New horizons in translational medicine最新文献

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Autophagic flux without a block differentiates varicella from herpes simplex virus infection 无阻断的自噬通量可区分水痘与单纯疱疹病毒感染
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.066
Charles Grose
{"title":"Autophagic flux without a block differentiates varicella from herpes simplex virus infection","authors":"Charles Grose","doi":"10.1016/j.nhtm.2015.07.066","DOIUrl":"10.1016/j.nhtm.2015.07.066","url":null,"abstract":"<div><p><span><span>Varicella-zoster virus (VZV) is a </span>herpesvirus<span> that causes a characteristic vesicular exanthem in humans with primary infection (varicella) or reactivation (zoster). We have previously observed that vesicular cells are filled with autophagosomes that are easily detectable by </span></span>confocal microscopy<span><span> after immunolabeling for the LC3 protein. Through a </span>3D imaging software program called Imaris we have quantitated autophagosomes as greater than 100 per cell. Similarly, we have assessed autophagy in VZV-infected monolayers after inoculation by the traditional method with infected cells at a ratio of one infected to 8 uninfected cells. Again, autophagosomes are easily detected, but their count is lower than that observed in human skin cells. As an additional control, we enumerated the autophagosomes in the Severe Combined Immuno-Deficient (SCID) Mouse model of VZV infection. In this model, human skin is inserted under the skin of the mouse and subsequently inoculated with VZV-infected cells. Again, autophagy was abundant in the VZV-infected skin and minimal in the mock-infected skin sample. Subsequently, we investigated autophagy following infection with sonically prepared cell free virus in cultured cells. After cell free virus inoculation, autophagy was detected in a majority of infected cells at all time points, but was less than that seen after an infected-cell inoculum. Finally, we investigated VZV-induced autophagic flux by two different methods (radiolabeling proteins and a dual-colored LC3 plasmid); both showed no evidence of a block in autophagy. Overall, therefore, autophagy within a VZV-infected cell was remarkably different from autophagy within an HSV-infected cell, whose genome contains two modifiers of autophagy, ICP34.5 and US11, not present in VZV.</span></p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Page 133"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.066","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80566964","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Animation of VZV DNA VZV DNA动画
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.064
Randall. J. Cohrs , J. Rovnak
{"title":"Animation of VZV DNA","authors":"Randall. J. Cohrs ,&nbsp;J. Rovnak","doi":"10.1016/j.nhtm.2015.07.064","DOIUrl":"10.1016/j.nhtm.2015.07.064","url":null,"abstract":"<div><p>Varicella zoster virus<span> (VZV) is a ubiquitous neurotropic alphaherpesvirus that typically causes childhood varicella (chickenpox) on primary infection and zoster<span><span> (shingles) after reactivation. During latency most of the ~70 virus genes are transcriptionally silent; however, analysis of latent VZV gene transcription in its natural setting requires analysis of human </span>ganglia<span> removed at autopsy. Recognizing the problems associated with such samples, we have observed that as the post-mortem time interval increases, so do the number of VZV genes transcribed. Based on our data and recent similar findings concerning reactivation of HSV-1, we propose an interesting testable model to describe epigenetic control of neurotropic alphaherpesvirus gene transcription during latency and early reactivation.</span></span></span></p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Page 133"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.064","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88629421","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Predicting Prion Propensity of Human Proteins 预测人类蛋白质的朊病毒倾向
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.042
Cascarina S, Ross E.
