Base editing and prime editing for sickle cell disease
镰状细胞病的碱基编辑和引物编辑
基本信息
- 批准号:10579903
- 负责人:
- 金额:$ 77.71万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-01-01 至 2024-12-31
- 项目状态:已结题
- 来源:
- 关键词:Acute PainAdenosineAdultAffectAffinityAlanineAllelesAllogenicAmericanAmino AcidsAntisickling AgentsArchitectureAutologousBase PairingBenignBiochemicalBiological AssayBloodCD34 geneCaringCell DeathCell LineCell TherapyCell physiologyCellsCessation of lifeChromosomal RearrangementClinical ResearchCodon NucleotidesComplexDNADNA Double Strand BreakDeteriorationDevelopmentDiseaseElementsEngineeringEngraftmentEnzymesErythrocytesErythroid CellsErythroid Progenitor CellsEscherichia coliEvolutionFetal HemoglobinFrequenciesFunctional disorderFutureGene ExpressionGene SilencingGenesGeneticGenetic DiseasesGenetic EngineeringGenetic TemplateGenomeGenomic DNAGenomicsGlobinGuide RNAHLA AntigensHematological DiseaseHematopoieticHematopoietic Stem Cell TransplantationHematopoietic stem cellsHemoglobinHemoglobin F DiseaseHemolytic AnemiaHumanHypoxiaIn VitroIndividualInheritedMaintenanceMalignant - descriptorMediatingMedicalMessenger RNAMethodsMissense MutationModificationMorbidity - disease rateMultiple Organ FailureMusMutationNucleotidesOrganOutcomePainPatientsPolymersProteinsQuality of lifeRNAReagentRecombinantsRegulationResearchSafetySickle CellSickle Cell AnemiaSickle Cell TraitSickle HemoglobinSiteTechnologyTestingTherapeuticToxic effectTranscriptional Silencer ElementsTransplantationTreatment EfficacyValineVariantXenograft procedurebase editingbase editorbeta Globinchronic paincurative treatmentsexperiencegene therapygenetic approachgenetic informationgenetic manipulationgenome editingimprovedin vivoinsightmanufacturemortalitymouse modelmutantnew technologynovelnovel strategiesoff-target mutationpolymerizationprecision geneticsprematurepreventprime editingprime editorpromoterrepairedsafety testingsickle erythroidsicklingtargeted treatmenttherapeutic targettool
项目摘要
PROJECT SUMMARY
Despite advances in the medical care of sickle cell disease (SCD), most patients continue to experience severe
pain, poor quality of life, progressive organ deterioration and premature death. Allogeneic hematopoietic stem
cell transplantation (HSCT) can cure SCD but is associated with numerous toxicities and only 20% of patients
have Human Leukocyte Antigen (HLA)-matched donors. Therefore, improved and more widely accessible
curative therapies are needed. Genetic modification of autologous HSCs is a promising experimental approach
for treating SCD that circumvents some of the problems associated with allogeneic HSCT, although the optimal
technical strategies are not yet established. This proposal explores the use of adenosine base editors (ABEs)
and prime editors (PEs) for genetic correction of SCD. In contrast to conventional genome editing, these novel
approaches create precise nucleotide alterations independent of double-stranded DNA breaks (DSBs), which
can cause structural DNA abnormalities, cell death or malignant transformation. Adenosine base editors convert
targeted A·T base pairs to G·C pairs. Prime editors copy edited sequence information from a guide RNA template
into a targeted DNA locus. We will test these potentially transformative tools in 3 different strategies for SCD
therapy. Aim 1 employs ABEs to create HSC alterations that recapitulate hereditary persistence of fetal
hemoglobin (HPFH), a benign genetic condition that alleviates the pathophysiology of co-inherited SCD by
inducing the expression of red blood cell (RBC) fetal hemoglobin (HbF), a potent anti-sickling agent. We have
used protein evolution strategies to create new high-efficiency ABEs that generate HPFH mutations at
frequencies of up to 60% in CD34+ hematopoietic stem and progenitor cells (HSPCs), with HbF being induced
to levels that inhibit hypoxic sickling of erythroid progeny. Aim 2 uses ABEs to convert the mutant SCD codon
from valine to alanine, thereby generating “Hemoglobin Makassar (HbG)”, a naturally occurring benign non-
sickling variant. We have developed an altered PAM-specific ABE that converts HbS alleles to HbG in SCD
donor HSPCs at frequencies of up to 80%, with inhibition of RBC sickling. Aim 3 employs prime editing to revert
the mutant SCD codon to normal (Val→Glu), which we have shown to occur efficiently in the HEK293T cell line
and now aim to optimize in HSPCs from affected individuals. Overall, our preliminary studies have shown proof
of principle for three novel, independent editing approaches to treating SCD without the need to enrich for edited
cells or to create DSBs. Through the proposed research, we seek to optimize the efficiency of these approaches
in primary HSPCs and to further determine their safety and efficacy by using mouse models, in vitro culture
methods and biochemical assays. Developing three approaches simultaneously will enable us to compare their
outcomes directly and to determine the best therapeutic strategy to pursue in future clinical studies. More
generally, our planned studies have the potential to generate new paradigms for using base editors and PEs to
treat numerous genetic blood disorders via precise genetic manipulation of HSCs.
项目总结
项目成果
期刊论文数量(0)
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{{ truncateString('DAVID R LIU', 18)}}的其他基金
Project 3: Therapeutic Gene Editing for Huntington's Disease
项目3:亨廷顿病的治疗性基因编辑
- 批准号:
10668769 - 财政年份:2023
- 资助金额:
$ 77.71万 - 项目类别:
Project 2: Therapeutic Gene Editing for Friedreich's Ataxia
项目 2:弗里德赖希共济失调的治疗性基因编辑
- 批准号:
10668768 - 财政年份:2023
- 资助金额:
$ 77.71万 - 项目类别:
Base editing and prime editing for sickle cell disease
镰状细胞病的碱基编辑和引物编辑
- 批准号:
10157511 - 财政年份:2021
- 资助金额:
$ 77.71万 - 项目类别:
Continuous Evolution of Proteins with Novel Therapeutic Potential
具有新治疗潜力的蛋白质的不断进化
- 批准号:
10181559 - 财政年份:2021
- 资助金额:
$ 77.71万 - 项目类别:
Base editing and prime editing for sickle cell disease
镰状细胞病的碱基编辑和引物编辑
- 批准号:
10323054 - 财政年份:2021
- 资助金额:
$ 77.71万 - 项目类别:
Continuous Evolution of Proteins with Novel Therapeutic Potential
具有新治疗潜力的蛋白质的不断进化
- 批准号:
10588186 - 财政年份:2021
- 资助金额:
$ 77.71万 - 项目类别:
Continuous Evolution of Proteins with Novel Therapeutic Potential
具有新治疗潜力的蛋白质的不断进化
- 批准号:
10393666 - 财政年份:2021
- 资助金额:
$ 77.71万 - 项目类别:
PedGeneRx - Admin Supplement to Base Editing and Prime Editing for Sickle Cell Disease R01
PedGeneRx - 镰状细胞病 R01 碱基编辑和 Prime 编辑的管理补充
- 批准号:
10594247 - 财政年份:2021
- 资助金额:
$ 77.71万 - 项目类别:
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