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Mesenchymal stem cells and the neuronal microenvironment in the area of spinal cord injury

Neural Regeneration Research 2019年第2期14卷 227-237页

作者： Yana O.Mukhamedshina Olga A.Gracheva Dina M.Mukhutdinova Yurii A.Chelyshev Albert A.Rizvanov OpenLab Gene and Cell Technologies Institute of Fundamental Medicine and Biology Kazan (Volga Region)Federal University Department of Histology Cytology and Embryology Kazan State Medical University Department of Therapy and Clinical Diagnostics with radiology Faculty of Veterinary Medicine Bauman Kazan State Academy of Veterinary Medicine OpenLab Gene and Cell Technologies Institute of Fundamental Medicine and Biology Kazan (Volga Region) Federal University

cell-based technologies are used as a therapeutic strategy in spinal cord injury(SCI). Mesenchymal stem cells(MSCs), which secrete various neurotrophic factors and cytokines, have immunomodulatory, anti-apoptotic and anti-inflammatory effects, modulate reactivity/phenotype of astrocytes and the microglia, thereby promoting neuroregeneration seem to be the most promising. The therapeutic effect of MSCs is due to a paracrine mechanism of their action, therefore the survival of MSCs and their secretory phenotype is of particular importance. Nevertheless, these data are not always reported in efficacy studies of MSC therapy in SCI. Here, we provide a review with summaries of preclinical trials data evaluating the efficacy of MSCs in animal models of SCI. Based on the data collected, we have tried(1) to establish the behavior of MSCs after transplantation in SCI with an evaluation of cell survival, migration potential, distribution in the area of injured and intact tissue and possible differentiation;(2) to determine the effects MSCs on neuronal microenvironment and correlate them with the efficacy of functional recovery in SCI;(3) to ascertain the conditions under which MSCs demonstrate their best survival and greatest efficacy.

关键词： spinal cord injury mesenchymal stem cells survival migration Rho/ROCK/PTEN astrocytes microglia myelin-forming cells axon growth tissue integrity

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Angiogenesis and nerve regeneration induced by local administration of plasmid pBud-coVEGF165-coFGF2 into the intact rat sciatic nerve

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Neural Regeneration Research 2021年第9期16卷 1882-1889页

作者： Ruslan Masgutov Alina Zeinalova Alexey Bogov Galina Masgutova Ilnur Salafutdinov Ekaterina Garanina Valeriia Syromiatnikova Kamilla Idrisova Adelya Mullakhmetova Dina Andreeva Liliya Mukhametova Adilet Kadyrov Igor Pankov Albert Rizvanov openlab“gene and cell technologies” Institute of Fundamental Medicine and BiologyKazan Federal UniversityKazanRussia Republican Clinical Hospital KazanRussia Department of Traumatology and Orthopedics Kazan State Medical AcademyKazanRussia

Vascular endothelial growth factor(VEGF) and fibroblast growth factor 2(FGF2) are well-known growth factors involved in the regeneration of various tissues and organs, including peripheral nerve system. In the present study, we elucidated the local and systemic effects of plasmid construct рBud-coVEGF165-coFGF2 injected into the epineurium of intact rat sciatic nerve. Results of histological examination of sciatic nerve and multiplex immunoassays of serum showed the absence of immunogenicity and biosafety of plasmid рBud-coVEGF165-coFGF2. Moreover, local administration of plasmid DNA construct resulted in significantly decreased levels of pro-inflammatory cytokines in the peripheral blood, including tumor necrosis factor α(TNFα) and interleukin-12, and significantly increased levels of cytokines and chemokines including Regulated upon Activation, Normal T cell Expressed and Presumably Secrete(RANTES), epidermal growth factor, interleukin-2, and monocyte chemoattractant protein 1. These changes in the peripheral blood on day 7 after injection of plasmid construct рBud-coVEGF165-coFGF2 show that the plasmid construct has systemic effects and may modulate immune response. At the same time, reverse transcriptionpolymerase chain reaction revealed transient expression of coFGF2, coVEGF165, ratFGF2 and ratVEGFA with direct transport of transcripts from distal part to proximal part of the sciatic nerve. Immunohistochemical staining revealed prolonged presence of VEGFA in sciatic nerve till 14 days post-injection. These findings suggest that local administration of plasmid construct рBud-coVEGF165-coFGF2 at a concentration of 30 ng/μL results in the formation of pro-angiogenic stimuli and, and the plasmid construct, used as a drug for gene therapy, might potentially facilitate regeneration of the sciatic nerve. The study was approved by the Animal Ethics Committee of Kazan Federal University, procedures were approved by the Local Ethics Committee(approval No. 5) on May 27, 2014.

