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Improvement of histone deacetylase inhibitor efficacy by SN38 through TWIST1 suppression in synovial sarcoma

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Cancer Innovation 2024年第2期3卷 80-95页

作者： Satoru Sasagawa Jun Kumai Toru Wakamatsu Yoshihiro Yui Molecular Biology Laboratory Research InstituteNozaki Tokushukai HospitalDaitoOsakaJapan Sarcoma Treatment Laboratory Research InstituteNozaki Tokushukai HospitalDaitoOsakaJapan Department of Musculoskeletal Oncology Service Osaka International Cancer InstituteOsakaJapan

Background:Synovial sarcoma(SS)is an SS18-SSX fusion gene-driven soft tissue sarcoma with mesenchymal characteristics,associated with a poor prognosis due to frequent metastasis to a distant organ,such as the lung.Histone deacetylase(HDAC)inhibitors(HDACis)are arising as potent molecular targeted drugs,as HDACi treatment disrupts the SS oncoprotein complex,which includes HDACs,in addition to general HDACi effects.To provide further molecular evidence for the advantages of HDACi treatment and its limitations due to drug resistance induced by the microenvironment in SS cells,we examined cellular responses to HDACi treatment in combination with two-dimensional(2D)and 3D culture conditions.Methods:Using several SS cell lines,biochemical and cell biological assays were performed with romidepsin,an HDAC1/2 selective inhibitor.SN38 was concomitantly used as an ameliorant drug with romidepsin treatment.Cytostasis,apoptosis induction,and MHC class I polypeptide-related sequence A/B(MICA/B)induction were monitored to evaluate the drug efficacy.In addition to the conventional 2D culture condition,spheroid culture was adopted to evaluate the influence of cell-mass microenvironment on chemoresistance.Results:By monitoring the cellular behavior with romidepsin and/or SN38 in SS cells,we observed that responsiveness is diverse in each cell line.In the apoptotic inducible cells,co-treatment with SN38 enhanced cell death.In nonapoptotic inducible cells,cytostasis and MICA/B induction were observed,and SN38 improved MICA/B induction further.As a novel efficacy of SN38,we revealed TWIST1 suppression in SS cells.In the spheroid(3D)condition,romidepsin efficacy was severely restricted in TWIST1-positive cells.We demonstrated that TWIST1 downregulation restored romidepsin efficacy even in spheroid form,and concomitant SN38 treatment along with romidepsin reproduced the reaction.Conclusions:The current study demonstrated the benefits and concerns of using HDACi for SS treatment in 2D and 3D culture conditions and provided molecular evi

关键词： HDAC inhibitor MICA/B SN38 spheroid synovial sarcoma TWIST1

Drag increment of internal waves generated by horizontally moving spheroid in supercritical regime

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Acta Mechanica Sinica 2008年第2期24卷 127-132页

作者： Zhaoting Xu Hao Xu V. A. Gorodtsov Physical Oceanography Laboratory and Institute of PhysicalOceanography Ocean University of China266100 Qingdao China School of Naval Architecture Ocean and Civil EngineeringShanghai Jiaotong University200030 Shanghai China Institute for Problem in Mechanics The Russian Academy of Science 119526 Moscow Russia

A theory on the drag increment of internal waves with a spheroid moving horizontally at a high velocity （or for large internal Froude number） in uniformly vertically stratified fluid （or ocean） is presented in the present paper. A surface source distribution is employed to model a hydrodynamic interaction between the spheroid and the stratified fluid. From theoretical results, it is shown that there exists an asymptote of zero-drag increment in supercritical regimes, where internal Froude numbers are larger than the critical internal Froude numbers. When the spheroid reduces to a sphere, the results in this paper is in good agreement with the previous theoretical and experimental results of the sphere.