{"title":"Predicting Prion Propensity of Human Proteins","authors":"Cascarina S,&nbsp;Ross E.","doi":"10.1016/j.nhtm.2015.07.042","DOIUrl":"10.1016/j.nhtm.2015.07.042","url":null,"abstract":"<div><p>In humans only a single prion-forming protein named PrP<sup>c</sup> (for “<u>c</u>ellular <u>pr</u>ion <u>p</u><span>rotein”) is currently known, yet many more neurodegenerative disorders<span> involve aberrant protein aggregation. The classical model for these diseases has involved cell-autonomous aggregation, assuming that aggregation occurs independently in each cell within a diseased patient. However, more recent models have proposed a non-cell-autonomous progression of disease in which aggregates formed in one cell may be transmitted to neighboring cells. These aggregate seeds then cause aggregation of the soluble protein in the “infected” cells, similar to the prion diseases<span>. Within the past few years, a number of proteins that exhibit prion-like aggregation and spread to neighboring tissues have been discovered in patients<span> with Amyotrophic Lateral Sclerosis (ALS). Although ALS has been studied for a number of decades, these proteins were only recently linked to ALS by chance. This demonstrates a clear need for an accurate method to systematically identify additional proteins that may play a pathological role in neurodegenerative disorders. Taking advantage of the compositional similarity of these proteins to the known yeast prions, I plan to use the prion prediction methodology that our lab has pioneered to develop an entirely new algorithm specifically suited for this class of neuronal proteins.</span></span></span></span></p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Page 127"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.042","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88977341","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Transcriptome markers of viral persistence in naturally-infected Andes Hantavirus (Bunyaviridae) seropositive rice rats 自然感染安第斯汉坦病毒(布尼亚病毒科)血清阳性水稻大鼠中病毒持久性的转录组标记物
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.063
Corey L. Campbell , Fernando Torres-Perez , Mariana Acuna-Retamar , Tony Schountz
{"title":"Transcriptome markers of viral persistence in naturally-infected Andes Hantavirus (Bunyaviridae) seropositive rice rats","authors":"Corey L. Campbell ,&nbsp;Fernando Torres-Perez ,&nbsp;Mariana Acuna-Retamar ,&nbsp;Tony Schountz","doi":"10.1016/j.nhtm.2015.07.063","DOIUrl":"10.1016/j.nhtm.2015.07.063","url":null,"abstract":"<div><p>The long-tailed pygmy rice rat (<em>Oligoryzomys longicaudatus</em><span>) is the reservoir host of Andes (ANDV) and Oran hantaviruses (</span><span><em>Bunyaviridae</em></span><span><span>). To examine transcriptome features of persistently infected rice rats, spleens from ANDV sero-positive wild-caught rice rats were assessed. RNA-seq analysis, de novo reference-independent assembly and stringent orthology assignments produced 17,756 unique coding and non-coding RNAs. Differential expression analysis of persistently-infected seropositive rice rat spleens revealed 18 differentially expressed transcripts from 16 unique genes. A three-pronged effect on the immune response were observed in 1) suppression of the JAK-STAT pathway at </span>Stat5b and Ccnot1, as well as 2) a bias toward a TH2 response in the enrichment of caspase-1 and 3) stimulation of RIG-I pathway factors Ppp1cc and MFF. Two of these differentially expressed transcripts, caspase-1 and STAT5b, code for proteins expected to stimulate T helper follicular (T</span><sub>FH</sub>) cell development, a phenomenon that has also been described for hantavirus-infected <em>P. maniculatus</em>. Differential expression of a single seropositive rice rat with a higher viral load revealed a robust response of 243 differentially expressed transcripts, suggesting an acute infection. Together, these data help define the fundamental features of the immune response in a hantavirus reservoir host.</p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Pages 132-133"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.063","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75525856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Further Characterization of Rio Grande Virus and Potential for Serological Cross Reactivity with other Phleboviruses 格兰德河病毒的进一步鉴定及其与其他静脉病毒的血清学交叉反应潜力
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.059
Szymczak M , Reeves W , Miller M
