TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage DOI Open Access
Eduardo Seclén, Jessica Jang,

Aminah O. Lawal

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 5, 2024

Abstract Gene therapy in hematopoietic stem and progenitor cells (HSPCs) shows great potential for the treatment of inborn metabolic diseases. Typical HSPC gene approaches rely on constitutive promoters to express a therapeutic transgene, which is associated with multiple disadvantages. Here, we propose novel promoter-less intronic editing approach that triggers transgene expression only after cellular differentiation into myeloid lineage. We integrated splicing-competent eGFP cassette first intron CD11b observed lineage but minimal no HSPCs or differentiated non-myeloid lineages. In vivo , edited successfully engrafted immunodeficient mice displayed compartment tissues. Using same approach, expressed alpha-L-iduronidase (IDUA), defective enzyme Mucopolysaccharidosis type I, 10-fold supraendogenous IDUA exclusively differentiation. Edited efficiently populated bone marrow, blood, spleen mice, retained capacity secrete ex . Importantly, transgenes were also found brain. This may unlock new strategies neurological diseases require delivery therapeutics

Language: Английский

CRISPR-Based Gene Therapies: From Preclinical to Clinical Treatments DOI Creative Commons
Marine Laurent, Marine Geoffroy, Giulia Pavani

et al.

Cells, Journal Year: 2024, Volume and Issue: 13(10), P. 800 - 800

Published: May 8, 2024

In recent years, clustered regularly interspaced short palindromic repeats (CRISPRs) and CRISPR-associated (Cas) protein have emerged as a revolutionary gene editing tool to treat inherited disorders affecting different organ systems, such blood muscles. Both hematological neuromuscular genetic benefit from genome approaches but face challenges in their clinical translation. The ability of CRISPR/Cas9 technologies modify hematopoietic stem cells ex vivo has greatly accelerated the development therapies for disorders. last decade, many trials were initiated are now delivering encouraging results. FDA approval Casgevy, first CRISPR/Cas9-based drug severe sickle cell disease transfusion-dependent β-thalassemia, represents significant milestone field highlights great potential this technology. Similar preclinical efforts currently expanding CRISPR other hematologic primary immunodeficiencies. field, versatility been instrumental generation new cellular animal models Duchenne muscular dystrophy (DMD), offering innovative platforms speed up therapeutic solutions. Several corrective interventions proposed genetically restore dystrophin production using toolbox demonstrated promising results DMD models. Although these advances represent step forward translation DMD, there still hurdles overcome, delivery methods associated with high viral vector doses, together safety immunological concerns. Collectively, obtained fields emphasize transformative impact patients affected by debilitating conditions. As each suffers specific challenges, may progress differentially depending on disorder. Ongoing investigations will address risks limitations therapies, including long-term efficacy, genotoxicity, adverse immune reactions. This review provides insights into diverse applications CRISPR-based both settings monogenic compare while highlighting current trends, difficulties, overcome.

Language: Английский

Citations

15
Gene therapy for inborn errors of immunity: current clinical progress DOI
Sathi Wijeyesinghe, Javier Chinen

Annals of Allergy Asthma & Immunology, Journal Year: 2025, Volume and Issue: unknown

Published: March 1, 2025

Language: Английский

Citations

1
A p38 MAPK-ROS axis fuels proliferation stress and DNA damage during CRISPR-Cas9 gene editing in hematopoietic stem and progenitor cells DOI Creative Commons
Lucrezia della Volpe,

Federico Midena,

Roberta Vacca

et al.

Cell Reports Medicine, Journal Year: 2024, Volume and Issue: 5(11), P. 101823 - 101823

Published: Nov. 1, 2024

Ex vivo activation is a prerequisite to reaching adequate levels of gene editing by homology-directed repair (HDR) for hematopoietic stem and progenitor cell (HSPC)-based clinical applications. Here, we show that shortening culture time mitigates the p53-mediated DNA damage response CRISPR-Cas9-induced double-strand breaks, enhancing reconstitution capacity edited HSPCs. However, this results in lower HDR efficiency, rendering ex necessary yet detrimental. Mechanistically, triggers multi-step process initiated p38 mitogen-activated protein kinase (MAPK) phosphorylation, which generates mitogenic reactive oxygen species (ROS), promoting fast cell-cycle progression subsequent proliferation-induced damage. Thus, inhibition before delays G1/S transition expands transcriptionally defined HSCs, ultimately endowing cells with superior multi-lineage differentiation, persistence throughout serial transplantation, enhanced polyclonal repertoire, better-preserved genome integrity. Our data identify proliferative stress as driver HSPC dysfunction fundamental implications designing more effective safer correction strategies

Language: Английский

Citations

5
Rediscovering the human thymus through cutting-edge technologies DOI Creative Commons
Francesca Pala, Luigi D. Notarangelo, Marita Bosticardo

et al.

