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adenovirus-mediated gene delivery:Potential applications for gene and cell-based therapies in the new era of personalized medicine

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Genes & Diseases 2017年第2期4卷 43-63页

作者： Cody S.Lee Elliot S.Bishop Ruyi Zhang Xinyi Yu Evan M.Farina Shujuan Yan Chen Zhao Zongyue Zeng Yi Shu Xingye Wu Jiayan Lei Yasha Li Wenwen Zhang Chao Yang Ke Wu Ying Wu Sherwin Ho Aravind Athiviraham Michael J.Lee Jennifer Moriatis Wolf Russell R.Reid Tong-Chuan He The University of Chicago Pritzker School of Medicine ChicagoIL 60637USA Laboratory of Craniofacial Biology and Development Section of Plastic and Reconstructive SurgeryDepartment of SurgeryThe University of Chicago Medical CenterChicagoIL 60637USA Molecular Oncology Laboratory Department of Orthopaedic Surgery and Rehabilitation MedicineThe University of Chicago Medical CenterChicagoIL 60637USA Ministry of Education Key Laboratory of Diagnostic Medicine and the Affiliated Hospitals of Chongqing Medical UniversityChongqing 400016China Department of Laboratory Medicine and Clinical Diagnostics The Affiliated Yantai HospitalBinzhou Medical UniversityYantai 264100China Department of Immunology and Microbiology Beijing University of Chinese MedicineBeijing 100029China

With rapid advances in understanding molecular pathogenesis of human diseases in the era of genome sciences and systems biology,it is anticipated that increasing numbers of therapeutic genes or targets will become available for targeted therapies.Despite numerous setbacks,efficacious gene and/or cell-based therapies still hold the great promise to revolutionize the clinical management of human diseases.It is wildly recognized that poor gene delivery is the limiting factor for most in vivo gene therapies.There has been a long-lasting interest in using viral vectors,especially adenoviral vectors,to deliver therapeutic genes for the past two decades.Among all currently available viral vectors,adenovirus is the most efficient gene delivery system in a broad range of cell and tissue types.The applications of adenoviral vectors in gene delivery have greatly increased in number and efficiency since their initial development.In fact,among over 2000 gene therapy clinical trials approved worldwide since 1989,a significant portion of the trials have utilized adenoviral vectors.This review aims to provide a comprehensive overview on the characteristics of adenoviral vectors,including adenoviral biology,approaches to engineering adenoviral vectors,and their applications in clinical and preclinical studies with an emphasis in the areas of cancer treatment,vaccination and regenerative medicine.Current challenges and future directions regarding the use of adenoviral vectors are also discussed.It is expected that the continued improvements in adenoviral vectors should provide great opportunities for cell and gene therapies to live up to its enormous potential in personalized medicine.

关键词： adenovirus Adenoviral vector Cell therapy Gene transfer Gene therapy Oncolytic virus Regenerative medicine Vaccine development

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adenovirus-mediated NDRG2 inhibits the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro

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Asian Pacific Journal of Tropical Medicine 2014年第11期7卷 873-878页

作者： Sheng Qiang Zhen-Fang Du Min Huang Zhangjiagang Hospital of Traditional Chinese Medicine

Objective:To investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of human renal cell carcinoma cell line OS-RC-2 in vitro.Methods:NDRG2 was harvested by RT-PCR,confirmed by DNA sequencing,and then cloned into the eukaryotic expression vector pIRES2-EGFP,which encodes green fluorescent protein(GFP),to construct p1RES2-EGFP-NDRG2 plasmid.OS-RC-2 cells with NDRG2 negative expression were transfected with p1RES2-EGFP-NDRG2 plasmid.The growth of transfected OS-RC-2 cells was observed under light and fluorescence microscopes.After colony-forming cell assays,cell proliferation detection and MTT assays,the growth curves of cells in each group were plotted to investigate the inhibitory effects of adenovirus-mediated NDRG2 on the proliferation of OS-RC-2 cells.Cell cycle was determined by flow cytometry.Confocal laser scanning microscopy showed that NDRG2 protein was specifically located on subcellular organelle.Results:A eukaryotic expression vector p1RES2-EGFP-NDRG2 was successfully constructed.After NDRG2 transfection,the growth of OS-RC-2 cells was inhibited.Flow cytometry showed that cells were arrested in S phase but the peak of cell apoptosis was not present,and confocal laser scanning microscopy showed that NDRG2 protein was located in mitochondrion.Conclusions:NDRG2 can significantly inhibit the proliferation of OS-RC-2 cells in vitro and its protein is specifically expressed in the mitochondrion.