{"title":"Further Characterization of Rio Grande Virus and Potential for Serological Cross Reactivity with other Phleboviruses","authors":"Szymczak M ,&nbsp;Reeves W ,&nbsp;Miller M","doi":"10.1016/j.nhtm.2015.07.059","DOIUrl":"10.1016/j.nhtm.2015.07.059","url":null,"abstract":"<div><p>Members of the genus <em>Phlebovirus</em> (family <span><em>Bunyaviridae</em></span><span><span>) are new and emerging disease pathogens<span><span> of humans and animals. Newly identified viruses include Heartland virus (HRTV), Lone Star virus in the USA, and </span>Severe Fever with Thrombocytopenia Syndrome virus in Asia. Assays to support surveillance, </span></span>epidemiologic studies, and diagnosis of these viruses may also detect related viruses within the genus, confounding interpretation. Rio Grande virus (RGV) was isolated in 1973 from southern plains woodrats (</span><em>Neotoma micropus</em>) in the United States and has been preliminarily identified as a phlebovirus transmitted by the sand fly <span><em>Lutzomyia</em><em> anthophora</em></span><span><span><span>. RGV is not known to cause disease in humans, but it could be detected by assays designed for HRTV or other phleboviruses. The goal of this study was to determine antigenic cross-reaction between RGV and other phleboviruses. A commercially available ELISA based sand fly fever antigen detection kit was tested for the ability to detect RGV and other New and Old-World phleboviruses, including attenuated Rift valley fever virus (RVFV) strain MP12, </span>Punta Toro virus (PTV), </span>Toscana virus<span><span><span><span>, Aguacate virus, Anhanga virus, Arumowot virus, and Chagres virus. Immunocytochemistry and </span>Western blotting were used to detect cross reactions between RGV, MP12, and PTV using rabbit anti-RVFV nucleocapsid protein and </span>glycoproteins GC and GN, mouse monoclonal anti-PTV, and sheep polyclonal anti-MP12. The ELISA test detected cross reactivity for all phleboviruses excluding RGV, but Western blotting detected the presumed </span>RGV nucleocapsid protein (N) using rabbit anti-RVFV-N serum, RGV-infected cells were also identified when labeled with this antibody. Our findings demonstrate assay specific antigenic cross reactivity between these phleboviruses, thus further characterization of the molecular targets of the cross-reaction is required for proper interpretation of serological assays.</span></span></p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Pages 131-132"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.059","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84243620","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
The Intrathecal Antibody Response in Multiple Sclerosis Brain Does Not React Against Measles Virus 多发性硬化症脑鞘内抗体反应对麻疹病毒没有反应
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.060
Deandra L Walker, Mark P Burgoon
{"title":"The Intrathecal Antibody Response in Multiple Sclerosis Brain Does Not React Against Measles Virus","authors":"Deandra L Walker,&nbsp;Mark P Burgoon","doi":"10.1016/j.nhtm.2015.07.060","DOIUrl":"10.1016/j.nhtm.2015.07.060","url":null,"abstract":"<div><p><span><span><span><span>Multiple sclerosis<span> (MS) is a chronic inflammatory demyelinating disease of the </span></span>central nervous system (CNS) and is the most common disabling </span>neurological disease of young adults. Although the cause of MS is unknown, genetic and epidemiological studies indicate that MS may be triggered by an environmental agent. The presence of intrathecally produced antibodies, which produce oligoclonal Ig bands in the CNS of MS patients, provides tools for investigating the target of the inflammatory response. In most of the CNS conditions with </span>oligoclonal bands<span> the target is known and the antibody is directed against an infectious, causative agent. For example, in subacute sclerosing panencephalitis, a </span></span>measles virus<span><span> (MV) infection of the brain, the oligoclonal bands and intrathecal antibodies are primarily directed against MV. In earlier studies of MS, we demonstrated that the intrathecal antibody response in MS brain does not react to </span>varicella zoster<span> or Epstein-Barr virus. The current study investigates the reactivity of the intrathecal antibody response in MS brain to MV. We isolated individual CD38(+) plasma cells from MS brain to produce recombinant antibodies (rAbs). These rAbs likely represent oligoclonal bands and were used to immunostain MV-infected or uninfected monkey kidney (Vero) cells. Although many of the rAbs from MS brain reacted against auto-antigens in several mouse and human tissues, none of fifteen MS rAbs reacted against MV-infected cells. These results indicate that measles virus is not a disease-relevant antigen in MS.</span></span></p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Page 132"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.060","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80242078","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Ivermectin for the Control of West Nile Virus Transmission 控制西尼罗病毒传播的伊维菌素
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.043
Nguyen C, Burton T, Kuklinski W, Gray M, Foy BD