关键词： Internal wave Stratified flow Drag increment spheroid Internal Froude number

Porous hydrogel arrays for hepatoma cell spheroid formation and drug resistance investigation

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Bio-Design and Manufacturing 2021年第4期4卷 842-850页

作者： Xin Lei Changmin Shao Xin Shou Keqing Shi Liang Shi Yuanjin Zhao Translational Medicine Laboratory The First Affiliated Hospital of Wenzhou Medical UniversityWenzhou 325035China Wenzhou Institute University of Chinese Academy of SciencesWenzhou 325001China State Key Laboratory of Bioelectronics School of Biological Science and Medical EngineeringSoutheast UniversityNanjing 210096China Department of Laboratory Medicine The Eighth Affiliated HospitalSun Yat-Sen UniversityShenzhen 518033China

Drug resistance is one of the major obstacles in the drug therapy of cancers.Efforts in this area in pre-clinical research have focused on developing novel platforms to evaluate and decrease drug resistance.In this paper,inspired by the structure of hives where swarms live and breed,we propose porous hydrogel arrays with a uniform pore structure for the generation of hepatoma cell spheroids and the investigation of drug resistance.The porous hydrogel arrays were fabricated using polyeth-ylene glycol diacrylate(PEGDA)hydrogel to negatively replicate a well-designed template.Benefiting from the elaborate processing of the template,the prepared porous hydrogel arrays possessed a uniform pore structure.Due to their anti-adhesion properties and the excellent biocompatibility of the PEGDA hydrogel,the hepatoma cells could form well-defined and uni-form hepatoma cell spheroids in the porous hydrogel arrays.We found that the resistant hepatoma cell spheroids showed more significant Lenvatinib resistance and a migratory phenotype compared with a two-dimensional(2D)cell culture,which reveals the reason for the failure of most 2D cell-selected drugs for in vivo applications.These features give such porous hydrogel arrays promising application prospects in the investigation of tumor cell spheroid culture and in vitro drug resistance.

关键词： Drug resistance Hydrogel Porous spheroid Hepatoma

Cellulose nanofibril matrix drives the dynamic formation of spheroids

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2023年第10期24卷 922-934页

作者： Yi LU Guo LI Yeqiu LI Yuan YAO College of Chemical and Biological Engineering Zhejiang UniversityHangzhou 310027China ZJU-Hangzhou Global Scientific and Technological Innovation Center Zhejiang UniversityHangzhou 311215China School of Physical Science and Technology ShanghaiTech UniversityShanghai 201210China

Multicellular spheroids,which mimic the natural organ counterparts,allow the prospect of drug screening and regenerative medicine.However,their application is hampered by low processing efficiency or limited scale.This study introduces an efficient method to drive rapid multicellular spheroid formation by a cellulose nanofibril matrix.This matrix enables the facilitated growth of spheroids(within 48 h)through multiple cell assembly into size-controllable aggregates with well-organized physiological microstructure.The efficiency,dimension,and conformation of the as-formed spheroids depend on the concentration of extracellular nanofibrils,the number of assembled cells,and the heterogeneity of cell types.The above strategy allows the robust formation mechanism of compacted tumoroids and hepatocyte spheroids.

关键词： Cellulose Nanofibril Matrix Self-assembly spheroid

Therapeutic strategies of three-dimensional stem cell spheroids and organoids for tissue repair and regeneration

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Bioactive Materials 2023年第1期19卷 50-74页

作者： Woochan Kim Yonghyun Gwon Sunho Park Hyoseong Kim Jangho Kim Department of Convergence Biosystems Engineering Chonnam National UniversityGwangju61186Republic of Korea Department of Rural and Biosystems Engineering Chonnam National UniversityGwangju61186Republic of Korea Interdisciplinary Program in IT-Bio Convergence System Chonnam National UniversityGwangju61186Republic of Korea

Three-dimensional(3D)stem cell culture systems have attracted considerable attention as a way to better mimic the complex interactions between individual cells and the extracellular matrix(ECM)that occur in vivo.Moreover,3D cell culture systems have unique properties that help guide specific functions,growth,and processes of stem cells(e.g.,embryogenesis,morphogenesis,and organogenesis).Thus,3D stem cell culture systems that mimic in vivo environments enable basic research about various tissues and organs.In this review,we focus on the advanced therapeutic applications of stem cell-based 3D culture systems generated using different engineering techniques.Specifically,we summarize the historical advancements of 3D cell culture systems and discuss the therapeutic applications of stem cell-based spheroids and organoids,including engineering techniques for tissue repair and regeneration.