关键词： NDRC2 OS-RC-2 adenovirus Proliferation:Inhibition

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Protective effect of thodioloside and bone marrow mesenchymal stem cells infected with HIF-1-expressing adenovirus on acute spinal cord injury

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Neural Regeneration Research 2020年第4期15卷 690-696页

作者： Xiao-Qin Ha Bo Yang Huai-Jing Hou Xiao-Ling Cai Wan-Yuan Xiong Xu-Pan Wei Lanzhou University Second Hospital LanzhouGansu ProvinceChina Department of Clinical Laboratory Lanzhou General Hospital of Lanzhou Military Area CommandLanzhouGansu ProvinceChina School of Clinical Medicine Gansu University of Traditional Chinese MedicineLanzhouGansu ProvinceChina School of Life Science and Engineering Lanzhou University of TechnologyLanzhouGansu ProvinceChina

Rhodioloside has been shown to protect cells from hypoxia injury,and bone marrow mesenchymal stem cells have a good effect on tissue repair.To study the effects of rhodioloside and bone marrow mesenchymal stem cells on spinal cord injury,a rat model of spinal cord injury was established using the Infinite Horizons method.After establishing the model,the rats were randomly divided into five groups.Rats in the control group were intragastrically injected with phosphate buffered saline(PBS)(5μL).PBS was injected at 6 equidistant points around 5 mm from the injury site and at a depth of 5 mm.Rats in the rhodioloside group were intragastrically injected with rhodioloside(5 g/kg)and intramuscularly injected with PBS.Rats in the mesenchymal stem cell(MSC)group were intramuscularly injected with PBS and intramuscularly with MSCs(8×10^6/mL in a 50-μL cell suspension).Rats in the Ad-HIF-MSC group were intragastrically injected with PBS and intramuscularly injected with HIF-1 adenovirus-infected MSCs.Rats in the rhodioloside+Ad-HIF-MSC group were intramuscularly injected with MSCs infected with the HIF-1 adenovirus and intragastrically injected with rhodioloside.One week after treatment,exercise recovery was evaluated with a modified combined behavioral score scale.Hematoxylin-eosin staining and Pischingert’s methylene blue staining were used to detect any histological or pathological changes in spinal cord tissue.Levels of adenovirus IX and Sry mRNA were detected by real-time quantitative polymerase chain reaction and used to determine the number of adenovirus and mesenchymal stem cells that were transfected into the spinal cord.Immunohistochemical staining was applied to detect HIF-1 protein levels in the spinal cord.The results showed that:(1)compared with the other groups,the rhodioloside+Ad-HIF-MSC group exhibited the highest combined behavioral score(P<0.05),the most recovered tissue,and the greatest number of neurons,as indicated by Pischingert’s methylene blue staining.(2)Compared with the PBS group,HIF-1 protein ex

关键词： acute spinal cord injury adenovirus adenovirus gene IX bone marrow mesenchymal stem cells combined behavioral score scale HIF-1α nerve regeneration nerve repair Rhodiola rosea Sry

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adenovirus-mediated LIGHT gene modification in murine B-cell lymphoma elicits a potent antitumor effect

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Cellular & Molecular Immunology 2010年第4期7卷 296-305页

作者： Guili Hu Yang Liu Hong Li Dekuang Zhao Liuqing Yang Jiangen Shen Xuejun Hong Xuetao Cao Qingqing Wang Institute of Immunology Zhejiang UniversityHangzhouChina Molecular and Cellular Biology Group Laboratory of Respiratory BiologyNIEHS/NIHResearch Triangle ParkNCUSA National Key Laboratory of Medical Immunology&Institute of Immunology Second Military Medical UniversityShanghaiChina

