Project 1: KSHV reprograms replication and metabolic activities in hypoxia
Project 1: KSHV reprograms replication and metabolic activities in hypoxia
批准号:
10714173
负责人:
ERLE S. ROBERTSON
金额:
$46.85万
依托单位国家:
美国
项目类别:
财政年份:
2023
资助国家:
美国
项目状态:
未结题
起止时间:
2023-09-01 至 2028-08-31
关键词:
AddressAnimal ModelAntigensAutomobile DrivingB-LymphocytesBiological AssayCell Culture SystemCell CycleCell Cycle ProteinsCell HypoxiaCell LineCell ProliferationCell ReprogrammingCellsChemicalsClustered Regularly Interspaced Short Palindromic RepeatsCytoprotectionDNA VirusesDNA biosynthesisDataDevelopmentDrug TargetingEndothelial CellsEnvironmentEnzyme InductionEnzymesEpigenetic ProcessEtiologyFamilyGenerationsGenesGenetic TranscriptionGenomeGoalsHIF1A geneHarvestHerpesviridaeHerpesviridae InfectionsHumanHuman Cell LineHuman Herpesvirus 8HypoxiaImmunocompromised HostIn VitroIncubatedIndividualKaposi SarcomaKnock-outLesionLinkLymphomaLymphoproliferative DisordersMalignant NeoplasmsMediatingMetabolicMetabolismModificationMolecularMulticentric Angiofollicular Lymphoid HyperplasiaNucleotidesOncogenic VirusesOncornavirusesPathogenicityPathologyPathway interactionsPhasePleural effusion disorderPopulationPost Translational Modification AnalysisPost-Translational Protein ProcessingPredispositionProductionProteinsProteomicsRegulationRoleStressSystemTetanus Helper PeptideTimeTranscriptional RegulationTranslationsViralViral AntigensViral GenomeVirusVirus LatencyVirus ReplicationWestern Blottingdaughter cellinfected B cellinsightknock-downlytic replicationmembernormoxianovelprimary effusion lymphomaprogramssmall hairpin RNAthree dimensional cell culturetranscriptometumorigenesisvirus related cancer
中文摘要
项目#1
摘要
KSHV相关癌症发生在缺氧微环境中。缺氧对病毒和宿主DNA的影响
在KSHV感染的情况下的复制知之甚少。我们以前研究过复制
KSHV在常氧细胞培养系统中的表达。我们现在提出一个深入的分析KSHV和感染的宿主细胞
在低氧环境中复制。KSHV,像疱疹病毒家族的许多成员一样,
潜伏和裂解复制周期。在细胞培养系统中,潜伏期是通过细胞培养物的同步化来维持的。
病毒基因组的复制与宿主细胞的复制是细胞周期依赖性的。所以大部分的
潜伏复制活动在每个细胞周期利用宿主机器一次。这些潜伏感染的细胞通常
通过用化学诱导剂处理诱导裂解性复制,并且需要表达许多
病毒编码基因然而,关于病毒基因组在缺氧中的复制知之甚少,并且病毒基因组在缺氧环境中的复制是不稳定的。
抗原需要调节正在进行的代谢变化所需的持久性的病毒基因组,
在受感染的细胞中复制,从而允许繁殖成新的子细胞。我们将探讨这些问题
通过一系列旨在了解细胞和病毒复制蛋白质变化的目标,
与常氧时相比,缺氧时会出现这种情况。我们将研究细胞复制的变化
细胞周期不同阶段的蛋白质及其翻译后修饰。它们在缺氧状态下的水平
以确定复制机器的基本组成部分的变化。另外我们
我会看看他们的转录,以确定这些变化是否是由于在整体转录的变化,
基因,或者如果变化是由于缺氧时HIF 1 α的转录调节。代谢的变化
还将分析与低氧中KSHV阴性细胞相比,在KSHV感染细胞中诱导的酶
以确定这些变化是否是转录重编程。我们将确定代谢基因,
如果复制基因受HIF 1 α调节,则主基因在低氧条件下对KSHV感染的影响可能是异常的。
转录调节因子。我们将确定缺氧期间发生的表观遗传变化,
确定复制基因的表达。我们还将在3D培养系统中进行这些测定,
模仿相关的病理。我们还将产生shRNA敲除或CRISPR敲除的选择的
复制基因和Tet-regulated基因,以确定这些蛋白质在KSHV复制中的作用。
缺氧微环境
14
英文摘要
Project #1
Abstract
KSHV-associated cancers occur in hypoxic microenvironments. The effects of hypoxia on viral and host DNA
replication in the context of KSHV infection are poorly understood. We have previously investigated the replication
of KSHV in normoxia cell culture systems. We now propose an in-depth analysis of KSHV and infected host cell
replication in the hypoxic environment. KSHV, like many members of the herpesvirus family undergoes both a
latent and a lytic replication cycle. In cell culture systems, latency is maintained through synchronization of the
viral genome replication with that of the host cell replication, which is cell cycle dependent. Therefore, most of the
latent replication activities utilize the host machinery once per cell cycle. These latently infected cells are typically
induced to lytic replication through treatment with chemical inducers and requires the expression of a number of
viral encoded genes. However, little is known regarding replication of the virus genome in hypoxia, and the viral
antigens needed to regulate the ongoing metabolic changes required for persistence of the viral genome through
replication in infected cells that allow for propagation into new daughter cells. We will approach these questions
through a number of aims geared towards understanding the changes in the cellular and viral replication proteins
that occur in hypoxia compared to that seen in normoxia. We will examine the changes in cellular replication
proteins at different phases of the cell cycle, and their post-translational modification. Their levels in hypoxia will
be analyzed to determine the changes in the essential components of the replication machinery. In addition, we
will look at their transcription to determine if these changes are due to changes in the overall transcription of the
genes or if the changes are due to transcription modulation by HIF1α in hypoxia. The changes in metabolic
enzymes that are induced in KSHV infected cells compared to KSHV negative cells in hypoxia will also be analyzed
to determine if the changes are transcriptionally reprogrammed. We will determine the metabolic genes that are
dysregulated on KSHV infection in hypoxia and if the replication genes are regulated by HIF1α, the master
transcription regulator in hypoxia. We will determine the epigenetic changes that occur during hypoxia that will
determine the expression of the replication genes. We will also perform these assays in 3D culture systems to
mimic the associated pathologies. We will also generate shRNA knockdown or CRISPR knockout of selected
replication genes, and Tet-regulated genes to determine the contribution of these proteins in KSHV replication in
the hypoxic microenvironment.