The Journal of Experimental Medicine, Journal Year: 2024, Volume and Issue: 221(10)

Published: Aug. 21, 2024

Recent technological advances have transformed our understanding of the human thymus. Innovations such as high-resolution imaging, single-cell omics, and organoid cultures, including thymic epithelial cell (TEC) differentiation culture, improvements in biomaterials, further elucidated thymus architecture, cellular dynamics, molecular mechanisms underlying T development, unraveled previously unrecognized levels stromal heterogeneity. These advancements offer unprecedented insights into biology hold promise for development novel therapeutic strategies immune-related disorders.

Language: Английский

Citations

4
Gene Therapy Strategies for RAG1 Deficiency: Challenges and Breakthroughs DOI Creative Commons
Giorgio Gilioli, Arjan C. Lankester, Sander de Kivit

et al.

Immunology Letters, Journal Year: 2024, Volume and Issue: unknown, P. 106931 - 106931

Published: Sept. 1, 2024

Language: Английский

Citations

4
Gene Therapy for Inborn Errors of Immunity DOI
Jean‐Sébastien Diana, Marina Cavazzana

Elsevier eBooks, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

Language: Английский

Citations

0
Potential Genetic Approach to Specific Primary Immunodeficiencies: Which Perspectives? DOI Creative Commons
Giuseppe Murdaca, Francesca Paladin, Sebastiano Gangemi

et al.

Frontiers in Bioscience-Landmark, Journal Year: 2025, Volume and Issue: 30(3)

Published: March 17, 2025

IMR Press is a leading publisher of open access peer-reviewed biomedical and life sciences journals. We aim to facilitate the dissemination high-quality research in area science. With long tradition wide readership, dedicated making positive contributions academics, corporate libraries as well readers authors. All editors will provide best service for researchers, allowing them have easy smooth publication experience helping maximize impact visibility their research.

Language: Английский

Citations

0
Organoid Models of Lymphoid Tissues DOI Creative Commons
Ania Bogoslowski, Jun Ren, Clément Quintard

et al.

Organoids, Journal Year: 2025, Volume and Issue: 4(2), P. 7 - 7

Published: April 7, 2025

Lymphoid organs are critical for organizing the development of immune system, generating tolerance, and orchestrating adaptive response to foreign antigens. Defects in their structure function can lead immunodeficiency, hypersensitivity, cancer, or autoimmune diseases. To better understand these diseases assess potential therapies, complex models that recapitulate anatomy physiology tissues required. Organoid possess a number advantages, including 3D microarchitecture, scalability, personalization, which make them ideal modelling lymphoid related pathologies. Organoids have been developed both primary secondary tissues; however, several limitations, immature phenotypes incomplete stromal cell populations. Furthermore, organoids often heterogeneous function. Several organs, such as spleen, do not yet robust organoid models, offering opportunities breakthroughs field. Overall, will pave way rapid testing novel organ modelling, personalized medicine. This review summarizes current advances organ—bone marrow thymus—as well lymph node spleen.

Language: Английский

Citations

0
TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage DOI
Eduardo Seclén, Jessica Jang,

Aminah O. Lawal

et al.

Molecular Therapy, Journal Year: 2024, Volume and Issue: 32(6), P. 1643 - 1657

Published: April 6, 2024

Language: Английский

Citations

2
TALEN-mediated intron editing of HSPCs enables transgene expression restricted to the myeloid lineage DOI Open Access
Eduardo Seclén, Jessica Jang,

Aminah O. Lawal

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: March 5, 2024

Abstract Gene therapy in hematopoietic stem and progenitor cells (HSPCs) shows great potential for the treatment of inborn metabolic diseases. Typical HSPC gene approaches rely on constitutive promoters to express a therapeutic transgene, which is associated with multiple disadvantages. Here, we propose novel promoter-less intronic editing approach that triggers transgene expression only after cellular differentiation into myeloid lineage. We integrated splicing-competent eGFP cassette first intron CD11b observed lineage but minimal no HSPCs or differentiated non-myeloid lineages. In vivo , edited successfully engrafted immunodeficient mice displayed compartment tissues. Using same approach, expressed alpha-L-iduronidase (IDUA), defective enzyme Mucopolysaccharidosis type I, 10-fold supraendogenous IDUA exclusively differentiation. Edited efficiently populated bone marrow, blood, spleen mice, retained capacity secrete ex . Importantly, transgenes were also found brain. This may unlock new strategies neurological diseases require delivery therapeutics

Language: Английский

Citations

0