Here,we investigated the antitumor effect of adenovirus-mediated gene transfer of LIGHT,the tumor-necrosis factor(TNF)superfamily member also known as TNFSF14,in the murine A20 B-cell lymphoma.LIGHT gene modification resulted in upregulated expression of Fas and the accessory molecule-intercellular adhesion molecule-1(ICAM-1)on A20 cells and led to enhanced A20 cell apoptosis.LIGHT-modified A20 cells effectively stimulated the proliferation of T lymphocytes and interferon(IFN)-c production in vitro.Immunization of BALB/c mice with a LIGHT-modified A20 cell vaccine efficiently elicited protective immunity against challenge with the parental tumor cell line.adenovirus-mediated gene transfer of LIGHT by intratumoral injection exerted a very potent antitumor effect against pre-existing A20 cell lymphoma in BALB/c mice.This adenovirus-mediated LIGHT therapy induced substantial splenic natural killer(NK)and cytotoxic T lymphocyte(CTL)activity,enhanced tumor infiltration by inflammatory cells and increased chemokine expression of CC chemokine ligand 21(CCL21),IFN-inducible protein-10(IP-10)and monokine induced by IFN-c(Mig)from tumor tissues.Thus,adenovirus-mediated LIGHT therapy might have potential utility for the prevention and treatment of B-cell lymphoma.

关键词： adenovirus B lymphoma gene transfer LIGHT/TNFSF14

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Trained innate immunity and human adenovirus infection as a novel mechanism linked to the severe acute hepatitis of unknown origin in children during the COVID‐19 pandemic

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Portal Hypertension & Cirrhosis 2022年第2期1卷 145-148页

作者： Lingyao Du Jiayi Wang Lang Bai Hong Tang Center of Infectious Diseases West China Hospital of Sichuan UniversityChengduSichuanChina

1|OUTBREAK OF SEVERE ACUTE HEPATITIS OF UNKNOWN ORIGIN IN CHILDREN Severe acute hepatitis of unknown origin in children was first reported by the United Kingdom to the World Health Organization on April 5,2022.Subsequently,650 probable cases were reported worldwide by May 26,2022,1 and four patients were analyzed after reviewing the clinical records of a children's hospital in Alabama since October 1,2021.2 Although most patients recovered,the disease progressed to acute liver failure in several patients,who further required liver transplantation;thus,additional research is required to determine the disease pathogenesis.

关键词： adenovirus child innate immunity severe acute hepatitis unknown etiology

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adenovirus-mediated short hairpin RNA interference against p75 neurotrophin receptor in pheochromocytoma cells

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Neural Regeneration Research 2011年第7期6卷 517-522页

作者： Dongxu Feng Haopeng Li Siyue Xu YU Liu Xiaofei Hou Second Department of Orthopaedics Second Affiliated Hospital College of Medicine Xi'an Jiaotong University Xi'an 710004 ShaanxiProvince China First Affiliated Hospital Baotou Medical College Baotao 014010 Inner Mongolia Autonomous Region China

Previous studies have confirmed that motor neuron apoptosis in the anterior horn of the lumbosacral spinal cord is positively correlated with p75 neurotrophin receptor （p75NTR） expression in rat models of cauda equina syndrome. This study used adenovirus to carry a short hairpin RNA （shRNA） for p75NTR gene silencing, to reduce p75NTR expression in the damaged phase and to decrease motor neuron apoptosis. Three p75 siRNA template oligonucleotide segments （shRNA） were designed, and cloned into the 1.0 CMV shuttle vector. HEK293 cells were cotransfected with shuttle vector （carrying shRNA） and an adenovirus vector framework expressing enhanced green fluorescent protein. Thus, this study successfully obtained adenovirus carrying p75shRNA. The obtained viruses were named Ad.shRNA1, Ad.shRNA2, and Ad.shRNA3. The recombinant adenoviruses were separately used to infect cultured pheochromocytoma cells （PC12）. Forty-eight hours later, p75NTR mRNA and total protein were analyzed from the PC12 cells. Compared with the negative controls, RNA interference rates were separately 98.49 ± 0.68%, 95.08 ± 1.79% and 96.60 ± 1.14% at the mRNA level, and 72.89 ± 2.17%, 58.83 ± 1.15% and 59.88 ± 0.44% at the protein level in the Ad.shRNA1, Ad.shRNA2, and Ad.shRNA3 groups, respectively. Thus, recombinant adenovirus shRNA-mediated gene silencing successfully suppressed p75NTR expression.

