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ATP-binding cassette transporters at the zebrafish blood-brain barrier and the potential utility of the zebrafish as an in vivo model

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Cancer Drug Resistance 2021年第3期4卷 620-633页

作者： Jordan M.Hotz Joanna R.Thomas Emily N.Katz Robert W.Robey Sachi Horibata Michael M.Gottesman Laboratory of cell biology Center for Cancer ResearchNational Cancer InstituteNational Institutes of HealthBethesdaMD 20892USA Zebrafish Core Eunice Kennedy Shriver National Institute of Child Health and Human DevelopmentBethesdaMD 20892USA

The brain is protected from toxins by a tightly regulated network of specialized cells,including endothelial cells,pericytes,astrocyes,and neurons,known collectively as the blood-brain barrier(BBB).This selectively permeable barrier permits only the most crucial molecules essential for brain function to enter and employs a number of different mechanisms to prevent the entry of potentially harmful toxins and *** addition to a physical barrier comprised of endothelial cells that form tight junctions to restrict paracellular transport,there is an active protective mechanism made up of energy-dependent transporters that efflux compounds back into the *** of these ATP-binding cassette(ABC)transporters are highly expressed at the BBB:P-glycoprotein(P-gp,encoded by the ABCB1 gene)and ABCG2(encoded by the ABCG2 gene).Although a number of in vitro and in vivo systems have been developed to examine the role that ABC transporters play in keeping compounds out of the brain,all have inherent advantages and ***(Danio rerio)have become a model of interest for studies of the BBB due to the similarities between the zebrafish and mammalian BBB *** this review,we discuss what is known about ABC transporters in zebrafish and what information is still needed before the zebrafish can be recommended as a model to elucidate the role of ABC transporters at the BBB.

关键词： ABC transporters blood-brain barrier ABCG2 P-glycoprotein zebrafish

An updated review of gastric cancer in the next-generation sequencing era:Insights from bench to bedside and vice versa

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World Journal of Gastroenterology 2014年第14期20卷 3927-3937页

作者： Hiroyuki Yamamoto Yoshiyuki Watanabe Tadateru Maehata Ryo Morita Yoshihito Yoshida Ritsuko Oikawa Shinya Ishigooka Shun-ichiro Ozawa Yasumasa Matsuo Kosuke Hosoya Masaki Yamashita Hiroaki Taniguchi Katsuhiko Nosho Hiromu Suzuki Hiroshi Yasuda Yasuhisa Shinomura Fumio Itoh Division of Gastroenterology and Hepatology Department of Internal MedicineSt. Marianna University School of Medicine Division of Cancer cell Research Institute of Medical ScienceUniversity of Tokyo Department of Gastroenterology Rheumatology and Clinical ImmunologySapporo Medical University School of Medicine Department of Molecular biology Sapporo Medical University School of Medicine

Gastric cancer(GC)is one of the most common malignancies and remains the second leading cause of cancer-related death *** is an increasing understanding of the roles that genetic and epigenetic alterations play in *** studies using nextgeneration sequencing(NGS)have revealed a number of potential cancer-driving genes in ***-exome sequencing of GC has identified recurrent somatic mutations in the chromatin remodeling gene ARID1A and alterations in the cell adhesion gene FAT4,a member of the cadherin gene *** in chromatin remodeling genes(ARID1A,MLL3 and MLL)have been found in 47%of ***-genome sequencing and whole-transcriptome sequencing analyses have also discovered novel alterations in *** studies of cancer epigenetics have revealed widespread alterations in genes involved in the epigenetic machinery,such as DNA methylation,histone modifications,nucleosome positioning,noncoding RNAs and *** advances in molecular research on GC have resulted in the introduction of new diagnostic and therapeutic strategies into clinical *** antihuman epidermal growth receptor 2(HER2)antibody trastuzumab has led to an era of personalized therapy in *** addition,ramucirumab,a monoclonal antibody targeting vascular endothelial growth factor receptor(VEGFR)-2,is the first biological treatment that showed survival benefits as a single-agent therapy in patients with advanced GC who progressed after firstline *** NGS to systematically identify gene alterations in GC is a promising approach with remarkable potential for investigating the pathogenesis of GC and identifying novel therapeutic targets,as well as useful *** this review,we will summarize the recent advances in the understanding of the molecular pathogenesis of GC,focusing on the potential use of these genetic and epigenetic alterations as diagnostic biomarkers and novel therapeutic targets.

