Development of an Usher Gene Therapy

Usher 基因疗法的开发

基本信息

批准号：
10343826
负责人：
Uwe D Staerz
金额：
$ 11万
依托单位：
GREFFEX, INC.
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：
2021-03-01 至 2023-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10343826
关键词：
Address Adenoviruses Affect Animals Area Benign Blindness CRISPR/Cas technology Capsid Cell Differentiation process Cells Clinic Clinical Code Collaborations Complementary DNA DNA Defect Dependovirus Development Engineering Exons Eye Future Gene Transfer Genes Genetic Genome Green Fluorescent Proteins Helper Viruses Human Immunologist In Situ In Vitro Inflammatory Inflammatory Response Injections Investigation Knockout Mice Length Lentivirus Lentivirus Vector Molecular Mutation National Heart, Lung, and Blood Institute Nature Organ Pathogenicity Phase Photoreceptors Population Replication-Associated Process Retina Retinal Degeneration Retinitis Pigmentosa Serotyping Small Business Innovation Research Grant Symptoms System Technology Testing Therapeutic Time Toxic effect Transgenes USH2A gene United States National Institutes of Health Universities Usher Syndrome Type 2 Utah Virus Virus Replication Yang base deafness design dimer effectiveness testing exome sequencing exon skipping experience eye center gene repair gene therapy gene transfer vector gutless adenoviral vector hearing impairment immunogenicity in vivo innovation moran A nanoparticle novel photoreceptor degeneration promoter repaired symposium transgene expression vector vector control

Address Adenoviruses Affect Animals Area Benign Blindness CRISPR/Cas technology Capsid Cell Differentiation process Cells Clinic Clinical Code Collaborations Complementary DNA DNA Defect Dependovirus Development Engineering Exons Eye Future Gene Transfer Genes Genetic Genome Green Fluorescent Proteins Helper Viruses Human Immunologist In Situ In Vitro Inflammatory Inflammatory Response Injections Investigation Knockout Mice Length Lentivirus Lentivirus Vector Molecular Mutation National Heart, Lung, and Blood Institute Nature Organ Pathogenicity Phase Photoreceptors Population Replication-Associated Process Retina Retinal Degeneration Retinitis Pigmentosa Serotyping Small Business Innovation Research Grant Symptoms System Technology Testing Therapeutic Time Toxic effect Transgenes USH2A gene United States National Institutes of Health Universities Usher Syndrome Type 2 Utah Virus Virus Replication Yang base deafness design dimer effectiveness testing exome sequencing exon skipping experience eye center gene repair gene therapy gene transfer vector gutless adenoviral vector hearing impairment immunogenicity in vivo innovation moran A nanoparticle novel photoreceptor degeneration promoter repaired symposium transgene expression vector vector control

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项目摘要

SUMMARY Mutations in the USH2A gene are the cause of the Usher syndrome type II (USH2) that is characterized by retinitis pigmentosa (RP) and hearing impairments. USH2 is the leading genetic cause of combined blindness and deafness. It affects about 400,000 subjects worldwide. There is no cure or therapy for USH2A- associated photoreceptor degeneration. The most common USH2A mutations are located in in-frame exon 13. Skipping of exon 13 has been investigated as therapy, even though a clinical observation suggested that exon 13 skipping may be pathogenic. Gene repair using the CRISPR/Cas9 machinery represents another approach to correct exon 13 mutations. Although highly efficient ex vivo after enrichment for corrected cells, it is not known whether the in situ homology-directed repair of possibly 25% of cells will suffice. A gene transfer vector controls its efficacy by its transduction rate. It addresses all pathogenic mutations within the entirety of the USH2A gene (>470 mutations, NHLBI/NIH Exome Sequencing Project). The USH2A coding cDNA is 15.6kb in length. It cannot be accommodated by conventional adeno-associated virus (AAV) and lentivirus vectors, whose payloads are limited to 5kb and 10kb, respectively. We have developed a fully deleted adenoviral vector (GreGT) that avoids contaminations with helper viruses and replication competent adenoviruses (RCA). With a payload of up to 33kb, we hypothesize that it will efficiently deliver a full-length USH2A coding cDNA to its photoreceptor targets. We expect that these studies will establish the GreGT vector as platform for the treatment of other defects afflicting large genes. Our application builds upon a collaboration between UD Staerz (Greffex, Inc.), who as an experienced immunologist and molecular biologist has guided the development of different vector systems, and Jun Yang (John A Moran Eye Center, University of Utah), who has developed a unique expertise in the area of retinal degeneration.

概括 USH2A基因中的突变是USHER综合征II型（USH2）的原因，其特征是色素性视网膜炎（RP）和听力障碍。 USH2是结合失明的主要遗传原因和耳聋。它影响了全球约40万受试者。没有治疗或治疗USH2A-- 相关的光感受器变性。最常见的USH2A突变位于框架外显子 13.外显子13的跳过已被研究为治疗，尽管临床观察表明外显子跳过可能是致病性的。使用CRISPR/CAS9机械的基因维修代表另一种纠正外显子13突变的方法。虽然富集在校正细胞后的效率高度效率尚不清楚可能25％的细胞的原位同源性修复是否足够。基因转移向量通过其转导率控制其疗效。它解决了整个中的所有致病突变 USH2A基因（> 470个突变，NHLBI/NIH外显子组测序项目）。 USH2A编码cDNA是长度为15.6kb。传统的腺相关病毒（AAV）和慢病毒无法容纳它向量分别限制为5KB和10KB。我们已经开发了一个完全删除的腺病毒载体（GREGT）避免了辅助病毒和复制能力的污染腺病毒（RCA）。有效载荷最多为33kb，我们假设它将有效地提供全长 USH2A将cDNA编码为其光感受器靶标。我们希望这些研究将建立Gregt载体作为治疗其他缺陷的平台。我们的应用程序建立在 UD Staerz（Greffex，Inc。）之间的合作，他是经验丰富的免疫学家和分子生物学家指导了不同的向量系统的发展，Jun Yang（John Moran眼中心，犹他大学），他在视网膜变性领域建立了独特的专业知识。