关键词： p75 neurotrophin receptor RNA interference adenovirus rat pheochromocytoma cells human embryonic kidney 293 cells apoptosis cauda equina syndrome

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Oncolytic adenovirus targeting LASP-1 inhibited renal cell cancerprogression

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BIOCELL 2020年第4期44卷 639-647页

作者： FANGHAO SUN WENCHAO ZHAO LIANSHENG ZHANG HONGGUI MA JIAN ZHOU YOUGAN CHEN JIANQUAN HOU Department of Urology The First Affiliated Hospital of Soochow UniversitySuzhou215006China Department of Urology Soochow University Affiliated Wuxi Ninth HospitalWuxi214000China

Recent studies suggested that LIM and SH3 protein 1(LASP-1)is a promising therapeutic target for renal cell cancer(RCC).This study aimed to explore the role of LASP-1 in RCC.For this purpose,LASP-1 expression in RCC tissues was analyzed by immunohistochemistry and Western blot analysis.Cell proliferation,migration,invasion,and gene expression were detected by CCK-8 assay,Transwell assay,and Western blot analysis.The results showed that LASP-1 was highly expressed in RCC,and its expression level,t was positively correlated with lymph node metastasis and tumor,nodes,and metastases(TNM)stage.The knockdown of LASP-1 expression significantly inhibited the proliferation of RCC cells,increased the apoptosis rate,and inhibited RCC cell invasion and migration by inhibiting epithelial–mesenchymal transition.We conclude that LASP-1 promotes RCC progression and metastasis and is a promising therapeutic target for RCC.

关键词： LASP-1 Oncolytic adenovirus Renal cell cancer

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adenovirus-mediated expression of both antisense ODC and AdoMetDC inhibited colorectal cancer cell growth in vitro

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Acta Pharmacologica Sinica 2006年第3期27卷 353-359页

作者： Bing ZHANG~2 Xian-xi LIU~(2,5) Yan ZHANG~2 Chun-ying JIANG~3 Qing-shan TENG~4 Hai-yan HU~2 Wei WANG~2 Lei GONG~2 ~2Institute of Biochemistry and Molecular Biology,School of Medicine.Shandong University,Jinan 250012,China ~3Department of Coloproctology,Affiliated Hospital of Shandong University of Traditional Chinese Medicine,Jinan 250014,China,~4Department of Neurosurgery.Cleveland Clinic Foundation,Cleveland,OH 44195,USA InstituteofBiochemistryandMolecularBiology SchoolofMedicineShandongUniversityJinan250012China DepartmentofColo-proctology AffiliatedHospitalofShandongUniversityofTraditionalChineseMedicineJinan250014China

Aim:To construct a recombinant adenovirus that can simultaneously expressboth antisense ornithine decarboxylase(ODC)and adenosylmethionine decar-boxylase(AdoMetDC)and detect its inhibitory effect on the intracellular polyaminepool and colorectal cancer cell growth.Methods:A 205-bp cDNA of AdoMetDCwas reverse-inserted into recombinant pAdTrack-ODCas vectors and recombinedwith pAdEasy-1 vectors in AdEasy-1 cells.Positive clones were selected andtransfected into the packaging cell HEK293 after they were linearized by PacI.Green fluorescent protein expression was used to monitor the process of adenovi-rus packaging.The ODC and AdoMetDC protein levels were identified by west-ern blotting,and intracellular polyamine content was detected by reverse-phasehigh performance liquid chromatography.A viable cell count was used to deter-mine the growth of HT-29 cells with or without exogenous polyamine.Results:Sequencing confirmed that AdoMetDC cDNA was successfully ligated into thepAdTrack-ODCas vector.GFP expression in 293 cells during virus packing andamplification was observed by fluorescence microscopy.Western blotting dem-onstrated that both ODC and AdoMetDC were downregulated by Ad-ODC-AdoMetDCas,and consequently 3 kinds of polyamine(putrescine,spermidineand spermine)were reduced to very low levels.HT-29 cell growth was signifi-cantly inhibited as compared with control conditions,and growth arrest was notreversed by exogenous putrescine.Conclusion:The successfully constructedrecombinant adenovirus,Ad-ODC-AdoMetDCas,blocked polyamine synthesisand has therapeutic potential for treating colorectal cancer in vitro.