{"title":"Ivermectin for the Control of West Nile Virus Transmission","authors":"Nguyen C,&nbsp;Burton T,&nbsp;Kuklinski W,&nbsp;Gray M,&nbsp;Foy BD","doi":"10.1016/j.nhtm.2015.07.043","DOIUrl":"10.1016/j.nhtm.2015.07.043","url":null,"abstract":"<div><p><span><span>Presently there are limited options for controlling the transmission of West Nile virus (WNV), including the use of </span>larvicides and adulticides to target the mosquito vector. However, these methods are poorly-targeted, restricted to wealthy semi-urban and urban areas that are able to fund the efforts, and opposed in some communities due to toxicity concerns. This study evaluated the use of endectocide-treated bird feed to control WNV transmission by targeting the primary vector in Colorado, </span><em>Culex tarsalis.</em><span> Ivermectin susceptibility in </span><em>C. tarsalis</em> was first measured through ivermectin-spiked bloodmeals fed using membrane feeders, and the LC<sub>50</sub><span> was determined to be 49.94 ng/ml (39.71-59.93 95% CI, n=988). Chickens were then fed ivermectin-treated feed to examine its safety and palatability, and mosquitoes were blood fed directly on the chickens to assess in vivo effects. Finally, ivermectin pharmokinetics were analyzed using vein blood from chickens as well the </span><em>C. tarsalis</em> that bloodfed on the chickens. A mixture of 200<!--> <!-->mg ivermectin/kg of bird feed was determined to be a palatable and safe dose on which chickens would feed while also being effective in killing <em>C. tarsalis</em><span> in bioassays. Pharmacokinetic<span> data from the in vivo tests produced conflicting results compared to in vitro blood feeds but drug was detected in chicken blood at concentrations that may be expected to affect C. tarsalis. Dosing, safety, and bioassays are currently being conducted in doves and sparrows. Additional studies are currently determining the effect of ivermectin on mortality in WNV-infected mosquitoes, as well as if ivermectin reduces WNV replication and transmission. Our study indicates that the use of ivermectin-treated bird feed could be a novel method of controlling WNV transmission.</span></span></p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Page 127"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.043","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"75310126","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 3
Roche Research Portfolio: Trusted Performance, Efficient Workflow Solutions 罗氏研究组合:可靠的性能,高效的工作流程解决方案
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.068
Konet D
{"title":"Roche Research Portfolio: Trusted Performance, Efficient Workflow Solutions","authors":"Konet D","doi":"10.1016/j.nhtm.2015.07.068","DOIUrl":"10.1016/j.nhtm.2015.07.068","url":null,"abstract":"<div><p>Along with Roche Pharmaceuticals, Roche Diagnostics is an important part of the foundation that modern healthcare builds upon. Our broad range of innovative diagnostic tests and systems play a pivotal role in the groundbreaking area of integrated healthcare solutions and cover the early detection, targeted screening, evaluation and monitoring of disease. Roche Diagnostics is active in all market segments, from scientific research and clinical laboratory systems to patient self-monitoring.</p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Page 134"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.068","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85958665","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Longitudinal analysis of blood-borne prion infection 血源性朊病毒感染的纵向分析
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.07.045
Alan M. Elder , Davin M. Henderson , Amy V. Nalls , Anthony E. Kincaid , Edward A. Hoover , Jason C. Bartz , Candace K. Mathiason
{"title":"Longitudinal analysis of blood-borne prion infection","authors":"Alan M. Elder ,&nbsp;Davin M. Henderson ,&nbsp;Amy V. Nalls ,&nbsp;Anthony E. Kincaid ,&nbsp;Edward A. Hoover ,&nbsp;Jason C. Bartz ,&nbsp;Candace K. Mathiason","doi":"10.1016/j.nhtm.2015.07.045","DOIUrl":"10.1016/j.nhtm.2015.07.045","url":null,"abstract":"<div><p><span>Transmissible spongiform encephalopathies<span> (TSEs), or prion diseases, affecting human and animal species can be transmitted from TSE-infected individuals to naïve susceptible hosts during the long asymptomatic (years to decades) and symptomatic disease stages. The presence of infectious hematogenous prions in asymptomatic TSE-infected hosts demonstrates the highly infectious nature of blood-borne prions in hosts lacking overt clinical symptoms. It is currently unknown when and how infectious prions first enter the blood following initial exposure. We have previously shown that the whole-blood real-time quaking-induced conversion assay (wbRT-QuIC) possesses 100% specificity and &gt;92% sensitivity, making it an ideal tool to address this question. Here, we use wbRT-QuIC to analyze whole blood collected from oral, extranasal or aerosol TSE-exposed hosts for blood-borne prions. Our results demonstrate that conversion competent prions in the inoculum are capable of crossing mucosal surfaces<span> and entering the circulatory system within 30</span></span></span> <!-->min—no matter the route of exposure. Detection of the inoculum minutes post exposure is followed by a steady decline in the detection of blood-borne prions up to 3 days which is followed by a progressive increase in the detection of nascent conversion competent prions between 1 and 17 months post exposure. These data provide the first evidence for the facile transport of mucosally acquired prions into the circulatory system, providing evidence for multiple routes of inter- and intra- host prion trafficking and shedding.</p></div>","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Page 128"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.07.045","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87327988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 1