关键词： chemokines cytokines fibroblast growth factor gene therapy growth factors peripheral nerve system sciatic nerve regeneration vascular endothelial growth factor

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An improved U-Net for cell confluence estimation

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Optoelectronics Letters 2022年第6期18卷 378-384页

作者： BAI Hua LU Changhao MA Ming YAN Shulin ZHANG Jianzhong HAN Zhibo Tianjin Key Laboratory of Optoelectronic Detection Technology and Systems School of Electronic and Information EngineeringTiangong UniversityTianjin300387China Tianjin Key Laboratory of Engineering technologies for cell Phamaceutical Tianjin 300457China National Engineering Research Center of cell Products/Am cell gene Co. Ltd.Tianjin 300457China State Key Lab of Experimental Hematology Chinese Academy of Medical Sciences&Peking Union Medical CollegeTianjin 300020China

cell confluence is an important metric to determine the growth and the best harvest time of adherent cells. At present, the evaluation of cell confluence mainly relies on experienced labor, and thus it is not conducive to the automated cell culture. In this paper, we proposed an improved U-Net algorithm(called DU-Net) for the segmentation of adherent cells. First, the general convolution was replaced by the dilated convolution to expand the receptive fields for feature extraction. Then, the convolutional layers were combined with the batch normalization layers to reduce the dependence of the network on initialization. As a result, the segmentation accuracy and F1-score of the proposed DU-Net for adherent cells with low confluence(<50%) reached 96.94% and 93.87%, respectively, and for those with high confluence(≥50%), they reached 98.63% and 98.98%, respectively. Further, the paired t-test results showed that the proposed DU-Net was statistically superior to the traditional U-Net algorithm.

关键词： convolution estimation batch

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miR-137 inhibits melanoma cell proliferation through downregulation of GLO1

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Science China(Life Sciences) 2018年第5期61卷 541-549页

作者： Na Lv Shuai Hao Chonglin Luo Alia Abukiwan Ying Hao Fei Gai Weiwei Huang Lingyun Huang Xueyuan Xiao Stefan B.Eichmüller Dacheng He Key Laboratory of cell Proliferation and Regulation of Ministry of Education Universities of the Confederated Institute for ProteomicsBeijing Normal University Beijing Engineering and Technology Research Center of Food Additives Beijing Technology and Business University gene & Care Health technologies Co. Ltd. GMP & T cell Therapy Unit German Cancer Research Center (DKFZ) College of Sports Northwest Normal University

Late-stage melanoma is refractory to current therapies. MicroRNAs (miRNAs) can modulate many physiological and pathological processes of melanoma. Studies have demonstrated that miR-137 acts as a tumor suppressor by inhibiting the proliferation of melanoma cells through targeting multiple mRNAs. The glyoxalase system member glyoxalase 1 (GLO1) is the principal scavenging enzyme of methylglyoxal (MG), a toxic byproduct of glycolysis. Using 35S in vivo/vitro labelling analysis for dynamic proteomics (SiLAD), we found that miR-137 downregulated the expression of GLO1 in melanoma cells.Bioinformatics analysis predicted that GLO1 is a direct target of miR-137. This was validated by dual luciferase reporter assay.Quantitative RT-PCR (qRT-PCR) and western blot analysis indicated that miR-137 could decrease endogenous GLO1 expression.Furthermore, siRNA targeting of GLO1 mimicked inhibition of melanoma cell proliferation caused by miR-137 overexpression.Re-expression of GLO1 was able to restore miR-137-mediated suppression of melanoma cell proliferation. Therefore, these results suggest that miR-137 inhibits the proliferation of melanoma cells by targeting GLO1.

