Lentiviral Gene Therapy and Genome Editing for Wiskott-Aldrich Syndrome

Wiskott-Aldrich 综合征的慢病毒基因治疗和基因组编辑

• 批准号: 9762186
• 负责人: David J Rawlings
• 金额: $ 62.16万
• 依托单位: ST. JUDE CHILDREN'S RESEARCH HOSPITAL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9762186
• 关键词: AMD3100; Adverse event; Animal Model; Animals; Atypical lymphocyte; Autoimmune Diseases; Autoimmunity; Autologous; Automobile Driving; B-Lymphocytes; Bacterial Infections; Biological Assay; Blood Platelets; Busulfan; CD34 gene; CRISPR/Cas technology; CSF3 gene; Cell Count; Cell Lineage; Cell physiology; Cells; Child; Cleaved cell; Clinical; Clinical Trials; Clonal Expansion; Clustered Regularly Interspaced Short Palindromic Repeats; Complementary DNA; Coupled; DNA; Data Set; Development; Disease; Eczema; Electroporation; Engineered Gene; Engraftment; Enhancers; Enrollment; Exhibits; Funding; Future; Gene Expression; Gene Targeting; Gene Transfer; Genes; Genome; Goals; Hematopoietic; Hematopoietic stem cells; Hemorrhage; Homing; Human; Immune; Immunologic Deficiency Syndromes; Immunologics; In Vitro; Infection; Insulator Elements; Lentivirus Vector; Link; Liquid substance; Lymphocyte Function; Malignant Neoplasms; Mediating; Messenger RNA; Methods; Monitor; Morbidity - disease rate; Multi-Institutional Clinical Trial; Mus; Mutagenesis; Myelogenous; Natural Killer Cells; Opportunistic Infections; Patients; Pattern; Pediatric Hospitals; Peripheral; Peripheral Blood Stem Cell; Phagocytes; Platelet Count measurement; Pre-Clinical Model; Production; Proteins; Protocols documentation; Publishing; Reagent; Recovery; Regulation; Regulatory T-Lymphocyte; Research Institute; Resolution; Risk; Safety; Saint Jude Children' s Research Hospital; Serious Adverse Event; Sickle Cell Anemia; Site; Stem cells; System; T-Lymphocyte; Testing; Thrombocytopenia; Toxic effect; Transplantation; Treatment Protocols; United States National Institutes of Health; Viral; Virus Diseases; Wiskott-Aldrich Syndrome; Work; base; cellular transduction; clinical application; clinically translatable; cohort; conditioning; curative treatments; cytotoxicity; design; efficacy testing; endonuclease; engineered nucleases; fludarabine; gene product; gene replacement; gene therapy; gene therapy clinical trial; genome editing; immune function; in vivo; innate immune function; integration site; lentivirally transduced; leukemia; next generation; nonhuman primate; novel; nuclease; peripheral blood; pre-clinical; preclinical study; programs; promoter; protein expression; repaired; safety and feasibility; stem cell differentiation; therapeutic gene; tool; transcription activator-like effector nucleases; vector

PROJECT SUMMARY – PROJECT 2 Dr. David Rawlings, PI, will direct overall activities in this project, including all work performed at Seattle Children’s Research Institute (SCRI) and coordination of work performed by our collaborating program sites. We will implement and participate in a novel lentiviral (LV)-based clinical gene therapy trial for patients with Wiskott- Aldrich Syndrome (WAS). This trial will test the CL20-i650-MND-huWAS LV vector. Clinical LV will be generated by St. Jude Children’s Research Hospital (St. Jude) using a stable producer clone. GMP LV stocks will be used to transduce G-CSF/plerixafor mobilized peripheral blood CD34+ cells from patients with WAS using a two-hit protocol. Transduced cells will be re-infused into the patient after subablative conditioning using fludarabine and targeted busulfan. We will enroll up to 15 total patients at our three study sites: Seattle Children’s Hospital, the NIH Clinical Center, and St. Jude. Overall, this trial will provide important new information regarding the use of LV to treat WAS, as well as other disorders requiring high-level therapeutic gene expression in multiple lineages. In parallel with this trial, we will perform WAS gene editing studies at Seattle. We will leverage our broad expertise in nuclease engineering and gene editing in primary cells to develop next-generation pre-clinical tools for WAS gene targeting. Co-delivery of donor template and mRNA encoding novel homing endonuclease, TALEN or CRISPR reagents will be used to edit the endogenous WAS locus or candidate safe-harbor sites. Following optimization in control CD34+ HSC in vitro, we will assess function in vivo following engraftment in NSG recipient mice. Finally, we will perform pre-clinical studies using HSC from WAS subjects.

项目总结-项目2