Journal article
Highly efficient in vivo hematopoietic stem cell transduction using an optimized self-complementary adeno-associated virus
Molecular therapy. Methods & clinical development, Vol.33(1), p.101438
03/13/2025
DOI: 10.1016/j.omtm.2025.101438
PMCID: PMC11930595
PMID: 40129926
Abstract
In vivo
gene therapy targeting hematopoietic stem cells (HSCs) holds significant therapeutic potential for treating hematological diseases. This study uses adeno-associated virus serotype 6 (AAV6) vectors and Cre recombination to systematically optimize the parameters for effective
in vivo
HSC transduction. We evaluated various genetic architectures and delivery methods of AAV6, establishing an optimized protocol that achieved functional recombination in more than two-thirds of immunophenotypic HSCs. Our findings highlight that second-strand synthesis is a critical limiting factor for transgene expression in HSCs, leading to significant under-detection of HSC transduction with single-stranded AAV6 vectors. We also demonstrate that HSCs in the bone marrow (BM) are readily accessible to transduction, with neither localized injection nor mobilization of HSCs into the bloodstream, enhancing transduction efficacy. Additionally, we observed a surprising preference for HSC transduction over other BM cells, regardless of the AAV6 delivery route. Together, these findings not only underscore the potential of AAV vectors for
in vivo
HSC gene therapy but also lay a foundation that can inform the development of both
in vivo
AAV-based HSC gene therapies and potentially
in vivo
HSC gene therapies that employ alternative delivery modalities.
C. T. Charlesworth and colleagues demonstrate high rates of HSC transduction
in vivo
by AAV6, an optimized AAV6 vector for
in vivo
HSC transduction, and that second-strand synthesis limits transgene expression from AAV6 vectors in HSCs. The findings underscore the potential of AAVs for
in vivo
HSC gene therapy.
Details
- Title: Subtitle
- Highly efficient in vivo hematopoietic stem cell transduction using an optimized self-complementary adeno-associated virus
- Creators
- Carsten T. Charlesworth - Stanford University School of MedicineShota Homma - Stanford University School of MedicineAnais K. Amaya - Stanford University School of MedicineCarla Dib - Stanford University School of MedicineSriram Vaidyanathan - Nationwide Children's HospitalTze-Kai Tan - Stanford University School of MedicineMasashi Miyauchi - Stanford University School of MedicineYusuke Nakauchi - Stanford University School of MedicineFabian P. Suchy - Stanford University School of MedicineSicong Wang - Stanford University School of MedicineKyomi J. Igarashi - Stanford University School of MedicineM. Kyle Cromer - University of California, San FranciscoAmanda M. Dudek - Stanford University School of MedicineAlvaro Amorin - Stanford University School of MedicineAgnieszka Czechowicz - Stanford University School of MedicineAdam C. Wilkinson - University of CambridgeHiromitsu Nakauchi - Stanford University School of Medicine
- Resource Type
- Journal article
- Publication Details
- Molecular therapy. Methods & clinical development, Vol.33(1), p.101438
- DOI
- 10.1016/j.omtm.2025.101438
- PMID
- 40129926
- PMCID
- PMC11930595
- NLM abbreviation
- Mol Ther Methods Clin Dev
- ISSN
- 2329-0501
- eISSN
- 2329-0501
- Publisher
- American Society of Gene & Cell Therapy
- Language
- English
- Date published
- 03/13/2025
- Academic Unit
- Microbiology and Immunology
- Record Identifier
- 9984823123702771
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