells; glioblastoma and medulloblastoma in childhood. Methods : Medulloblastoma cells were derived from the primary culture of the patient's specimen, and glioblastoma cells were cell lines of 373-MG and 87-MG. We estimated growth inhibition rate of each tumor cells using MTT assay in th concentration from 10¯¹²M to 10¯5M of all-trans and 13-cis retinoic acid. Results : 13-cis retinoic acid in the concentration of 10¯6M inhibited cell growth rate 10-22% on the 4th day of incubation, 10% on the 7th day, and 0-12% on the 14th day in the concentration from 10¯6M to 10¯5M. All-trans retinoic acid inhibited cell growth rate less than 5% in the concentration less than 10¯5M though the whole incubation period, but 42% on the 4th day, 37% on the 7th day, and 0% on the 14th in the concentration of 10¯5M. 13-cis retinoic acid inhibited cell growth rate 30% on the 4th day, 20% on the 7th and 14th day in the concentration between 10¯6M and 10¯5M. All-trans retinoic acid inhibited cell growth rate less than 25% by all-trans and 13-cis retnoic acid in the concentration less than 10¯6M. 13-cis retinoic acid showed 25% growth inhibition in the concentration above 10¯6M, but all-trans retnoic acid showed 40% growht inhibition in the same concentration. Conclusion : We could not find the effect of retnoic acid in the glioblastoma cells due to variable responses of the tumor cell growht inhibition in the concentration of maximum tolerable dose. IIowever, there is a significant inhibition effect of medulloblastoma cell proliferation both in the 13-cis and all-trans retinoic acid.

Mesenchymal Stem-Like Cells Derived from the Ventricle More Effectively Enhance Invasiveness of Glioblastoma Than Those Derived from the Tumor

Junseong Park,이동규,심진경,윤선진,문주형,김의현,장종희,이수재,강석구 연세대학교의과대학 2023 Yonsei medical journal Vol.64 No.3

Purpose: Glioblastoma (GBM) is one of the most lethal human tumors with a highly infiltrative phenotype. Our previous studies showed that GBM originates in the subventricular zone, and that tumor-derived mesenchymal stem-like cells (tMSLCs) promote the invasiveness of GBM tumorspheres (TSs). Here, we extend these studies in terms of ventricles using several types of GBM pa tient-derived cells. Materials and Methods: The invasiveness of GBM TSs and ventricle spheres (VSs) were quantified via collagen-based 3D invasion assays. Gene expression profiles were obtained from microarray data. A mouse orthotopic xenograft model was used for in vivo ex periments. Results: After molecular and functional characterization of ventricle-derived mesenchymal stem-like cells (vMSLCs), we investi gated the effects of these cells on the invasiveness of GBM TSs. We found that vMSLC-conditioned media (CM) significantly accel erated the invasiveness of GBM TSs and VSs, compared to the control and even tMSLC-CM. Transcriptome analyses revealed that vMSLC secreted significantly higher levels of several invasiveness-associated cytokines. Moreover, differentially expressed genes between vMSLCs and tMSLCs were enriched for migration, adhesion, and chemotaxis-related gene sets, providing a mechanistic basis for vMSLC-induced invasion of GBM TSs. In vivo experiments using a mouse orthotopic xenograft model confirmed vMSLC induced increases in the invasiveness of GBM TSs. Conclusion: Although vMSLCs are non-tumorigenic, this study adds to our understanding of how GBM cells acquire infiltrative features by vMSLCs, which are present in the region where GBM genesis originates.