关键词：房间 proteomics RT-PCR siRNA 病理学指向副产品 35S

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Spontaneous xenogeneic GvHD in Wilms'tumor Patient-Derived xenograft models and potential solutions

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Animal Models and Experimental Medicine 2022年第4期5卷 389-396页

作者： Seyed Mostafa Monzavi Ahad Muhammadnejad Maryam Behfar Amir Arsalan Khorsand Samad Muhammadnejad Abdol-Mohammad Kajbafzadeh Department of Applied cell Sciences School of Advanced Technologies in MedicineTehran University of Medical SciencesTehranIran Pediatric Urology and Regenerative Medicine Research Center Tehran University of Medical SciencesTehranIran gene Therapy Research Center Digestive Diseases Research InstituteTehran University of Medical SciencesTehranIran Cancer Biology Research Center Cancer Institute of IranTehran University of Medical SciencesTehranIran PDX Platform Biomarker Evaluation and Supervision Team for Personalized MedicineMolecular Tumor BoardCancer Institute of IranTehran University of Medical SciencesTehranIran Pediatric cell and gene Therapy Research Center Tehran University of Medical SciencesTehranIran

Severely immunocompromised NOD.Cg-PrkdcIl2rg(NOG)mice are among the ideal animal recipients for generation of human cancer models.Transplantation of human solid tumors having abundant tumor-i nfiltrating lymphocytes(TILs)can induce xenogeneic graft-versus-host disease(xGvHD)following engraftment and expansion of the TILs inside the animal body.Wilms’tumor(WT)has not been recognized as a lymphocyte-predominant tumor.However,3 consecutive generations of NOG mice bearing WT patient-derived xenografts(PDX)xenotransplanted from a single donor showed different degrees of inflammatory symptoms after transplantation before any therapeutic intervention.In the initial generation,dermatitis,auto-amputation of digits,weight loss,lymphadenopathy,hepatitis,and interstitial pneumonitis were observed.Despite antibiotic treatment,no response was noticed,and thus the animals were prematurely euthanized(day 47 posttransplantation).Laboratory and histopathologic evaluations revealed lymphoid infiltrates positively immunostained with anti-human CD3 and CD8 antibodies in the xenografts and primary tumor,whereas no microbial infection or lymphoproliferative disorder was found.Mice of the next generation that lived longer(91 days)developed sclerotic skin changes and more severe pneumonitis.Cutaneous symptoms were milder in the last generation.The xenografts of the last 2 generations also contained TILs,and lacked lymphoproliferative transformation.The systemic immunoinflammatory syndrome in the absence of microbial infection and posttransplant lymphoproliferative disorder was suggestive of xGvHD.While there are few reports of xGvHD in severely immunodeficient mice xenotransplanted from lymphodominant tumor xenografts,this report for the first time documented serial xGvHD in consecutive passages of WT PDX-bearing models and discussed potential solutions to prevent such an undesired complication.

