Preserving Genome Integrity In AAV-Mediated Gene Therapy
在 AAV 介导的基因治疗中保持基因组完整性
基本信息
- 批准号:10558679
- 负责人:
- 金额:$ 64.52万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2022
- 资助国家:美国
- 起止时间:2022-02-01 至 2026-01-31
- 项目状态:未结题
- 来源:
- 关键词:Animal ModelAnimalsAttentionBase SequenceBiologyBlood Coagulation DisordersCD8-Positive T-LymphocytesCanis familiarisCapsidCellsClonal ExpansionDNADNA IntegrationDNA RepairDNA Repair PathwayDNA StructureDataDependovirusDoseEffectivenessEventF8 geneFactor VIIIFatal OutcomeFrequenciesGene DeliveryGene ModifiedGene RearrangementGene therapy trialGenesGenetic DiseasesGenomeGenomicsGoalsHemophilia AHepatotoxicityHolliday Junction ResolvasesHumanImmune responseInsertional MutagenesisIntegration Host FactorsInverted Repeat SequencesInverted Terminal RepeatLinkLiteratureLiverLongitudinal StudiesMalignant NeoplasmsMediatingMethodsMicroRNAsModelingNatureNonhomologous DNA End JoiningOutcomeOutputPathway interactionsProductionProteinsReactionReportingRiskRisk ReductionSafetySamplingSequence AnalysisSmall Interfering RNAStructureTherapeuticTimeTissuesToxic effectTransgenesTreatment EfficacyValidationViralViral GenomeWorkadeno-associated viral vectorcell growthclinical developmentdeep sequencingdesigndog genomeexperiencegene correctiongene productgene therapygenome integritygenotoxicityhomologous recombinationimprovednext generation sequencingnovelnovel strategiesnucleaseparticlepreservationpreventrepairedsmall hairpin RNAsmall moleculesmall molecule inhibitortherapeutic genetransduction efficiencytransgene expressionvectorvector genomeviral DNA
项目摘要
ABSTRACT
Adeno-associated virus (AAV) vectors are in clinical development for delivery of genes to treat multiple genetic
diseases including hemophilia. While progress has been made to optimize gene delivery, in some studies the
required AAV vector doses were high, leading to toxicity and even fatal outcomes in one study. These findings
highlight the need for novel approaches to reduce the AAV vector dose to minimize liver toxicity, anti-AAV
immune responses, and genotoxicity. Our recent studies and work from others have identified an
underappreciated limitation to efficient gene correction with AAV vectors. In a long term study of AAV gene
delivery of FVIII in hemophilia A dogs, we found that most of the AAV vector genomes were highly rearranged
in transduced liver tissues. These rearrangements typically disrupted the transgene, and so would compromise
expression of the transgene product—unexpectedly, our data indicated that most of the AAV vector genomes
present did not produce functional protein after transduction. These rearranged AAV genomes were present in
integrated forms but also in AAV concatemers that may be episomal forms. It is unclear whether these
rearrangements occurred during vector production or after transduction of the target cells, though data is
accumulating that at least some of the rearrangements originate in vector producer cells. Our hemophilia A dog
study also identified integration events in the canine genome within genes linked to cell growth and cancer that
were associated with clonal expansions. Validation of integrated AAV DNA in these expanded clones by
sequence analysis showed that in all cases integrated vectors were highly rearranged, with only one of five
encoding an intact transgene. An extensive literature documents interactions of AAV with host DNA repair
pathways in both vector producer and target cells, though the influence of host factors in AAV DNA
rearrangements is mostly unstudied. We hypothesize that modulation of host cell pathways can suppress AAV
DNA rearrangements, thereby allowing improved transgene expression per vector DNA copy. In this proposal,
we will 1) implement a deep sequencing method to quantify rearrangement frequency in a statistically rigorous
fashion, 2) identify cellular pathways that can be modulated with small molecules, siRNAs, or microRNAs that
suppress vector rearrangements, and 3) devise novel delivery strategies that support efficient pathway
modulation, suppress vector rearrangement, and boost transgene output per vector copy. These methods will
be assessed during AAV vector production (Specific Aim 1) and after AAV delivery in the transduced target cells
(Specific Aim 2). Our deliverables at the end of the project will be a greatly enhanced understanding of the
interaction of AAV with host cell DNA handling pathways, and methods for modulating these pathways to allow
safe and effective gene delivery at lower vector doses.
摘要
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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Frederic D Bushman其他文献
The enigmatic roles of emAnelloviridae/em and emRedondoviridae/em in humans
人类中的 emAnelloviridae/em 和 emRedondoviridae/em 的神秘作用
- DOI:
10.1016/j.coviro.2022.101248 - 发表时间:
2022-08-01 - 期刊:
- 影响因子:5.100
- 作者:
Louis J Taylor;Emma L Keeler;Frederic D Bushman;Ronald G Collman - 通讯作者:
Ronald G Collman
HTLV-1 clonality during chronic infection and BLV clonality during primary infection
- DOI:
10.1186/1742-4690-8-s1-a185 - 发表时间:
2011-06-06 - 期刊:
- 影响因子:3.900
- 作者:
Nicolas A Gillet;Carol Hlela;Tine Verdonck;Eduardo Gotuzzo;Daniel Clark;Sabrina Rodriguez;Nirav Malani;Anat Melamed;Niall Gormley;Richard Carter;David Bentley;Charles Berry;Frederic D Bushman;Graham P Taylor;Luc Willems;Charles R M Bangham - 通讯作者:
Charles R M Bangham
Bromodomain and extra-terminal (BET) proteins target Moloney murine leukemia virus integration to transcription start sites
- DOI:
10.1186/1742-4690-10-s1-o20 - 发表时间:
2013-09-19 - 期刊:
- 影响因子:3.900
- 作者:
Jan De Riick;Christine de Kogel;Jonas Demeulemeester;Sofie Vets;Nirav Malani;Frederic D Bushman;Katrien Busschots;Steven Husson;Rik Gijsbers;Zeger Debyser - 通讯作者:
Zeger Debyser
Frederic D Bushman的其他文献
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{{ truncateString('Frederic D Bushman', 18)}}的其他基金
mVACS--mRNA Vaccines for C. difficile Suppression
mVACS--用于抑制艰难梭菌的 mRNA 疫苗
- 批准号:
10625573 - 财政年份:2023
- 资助金额:
$ 64.52万 - 项目类别:
Preserving Genome Integrity In AAV-Mediated Gene Therapy
在 AAV 介导的基因治疗中保持基因组完整性
- 批准号:
10338480 - 财政年份:2022
- 资助金额:
$ 64.52万 - 项目类别:
Linking insertional mutagenesis and cell function to improve CAR T cell therapy
将插入突变与细胞功能联系起来以改善 CAR T 细胞疗法
- 批准号:
10398224 - 财政年份:2019
- 资助金额:
$ 64.52万 - 项目类别:
Linking insertional mutagenesis and cell function to improve CAR T cell therapy
将插入突变与细胞功能联系起来以改善 CAR T 细胞疗法
- 批准号:
10158019 - 财政年份:2019
- 资助金额:
$ 64.52万 - 项目类别:
Linking insertional mutagenesis and cell function to improve CAR T cell therapy
将插入突变与细胞功能联系起来以改善 CAR T 细胞疗法
- 批准号:
10640072 - 财政年份:2019
- 资助金额:
$ 64.52万 - 项目类别:
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