14
期刊论文(0)
专著(0)
科研奖励(0)
会议论文
Botswana-UPenn: Research Consortium of HPV-Related Cervical Cancer in HIV Patient
-
批准号:10834480
-
项目类别:
-
资助金额:$6.93万
-
财政年份:2023
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Transcription and Replication of Oncogenic Viruses in Hypoxia
-
批准号:10714172
-
项目类别:
-
资助金额:$269.7万
-
财政年份:2023
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Virus, Vector and Cell Culture Core
-
批准号:10714178
-
项目类别:
-
资助金额:$30.95万
-
财政年份:2023
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Administrative Core
-
批准号:10714177
-
项目类别:
-
资助金额:$11.33万
-
财政年份:2023
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Project 2: KSHV induces tumorigenesis by harnessing differentiation in hypoxia
-
批准号:10714174
-
项目类别:
-
资助金额:$45.55万
-
财政年份:2023
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Next Generation Sequencing Core
-
批准号:10714179
-
项目类别:
-
资助金额:$45.67万
-
财政年份:2023
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Tumor suppressor reprogramming by EBV through post-translational modification
-
批准号:10402055
-
项目类别:
-
资助金额:$53.94万
-
财政年份:2022
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Tumor suppressor reprogramming by EBV through post-translational modification
-
批准号:10684650
-
项目类别:
-
资助金额:$53.33万
-
财政年份:2022
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Epigenetic Regulation of KSHV Genome Replication
-
批准号:10457380
-
项目类别:
-
资助金额:$50.18万
-
财政年份:2019
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Epigenetic Regulation of KSHV Genome Replication
-
批准号:9978759
-
项目类别:
-
资助金额:$49.57万
-
财政年份:2019
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Epigenetic Regulation of KSHV Genome Replication
-
批准号:10669729
-
项目类别:
-
资助金额:$50.18万
-
财政年份:2019
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Epigenetic Regulation of KSHV Genome Replication
-
批准号:10208828
-
项目类别:
-
资助金额:$50.18万
-
财政年份:2019
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Natural History & Pathogenesis of HPV in HIV infected women with cervical cancer
-
批准号:8936659
-
项目类别:
-
资助金额:$10.55万
-
财政年份:2014
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Botswana-UPENN Research Consortium of HPV-related Cervical Cancer in HIV Patients
-
批准号:8794812
-
项目类别:
-
资助金额:$75.0万
-
财政年份:2014
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Botswana-UPENN Research Consortium of HPV-related Cervical Cancer in HIV Patients
-
批准号:9770545
-
项目类别:
-
资助金额:$95.29万
-
财政年份:2014
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Botswana-UPENN Research Consortium of HPV-related Cervical Cancer in HIV Patients
-
批准号:9128422
-
项目类别:
-
资助金额:$5.0万
-
财政年份:2014
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Botswana-UPENN Research Consortium of HPV-related Cervical Cancer in HIV Patients
-
批准号:8927590
-
项目类别:
-
资助金额:$74.78万
-
财政年份:2014
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Early Events in KSVH Infection of Primary B-cells
-
批准号:8540468
-
项目类别:
-
资助金额:$186.18万
-
财政年份:2013
-
负责人:ERLE S. ROBERTSON
-
依托单位:
Genome Persistence of KSHV
-
批准号:8467401
-
项目类别:
-
资助金额:$29.88万
-
财政年份:2013
-
负责人:ERLE S. ROBERTSON
-
依托单位:
KSHV genome modification in KS tissue
-
批准号:8926364
-
项目类别:
-
资助金额:$20.0万
-
财政年份:2013
-
负责人:ERLE S. ROBERTSON
-
依托单位:
海外基金