关键词： graft-versus-host disease patient-derived xenograft models tumor-infiltrating lymphocytes Wilms’tumor

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Cancer immunotherapy: an evolving paradigm

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2022年第10期23卷 791-792页

作者： Aifu LIN MOE Laboratory of Biosystem Homeostasis and Protection College of Life SciencesZhejiang UniversityHangzhou 310058China Cancer Center Zhejiang UniversityHangzhou 310058China Key Laboratory for cell and gene Engineering of Zhejiang Province Hangzhou 310058China

The inhibition of the host’s natural immune response by tumor cells was widely reported in the early phases of the development of oncology therapy,and the concept of employing the host’s immune system to treat cancer,i.***.tumor immunotherapy,is not new.However,as a result of early theoretical constraints,clinical application of immunotherapy did not go smoothly and lagged significantly behind radiation and chemotherapy.The path has been winding,but the future now seems promising.Immunotherapy research has advanced enormously as a result of the maturing of immuno-editing theory and the creation of numerous technologies,despite a number of unsuccessful endeavors and clinical studies.Since around 1998,the US Food and Drug Administration(FDA)has approved a variety of tumor immunotherapies,including cytokines(interleukin-2,interferons),cancer vaccines(Provenge),immune checkpoint inhibitors(ipilimumab),and cellular therapies(chimeric antigen receptor-T(CAR-T)),signaling a boom in the field.

关键词： immunotherapy clinical chemotherapy

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Technological breakthroughs in generating transgene-free and genetically stable CRISPR-edited plants

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aBIOTECH 2020年第1期1卷 88-96页

作者： Yubing He Yunde Zhao National Key Laboratory of Crop genetic Improvement and National Center of Plant gene Research(Wuhan) Huazhong Agricultural UniversityWuhan 430070China Section of cell and Developmental Biology University of California San DiegoLa JollaCA 92093-0116USA

CRISPR/Cas9 gene-editing technologies have been very effective in editing target genes in all major crop plants and offer unprecedented potentials in crop improvement.A major challenge in using CRISPR gene-editing technology for agricultural applications is that the target gene-edited crop plants need to be transgene free to maintain trait stability and to gain regulatory approval for commercial production.In this article,we present various strategies for generating transgene-free and target geneedited crop plants.The CRISPR transgenes can be removed by genetic segregation if the crop plants are reproduced sexually.Marker-assisted tracking and eliminating transgenes greatly decrease the time and labor needed for identifying the ideal transgene-free plants.Transgenes can be programed to undergo self-elimination when CRISPR genes and suicide genes are sequentially activated,greatly accelerating the isolation of transgene-free and target gene-edited plants.Transgene-free plants can also be generated using approaches that are considered non-transgenic such as ribonucleoprotein transfection,transient expression of transgenes without DNA integration,and nano-biotechnology.Here,we discuss the advantages and disadvantages of the various strategies in generating transgene-free plants and provide guidance for adopting the best strategies in editing a crop plant.

关键词： CRISPR Transgene-free Marker-assisted selection TKC Cas9 gene editing

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用合成免疫学打开肿瘤免疫治疗的瓶颈(英文)

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Science Bulletin 2015年第11期60卷 977-985,I0007页

作者：陈志英马飞黄海亮何成宜 Laboratory for gene and cell Therapy Chinese Academy of Sciences Shenzhen Institute of Advanced Technology Analytic and Translational genetics Unit Massachusetts General Hospital Broad Institute of Massachusetts Institute of Technology and Harvard University

近年来,肿瘤免疫治疗取得了令人瞩目的重大突破,采用过继免疫技术输入工程抗体或工程效应T细胞,一些接受治疗的患者获得持久的缓解甚至痊愈.然而,这些单靶点治疗技术对部分患者没有疗效.有证据表明,肿瘤的异质性(heterogeneity)和不断进... 详细信息