关键词： polyamine ornithine decarboxylase adenosylmethionine decarboxylase adenovirus colorectal cancer

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adenovirus-mediated HSV-TK Gene Therapy Using hTERT Promoter in CNE Cells in vitro

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Chemical Research in Chinese Universities 2009年第1期25卷 60-63页

作者： ZHANG Yu YU Xiang-hui ZHA Xiao KONG Wei Vaccine Research Center College of Life Science Jilin University Changchun 130012 P. R. China Department of Radiotherapy Department of Molecular Pharmacology Sichuan Tumor Hospital Chengdu 610041 P. R. China

Human telomerase reverse transcriptase（hTERT） activity was detected in human nasopharyngeal carcinoma celI（CNE） but not in human normal lung fibroblas t（CCD-11Lu）. Recombinant adenoviruses Ad-CMV-TK-enh and Ad-hTERT-TK-enh were constructed and infected into normal fibroblasts and nasopharyngeal carcinoma cells. Ad-CMV-TK-enh with 100 μmol/L of ganciclovir（GCV） caused 87% of CCD-11Lu cells death and 91% of CNE cells death, Ad-hTERT-TK-enh with 100 μmol/L of GCV caused 24% of CCD-11Lu cells death and 79% of CNE cells death. These results indicate that the Ad-hTERT-TK-enh with GCV may be a useful method in suppressing tumor growth in targeted nasopharyngeal carcinoma gene therapy.

关键词： hTERT HSV-TK adenovirus Nasopharyngeal carcinoma

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ADV36 adipogenic adenovirus in human liver disease

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World Journal of Gastroenterology 2014年第40期20卷 14706-14716页

作者： Francesca M Trovato Daniela Catalano Adriana Garozzo G Fabio Martines Clara Pirri Guglielmo M Trovato Department of Medical anPediatric Sciences Internal Medicine University of Catania95100 Catania Italy Department of Biomedical Science University of Catania 95100 Catania Italy

obesity and liver steatosis are usually described as related diseases.obesity is regarded as exclusive consequence of an imbalance between food intake and physical exercise,modulated by endocrine and genetic factors.Non-alcoholic fatty liver disease(NAFLD),is a condition whose natural history is related to,but not completely explained by over-nutrition,obesity and insulin resistance.There is evidence that environmental infections,and notably adipogenic adenoviruses(ADV)infections in humans,are associated not only with obesity,which is sufficiently established,but also with allied conditions,such as fatty liver.In order to elucidate the role,if any,of previous ADV36 infection in humans,we investigated association of ADV36-ADV37 seropositivity with obesity and fatty liver in humans.Moreover,the possibility that lifestyle-nutritional intervention in patients with NAFLD and different ADV36 seropositive status,achieves different clinical outcomes on ultrasound bright liver imaging,insulin resistance and obesity was challenged.ADV36 seropositive patientshave a more consistent decrease in insulin resistance,fatty liver severity and body weight in comparison with ADV36 seronegative patients,indicating a greater responsiveness to nutritional intervention.These effects were not dependent on a greater pre-interventional body weight and older age.These results imply that no obvious disadvantage-and,seemingly,that some benefit-is linked to ADV36 seropositivity,at least in NAFLD.ADV36 previous infection can boost weight loss and recovery of insulin sensitivity under interventional treatment.

关键词： Non-alcoholic fatty liver disease Human adenovirus

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