关键词： 3D stem cell culture spheroid Organoid Tissue repair Tissue regeneration

Rock inhibitor may compromise human induced pluripotent stem cells for cardiac differentiation in 3D

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Bioactive Materials 2022年第3期7卷 508-522页

作者： Bin Jiang Wenquan Ou James G.Shamul Hao Chen Sarah Van Belleghem Samantha Stewart Zhenguo Liu John P.Fisher Xiaoming He Fischell Department of Bioengineering University of MarylandCollege ParkMD20742United States Division of Cardiovascular Medicine University of Missouri School of MedicineColumbiaMO65212United States Marlene and Stewart Greenebaum Comprehensive Cancer Center University of MarylandBaltimoreMD21201United States

Cardiomyocytes differentiated from human induced pluripotent stem cells(iPSCs)are valuable for the understanding/treatment of the deadly heart diseases and their drug screening.However,the very much needed homogeneous 3D cardiac differentiation of human iPSCs is still challenging.Here,it is discovered surprisingly that Rock inhibitor(RI),used ubiquitously to improve the survival/yield of human iPSCs,induces early gastrulation-like change to human iPSCs in 3D culture and may cause their heterogeneous differentiation into all the three germ layers(i.e.,ectoderm,mesoderm,and endoderm)at the commonly used concentration(10μM).This greatly compromises the capacity of human iPSCs for homogeneous 3D cardiac differentiation.By reducing the RI to 1μM for 3D culture,the human iPSCs retain high pluripotency/quality in inner cell mass-like solid 3D spheroids.Consequently,the beating efficiency of 3D cardiac differentiation can be improved to more than 95%in~7 days(compared to less than~50%in 14 days for the 10μM RI condition).Furthermore,the outset beating time(OBT)of all resultant cardiac spheroids(CSs)is synchronized within only 1 day and they form a synchronously beating 3D construct after 5-day culture in gelatin methacrylol(GelMA)hydrogel,showing high homogeneity(in terms of the OBT)in functional maturity of the CSs.Moreover,the resultant cardiomyocytes are of high quality with key functional ultrastructures and highly responsive to cardiac drugs.These discoveries may greatly facilitate the utilization of human iPSCs for understanding and treating heart diseases.

关键词： Episomal iPSC Cardiomyocyte spheroid GelMA

INVASION OF spheroid OF MURINE LUNG ADENOCARCINOMA (LA_(795)) CELLS INTO EMBRYONIC CHICK HEART FRAGMENTS

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Chinese Journal of Cancer Research 1992年第4期4卷 20-25页

作者：许三多高进 Department of Cell Biology Beijing Medical University Beijing Department of Pathology Institute of Basic Medical Sciences Chinese Academy

An in vitro spheriod invasion model of tumor cell was established by using murine lung adenocarcinoma cell line (LA795) and precultured embryonic chick heart fragment (PHF). The spheroid of LA795 cells were prepared by incubating a suspension of trypsinized LA795 cells on a gyratory shaker. spheroid aggregates of LA795 cells in diameter of 0. 2 mm were selected and confronted with PHF (diameter of 0. 4 mm) on semi- solid medium for 3 - 4 hours, then, individual confronting pain were transferred into fluid medium for further co-culture on gyratory shaker. After 1, 3, 5 and 7 days, multiple confronting pairs were processed for histological and ultrastructural study. The Invasive capacity and the invasion process of LA795 cells were examined and observed. The results demonstrated that LA795 cell line has a high capacity of invasion and high malignancy in vitro. This spheroid Invasion model is very useful for studying mechanism of Invasiveness of tumor cells in vitro.