近年来,肿瘤免疫治疗取得了令人瞩目的重大突破,采用过继免疫技术输入工程抗体或工程效应T细胞,一些接受治疗的患者获得持久的缓解甚至痊愈.然而,这些单靶点治疗技术对部分患者没有疗效.有证据表明,肿瘤的异质性(heterogeneity)和不断进化(evolving)的特性是造成治疗失败的原因,只有发展出针对这些肿瘤特性的新方法,才能实现新的突破.本文提出用合成免疫策略(synthetic immunity,SI)来实现这一目标.SI的原则是通过制备一组工程抗体和抗体重新定向的细胞毒T淋巴细胞(antibody-retargeted cytotoxic T cells)高度特异性地杀死全部肿瘤细胞系而达到根治的目的.要实现这一目标,只能通过系统化方法来选择组合工程抗体,具体是建立一个肿瘤抗原系统树(tumor epitope-tree)来确定肿瘤细胞系及其抗原的系统构成,然后选择"树干或主要分杈"上的"抗原靶点组"来进行过继免疫治疗,最终消灭所有肿瘤细胞系.同时结合其他技术如扩大抗原表位数量、提高抗体亲和力和T细胞活性等,可以进一步提高SI的效率.此外,用非病毒DNA基因载体在体内生产工程抗体将是一个安全、有效和经济的解决方案.

关键词：免疫治疗抗肿瘤治疗效果抗体生成抗原表位出租车免疫疗法诱导免疫

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Complexities and pitfalls in analyzing and interpreting mitochondrial DNA content in human cancer

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Journal of genetics and Genomics 2020年第7期47卷 349-359页

作者： Hieu Nguyen Thomas LaFramboise Vinmec Research Institute of Stem cell and gene Technology(VRSIG) 458 M inh KhaiVinh TuyHai Ba TrungHanoiViet Nam Department of genetics and Genome Sciences Case Western Reserve University School of Medicine10900 Euclid AvenueClevelandOH44106USA

Mutations in the human mitochondrial genome have been observed in all types of human cancer,indicating that mutations might contribute to tumorigenesis,metastasis,recurrence,or drug response.This possibility is appealing because of the known shift from oxidative metabolism to glycolysis,known as the Warburg effect,that occurs in malignancy.Mitochondrial DNA(mtDNA)mutations could either bematernally inherited and predispose to cancer(germ line mutations)or occur sporadically in the mtDNA of specific tissues(rissue-or tumor-specific somatic mutations)and contribute to the tumor initiation and progression process.High-throughput sequencing technologies now enable comprehensive detection of mtDNA variation in tissues and bodily fluids,with the potential to be used as an early detectiontool that may impact the treatment of cancer.Here,we discuss insights into the roles of mtDNA mutations in carcinogenesis,highlighting the complexities involved in the analysis and interpretation of mitochondrial genomic content.technical challenges in studying their contribution to pathogenesis,and the value of mtDNA mutations in developing early detection,diagnosis,prognosis,and therapeuticstrategics for cancer.

关键词： Mitochondrial DNA Cancer genetic variation

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The applications of induced pluripotent stem(iPS)cells in drug development

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Frontiers in Biology 2011年第1期6卷 52-57页

作者： Shulong YANG Xuelian WANG Jinmiao LIU Zhao LIU Jiaxue HUANG Life technologies Vcanland Holding Group Union Stem cell&gene Engineering Co.LTD Tianjin 300384China School of Pharmaceutical Science and Technology Tianjin UniversityTianjin 300072China

The introduction of induced pluripotent stem(iPS)cells has been a milestone in the field of regenerative medicine and drug discovery.iPS cells can provide a continuous and individualized source of stem cells and are considered to hold great potential for economically feasible personalized stem cell therapy.Various diseases might potentially be cured by iPS cell-based therapy including Parkinson’s disease,Alzheimer’s disease,Huntington disease,ischemic heart disease,diabetes and so on.Moreover,iPS cells derived from patients suffering from unique incurable diseases can be developed into patient-and disease-specific cell lines.These cells can be used as an effective approach to study the mechanisms of diseases,providing useful tools for drug discovery,development and evaluation.The development of suitable methods for the culture and expansion of iPS cells and their differentiated progenies make feasible modern drug discovery techniques such as high-throughput screening.Furthermore,iPS cells can be applied in the field of toxicological and pharmacokinetics tests.This review focuses on the applications of iPS cells in the field of pharmaceutical industry.

关键词： induced pluripotent stem(iPS)cells drug discovery

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