关键词： Next-generation sequencing Microsatellite instability MicroRNA Epigenetic field defect Gastric washes Insulin-like growth factor 1 receptor

Corrigendum to“SOX4 contributions to TGF-b-induced endothelial-mesenchymal transition and stem cell characteristics of gastric cancer cells”[Genes&Diseases 5(2018)49-61]

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Genes & Diseases 2021年第6期8卷 946-948页

作者： Xudong Peng Guangyi Liu Hongxia Peng Anqi Chen Lang Zha Ziwei Wang Gastrointestinal Surgical unit The First Affiliated Hospital of Chongqing Medical UniversityChongqing400000PR China Department of General Surgery The First People’s HospitalYibinSichuan Province644000PR China

We regret that an error was made in“SOX4 contributions to TGF-b-induced endothelial-mesenchymal transition and stem cell characteristics of gastric cancer cells”(Genes&Diseases(2018)5,49e61).In this manuscript,the expression of SOX4 of cancer and adjacent tissues in 84 patients were detected,and 8 pairs of typical pictures were listed in Figure *** note that we made an unintentional *** confused some pictures(non-cancer tissues of NO.2 and 3)of Figure 1A due to the large number of IHC samples and pictures.

关键词： cancer Diseases Figure

Sphingosine-1-phosphate signaling in vasculogenesis and angiogenesis

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World Journal of Biological Chemistry 2010年第10期1卷 291-297页

作者： Kelley M Argraves Brent A Wilkerson W Scott Argraves Department of Regenerative Medicine and cell biology Medical University of South Carolina173 Ashley Avenue CharlestonSC 29425United States

Blood vessels either form de novo through the process of vasculogenesis or through angiogenesis that involves the sprouting and proliferation of endothelial cells in pre-existing blood vessels. A complex interactive network of signaling cascades downstream from at least three of the nine known G-protein-coupled sphingosine-1-phosphate (S1P) receptors act as a prime effector of neovascularization that occurs in embryonic development and in association with various pathologies. This review focuses on the current knowledge of the roles of S1P signaling in vasculogenesis and angiogenesis, with particular emphasis on vascular cell adhesion and motility responses.

关键词： Sphingosine-1-phosphate Vasculogenesis Angiogenesis G-protein-coupled receptors Endothelium

Ontogeny of the Hematopoietic and Vascular System:Conservation from Fish to Human

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Ontogeny of the Hematopoietic and Vascular System:Conservati...

HLA-G inhibits xenogenetic cytotoxicity mediated by human NK cells and T lymphocytes against PECs

“基因、进化与生理功能多样性”海内外学术研讨会暨中国生理学会第七届比较生理学学术会议

作者： Gustavo Gomez 林硕 Department of Molecular Cell & Developmental BiologyUniversity of CaliforniaLos AngelasLos AngelesCA 90095-1606USA Key Laboratory of cell Proliferation and Differentiation Center of Developmental Biology and GeneticsCollege of Life SciencesPeking UniversityMinistry of EducationBeijing Department of Molecular Cell & Developmental BiologyUniversity of CaliforniaLos AngelesLos AngelesCA 90095-1606USA

The ets1-related protein(etsrp) was initially identified in our lab as key gene required for endothelial development in zebrafish (Sumanas et al.,2005).Recent work by others and our lab demonstrated that etsrp acted at a very early stage to promote endothelial (Liu and Patient,2008;Pham et al., 2007;Sumanas and Lin,2006) and hematopoietic development(Liu and Patient,2008;Sumanas et al.,2008).The mouse homolog of etsrp,Er71,was also identified(Sumanas et al.,2008),whose loss-of-function led to a severe vascular and haematopoietic defect and gain-of-function converts embryonic stem cells into vascular and hematopoietic lineages(Lee et al.,2008).Similarly,human Er71/Etv2 can promote human ES cell differentiate into vascular and hematopoietic lineages.

