Homology-Directed-Repair-Based Genome Editing in HSPCs for the Treatment of Inborn Errors of Immunity and Blood Disorders

Allen, Daniel; Kalter, Nechama; Rosenberg, Michael; Hendel, Ayal

doi:10.3390/pharmaceutics15051329

Cited by 7 publications

(1 citation statement)

References 142 publications

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“…locations 7 . Using a targeted or safe harbor approach a second generation of treatments for metabolic diseases could be on its way given the positive results obtained in many preclinical studies 8 . However, both lentiviral and targeted integration approaches generally rely on the use of ubiquitous promoters to drive transgene expression.…”

Section: Introductionmentioning

confidence: 99%

TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage

Seclen,

Jang,

Lawal

et al. 2024

Preprint

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Gene therapy in hematopoietic stem and progenitor cells (HSPCs) shows great potential for the treatment of inborn metabolic diseases. Typical HSPC gene therapy approaches rely on constitutive promoters to express a therapeutic transgene, which is associated with multiple disadvantages. Here, we propose a novel promoter-less intronic gene editing approach that triggers transgene expression only after cellular differentiation into the myeloid lineage. We integrated a splicing-competent eGFP cassette into the first intron ofCD11band observed expression of eGFP in the myeloid lineage but minimal to no expression in HSPCs or differentiated non-myeloid lineages.In vivo, edited HSPCs successfully engrafted in immunodeficient mice and displayed transgene expression in the myeloid compartment of multiple tissues. Using the same approach, we expressed alpha-L-iduronidase (IDUA), the defective enzyme in Mucopolysaccharidosis type I, and observed a 10-fold supraendogenous IDUA expression exclusively after myeloid differentiation. Edited cells efficiently populated bone marrow, blood, and spleen of immunodeficient mice, and retained the capacity to secrete IDUAex vivo. Importantly, cells edited with the eGFP and IDUA transgenes were also found in the brain. This approach may unlock new therapeutic strategies for inborn metabolic and neurological diseases that require the delivery of therapeutics in brain.

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Section: Introductionmentioning

confidence: 99%