Signal regulatory protein alpha (SIRPA) and kinase domain receptor (KDR) are key expression markers in cardiac specific precursor selection from hADSCs 信号调节蛋白α (SIRPA)和激酶结构域受体(KDR)是hascs中心脏特异性前体选择的关键表达标记
New horizons in translational medicine Pub Date : 2015-05-01 DOI: 10.1016/j.nhtm.2015.02.001
Vinod Kumar Verma , Syed Sultan Beevi , Tanya Debnath , Usha Shalini , Suguna Ratnakar Kamaraju , Lakshmi Kumari Kona , Yamuna Mohanram , Lakshmi Kiran Chelluri
{"title":"Signal regulatory protein alpha (SIRPA) and kinase domain receptor (KDR) are key expression markers in cardiac specific precursor selection from hADSCs","authors":"Vinod Kumar Verma ,&nbsp;Syed Sultan Beevi ,&nbsp;Tanya Debnath ,&nbsp;Usha Shalini ,&nbsp;Suguna Ratnakar Kamaraju ,&nbsp;Lakshmi Kumari Kona ,&nbsp;Yamuna Mohanram ,&nbsp;Lakshmi Kiran Chelluri","doi":"10.1016/j.nhtm.2015.02.001","DOIUrl":"10.1016/j.nhtm.2015.02.001","url":null,"abstract":"<div><h3>Background</h3><p><span>Cardiomyocyte enrichment strategies so far have not yielded scalable cardiac specific cell type. More so, the current data is restricted to </span>embryonic stem cells<span><span> (ESCs)/induced pluripotent stem cells (iPSCs), wherein the use of viral vectors is fraught with increased risk during clinical use. Herein, we profiled time-dependent gene/protein expression patterns across the cardiac ectoderm, </span>endoderm<span>, and mesoderm<span> for isolating cardiac precursors from human adipose derived stem cells (hADSC).</span></span></span></p></div><div><h3>Methods</h3><p><span><span>Direct cardiac differentiation of hADSCs<span> was carried out with 5-azacytidine and basic fibroblast growth factor (bFGF) in a one month long culture. The cells were periodically harvested, analyzed for unique persistent markers and their inherent regulation using quantitative polymerase chain reaction (qPCR), flow cytometry, </span></span>immunoblot<span> and immunocytochemistry assays. The identified markers were </span></span>super paramagnetic iron oxide nanoparticle<span> (SPION) tagged for segregation by magnetic activated cell sorting (MACS) and further evaluated their differentiation potential and checked for the purity by flow cytometry.</span></p></div><div><h3>Results</h3><p><span>The results demonstrated pronounced up-regulation of mesodermal and mature cardiac lineage markers<span> at three weeks, while there was a down-regulation of pluripotent stem cell markers. This perhaps could be attributed to de-differentiation in maintaining the cardiac phenotype. However, signal </span></span>regulatory protein alpha (SIRPA) and kinase domain receptor (KDR) persisted all through the culture period of one month, making them the most relevant and reliable cardiac specific markers. Dual labeling of these markers to SPION for cardiomyocyte enrichment by MACS column yielded cardiomyogenic-like cells in differentiation cultures with several functional positive markers.</p></div><div><h3>Conclusions</h3><p>Thus, SIRPA and KDR together provide cues in the enhancement and up-scaling of cardiomyocyte production in the cell replacement therapy.</p></div><div><h3>Focal points</h3><p></p><ul><li><span>•</span><span><p>Benchside</p><p>Identification of specific cell phenotypic markers to identify cardiac precursors in any tissue source with minimal cell manipulation is a novel process development tool in clinical translation.</p></span></li><li><span>•</span><span><p>Bedside</p><p>A product developed in a closed system would minimize extraneous contaminants in long term cultures and development of such procedures minimizes culture failure rates from bench side.</p></span></li><li><span>•</span><span><p>Industry</p><p>This unique identification of cell-specific marker would enable a tissue-specific translational plan and immensely help in the cardiac regeneration.</p></span></li><li><span>•</span><span><p>Government</p><p>Financial inve","PeriodicalId":90660,"journal":{"name":"New horizons in translational medicine","volume":"2 4","pages":"Pages 93-101"},"PeriodicalIF":0.0,"publicationDate":"2015-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.nhtm.2015.02.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"78417284","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 4
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