关键词： Ontogeny VEGF signaling Mesodermal progenitors

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Chinese Science Bulletin 2003年第2期48卷 148-153页

作者： SHI Bin YIN Huijun HUANG Xiuying SUN Fangzhen Institute of Genetics and Developmental biology Chinese Academy of SciencesBeijing 100080China

In order to investigate whether the non-classi- cal HLA-G classⅠmolecule protects the porcine endothelial cells (PECs) from the lysis mediated by human immune cells in pig to human discordant xenotransplantation, we have cloned HLA-G cDNA from a human placenta by RT-PCR. Mammalian expression vector, pEFG-neo, was constructed by insertion of HLA-G cDNA in pEF-neo. We obtained effi-ciently expressed PECs by stable transfection. Cytotoxicity assay showed that overexpression of HLA-G on PECs was sufficient to inhibit human NK-92 cell lysis. The level of lysis was equal to or less than that of the lysis of human umbilical vein endothelial cells mediated by human NK-92 cells. It also indicated that HLA-G inhibited the lysis of PECs mediated by xeno-antigen specific T lymphocytes. The reduction of lysis ranged between 59.1% and 88.9%. These findings sug-gest that the transgenic approach to overexpress HLA-G is believed to be a new immunotherapy in overcoming the im-mune rejections in xenotransplantion, including delayed xenograft rejection and cell-mediated rejection.

关键词： HLA-G xenotrasplantation immune rejection gene overexpression cytotoxicity

Antitumor and antiangiogenic activities of anti-vascular endothelial growth factor hairpin ribozyme in human hepatocellular carcinoma cell cultures and xenografts

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World Journal of Gastroenterology 2007年第47期13卷 6425-6432页

作者： Li-Hua Li Zi-Jian Guo Ling-Ling Yan Ji-Cheng Yang Yu-Feng Xie Wei-Hua Sheng Zhao-Hui Huang Xue-Hao Wang Oncology Institute of Wuxi the Fourth Affiliated Hospital of Soochow University 200 Huihe Road Wuxi 214062 Jiangsu Province China Department of cellular and Molecular biology Medical School Soochow University Soochow 215007 Jiangsu Province China Department of Liver and Biliary Duct Surgery the First Affiliated Hospital of Nanjing Medical University Nanjing 210029 Jiangsu Province China

AIM: To study the effectiveness and mechanisms of anti-human vascular endothelial growth factor (hVEGF) hairpin ribozyme on angiogenesis,oncogenicity and tumor growth in a hepatocarcinoma cell line and a xenografted model. METHODS: The artificial anti-hVEGF hairpin ribozyme was transfected into hepatocarcinoma cell line SMMC-7721 and,subsequently,polymerase chain reaction (PCR) and reverse transcription polymerase chain reaction (RT-PCR) were performed to confirm the ribozyme gene integration and transcription. To determine the effects of ribozyme ,VEGF expression was detected by semiquantitative RT-PCR and enzyme liked immunosorbent assay (ELISA). MTT assay was carried out to measure the cell proliferation. Furthermore,the transfected and control cells were inoculated into nude mice respectively,the growth of cells in nude mice and angiogenesis were observed. RESULTS: VEGF expression was down-regulated sharply by ribozyme in transfected SMMC-7721 cells and xenografted tumor. Compared to the control group,the transfected cells grew slower in cell cultures and xenografts,and the xenograft formation was delayed as well. In addition,the microvessel density of the xenografted tumor was obviously declined in the transfected group. As demonstratedby microscopy,reduction of VEGF production induced by ribozyme resulted in a significantly higher cell differentiation and less proliferation vigor in xenografted tumor. CONCLUSION: Anti-hVEGF hairpin ribozyme can effectively inhibit VEGF expression and growth of hepatocarcinoma in vitro and in vivo. VEGF is functionally related to cell proliferation,differentiation and tumori-genesis in hepatocarcinoma.

关键词： Vascular endothelial growth factor Angiogenesis Hairpin ribozyme Hepatocarcinoma Gene therapy

A neurovascular unit-on-a-chip:culture and differentiation of human neural stem cells in a three-dimensional microfluidic environment

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Neural Regeneration Research 2022年第10期17卷 2260-2266页

作者： Wen-Juan Wei Ya-Chen Wang Xin Guan Wei-Gong Chen Jing Liu Stem cell Clinical Research Center National Joint Engineering LaboratoryRegenerative Medicine Centerthe First Affiliated Hospital of Dalian Medical UniversityDalianLiaoning ProvinceChina Dalian Innovation Institute of Stem cell and Precision Medicine DalianLiaoning ProvinceChina