关键词： Invasion spheroid lung adenocarcinoma precultured chick heart fragment (PHF) organ culture.

Applications of a new In vivo tumor spheroid based shell-less chorioallantoic membrane 3-D model in bioengineering research

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Journal of Biomedical Science and Engineering 2010年第1期3卷 20-26页

作者： Nzola De Magalhaes Lih-Huei L.Liaw Michael Berns Vittorio Cristini Zhongping Chen Dwayne Stupack John Lowengrub Department of Biomedical Engineering University of CaliforniaIrvineUSA Beckman Laser Institute and Medical Clinic University of CaliforniaIrvineUSA Health Sciences Center University of TexasHoustonHoustonUSA Moores Cancer Center University of CaliforniaSan DiegoUSA Department of Mathematics University of CaliforniaIrvineUSA

The chicken chorioallantoic membrane (CAM) is a classical in vivo biological model in studies of angiogenesis. Combined with the right tumor system and experimental configuration this classical model can offer new approaches to investigating tumor processes. The increase in development of biotechnolo- gical devices for cancer diagnosis and treatment, calls for more sophisticated tumor models that can easily adapt to the technology, and provide a more accurate, stable and consistent platform for rapid quantitative and qualitative analysis. As we discuss a variety of applications of this novel in vivo tumor spheroid based shell-less CAM model in biomedical engineering research, we will show that it is extremely versatile and easily adaptable to an array of biomedical applications. The model is particularly useful in quantitative studies of the progression of avascular tumors into vascularized tumors in the CAM. Its environment is more stable, flat and has a large working area and wider field of view excellent for imaging and longitudinal studies. Finally, rapid data acquisition, screening and validation of biomedical devices and therapeutics are possible with the short experimental window.

关键词： CAM Cancer spheroid Optical Coherence Tomography Photodynamic Therapy Computational Modeling Angiogenesis

Changes of protein expression during tumorosphere formation of small cell lung cancer circulating tumor cells

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Oncology Research 2023年第1期31卷 13-22页

作者： SANDRA STICKLER BARBARA RATH MAXIMILIAN HOCHMAIR CLEMENS LANG LUKAS WEIGL GERHARD HAMILTON Institute of Pharmacology Medical University of ViennaViennaAustria Karl Landsteiner Institute of Lung Research and Pulmonary Oncology Klinik FloridsdorfViennaAustria Department of Trauma Surgery Sozialmedizinisches Zentrum OstDonauspitalViennaAustria Division of Special Anesthesia and Pain Medicine Medical University of ViennaViennaAustria

Small cell lung cancer(SCLC)is frequently disseminated and has a dismal prognosis with survival times of approximately two years.This cancer responds well to initial chemotherapy but recurs within a short time as a globally chemoresistant tumor.Circulating tumor cells(CTCs)are held responsible for metastasis,the extremely high numbers of these cells in advanced SCLC allowed us to establish several permanent CTC cell lines.These CTCs are distinguished by the spontaneous formation of large spheroids,termed tumorospheres,in regular tissue culture.These contain quiescent and hypoxic cells in their interior and are associated with high chemoresistance compared to single cell cultures.Nine CTC lines were compared for their expression of 84 proteins associated with cancer either as single cells or in the form of tumorospheres in Western blot arrays.With the exception of the UHGc5 line,all other CTC lines express EpCAM and lack a complete EpCAM-negative,vimentin-positive epithelial-mesenchymal transition(EMT)phenotype.Upon formation of tumorospheres the expression of EpCAM,that mediates cell-cell adhesion is markedly upregulated.Proteins such as E-Cadherin,p27 KIP1,Progranulin,BXclx,Galectin-3,and Survivin showed variable changes for the distinct CTC cell lines.In conclusion,EpCAM presents the most critical marker for individual SCLC CTCs and the assembly of highly chemoresistant tumorospheres.

关键词： SCLC spheroid Chemoresistance EpCAM Topotecan Epirubicin