Biological studies typically rely on a simple monolayer cell culture,which does not reflect the complex functional characteristics of human tissues and organs,or their real response to external *** technology has advantages of high-throughput screening,accurate control of the fluid velocity,low cell consumption,long-term culture,and high *** combining the multipotential differentiation of neural stem cells with high throughput and the integrated characteristics of microfluidic technology,an in vitro model of a functionalized neurovascular unit was established using human neural stem cell-derived neurons,astrocytes,oligodendrocytes,and a functional microvascular *** model comprises a multi-layer vertical neural module and vascular module,both of which were connected with a syringe *** provides controllable conditions for cell inoculation and nutrient supply,and simultaneously simulates the process of ischemic/hypoxic injury and the process of inflammatory factors in the circulatory system passing through the blood-brain barrier and then acting on the nerve tissue in the *** in vitro functionalized neurovascular unit model will be conducive to central nervous system disease research,drug screening,and new drug development.

关键词： (neural)differentiation astrocyte blood-brain barrier brain microvascular endothelial cells central nervous system microfluidics neural stem cells neuron neurovascular unit oligodendrocyte organ-on-a-chip

Generation of rat blood vasculature and hematopoietic cells in rat-mouse chimeras by blastocyst complementation

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Journal of Genetics and Genomics 2020年第5期47卷 249-261页

作者： Xiaomin Wang Hui Shi Juanjuan Zhou Qingjian Zou Quanjun Zhang Shixue Gou Pengfei Chen Lisha Mou Nana Fan Yangyang Suo Zhen Ouyang Chengdan Lai Quanmei Yan Liangxue Lai CAS Key Laboratory of Regenerative biology Guangdong Provincial Key Laboratory of Stem Cell and Regenerative MedicineSouth China Institute for Stem Cell Biology and Regenerative MedicineGuangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou510530China University of Chinese Academy of Sciences Beijing100049China Guangzhou Regenerative Medicine and Health Guangdong Laboratory(GRMH-GDL) Guangzhou510005China Institute for Stem cell and Regeneration Chinese Academy of SciencesBeijing100101China Research unit of Generation of Large Animal Disease Models Chinese Academy of Medical Sciences(2019RU015)Guangzhou510530China School of Biotechnology and Health Sciences Wuyi UniversityJiangmen529020China Shenzhen Xenotransplantation Medical Engineering Research and Development Center Institute of Translational MedicineShenzhen University Health Science CenterShenzhen University School of MedicineFirst Affiliated Hospital of Shenzhen UniversityShenzhen Second People's HospitalShenzhen518035China Department of Central Laboratory Shenzhen Longhua District Central HospitalShenzhen518110China School of Life Sciences University of Science and Technology of ChinaHefei230027China Joint School of Life Sciences Guangzhou Institutes of Biomedicine and HealthChinese Academy of SciencesGuangzhou Medical UniversityGuangzhou511436China

Interspecies chimera through blastocyst complementation could be an alternative approach to create human organs in animals by using human pluripotent stem cells.A mismatch of the major histocompatibility complex of vascular endothelial cells between the human and host animal will cause graft rejection in the transplanted ***,to achieve a transplantable organ in animals without rejection,creation of vascular endothelial cells derived from humans within the organ is *** this study,to explore whether donor xeno-pluripotent stem cells can compensate for blood vasculature in host animals,we generated rat-mouse chimeras by injection of rat embryonic stem cells(rESCs)into mouse blastocysts with deficiency of Flk-1 protein,which is associated with endothelial and hematopoietic cell *** found that rESCs could differentiate into vascular endothelial and hematopoietic cells in the rat-mouse *** whole yolk sac(YS)of Flk-1^EGFP/ECFP rat-mouse chimera was full of rat blood *** genes related to vascular endothelial cells,arteries,and veins,blood vessels formation process,as well as hematopoietic cells,were highly expressed in the *** results suggested that rat vascular endothelial cells could undergo proliferation,migration,and self-assembly to form blood vasculature and that hematopoietic cells could differentiate into B cells,T cells,and myeloid cells in rat-mouse chimeras,which was able to rescue early embryonic lethality caused by Flk-1 deficiency in mouse.

关键词： Blastocyst complementation Interspecies chimera Intraspecies chimera Flk-1 Vascular endothelial cell Hematopoietic cell