Jul 2026· International Journal of Molecular Sciences· Vol 27· 0 citations· 119 references
Medicine
TL;DR
Evidence that cell-type-specific promoters lose fidelity when paired with neurogenic transgene payloads and that published in vivo reprogramming efficiencies may be substantially confounded by promoter leakage in the absence of formal lineage tracing are synthesized.
Abstract
The adeno-associated virus (AAV) has become the vector of choice for gene therapy and experimental gene delivery, owing to its non-pathogenic nature and ability to achieve persistent gene expression across diverse tissues. AAV has emerged as a key platform in cellular reprogramming and regenerative medicine, with applications spanning transcription factor delivery for in vivo lineage conversion and tissue repair across the CNS, heart, and musculoskeletal systems. However, significant limitations remain, particularly in the context of induced pluripotent stem cell (iPSC) engineering. We assess barriers to efficient iPSC transduction including receptor-dependent entry deficits and activation of p53-dependent DNA damage responses. Although AAV is widely described as non-integrating, evidence indicates that integration events occur in rapidly proliferating and actively reprogramming cells. Critically, we synthesize evidence that cell-type-specific promoters lose fidelity when paired with neurogenic transgene payloads, a cross-tissue problem not addressed in existing AAV reviews, and that published in vivo reprogramming efficiencies may be substantially confounded by promoter leakage in the absence of formal lineage tracing. These aspects, underrepresented in recent platform-level reviews, are specifically emphasized here as a resource for researchers designing rigorous AAV-based reprogramming and gene therapy strategies.
Innovations in pericardial delivery systems and optimized manufacturing protocols now pave the way for future clinical translation of direct cardiac reprogramming strategies, which could redefine heart failure therapy.
Boram Son, Gwang Yeol Park, Ildoo Jeong et al.· Materials Today Bio· 0 citations
This review summarizes the trajectory of iPSC reprogramming technologies and identifies the core “translational triltrilas”, namely, the inherent tradeoffs between security, homogeneity, and scalability, and proposes a comprehensive strategy to overcome these bottlenecks.
Mengmeng Chen, Ning Zuo, Qi Wang et al.· Frontiers in Cell and Develo...· 0 citations
This review discusses how organoids and OoC-based platforms are being leveraged to study and enhance cell fate reprogramming, repair, and regeneration across multiple tissues and outlines current limitations, including scalability, standardization, and biomaterial constraints.
Hrithiha Sriramulu, Hyunsung Woo, Anavi Kaul et al.· Current Opinion in Genetics...· 1 citation
Several viral vectors have been developed for gene therapy due to their high transduction efficiency, but some integrate into the host genome, raising safety concerns. Recent studies have identified recombinant adeno-associated virus serotype 6 (rAAV6) as a promising vector for hematopoietic stem and progenitor cell-targeted gene therapy because of its non-pathogenic nature, low integration frequency, and capacity for sustained episomal transgene expression. Nevertheless, its chromosomal integration profile remains incompletely defined, warranting a comprehensive evaluation to assess long-term safety. In this study, human CD34＋ cells were transduced with rAAV6 under varying vector doses and transgene contexts, and integration-site mapping was performed using the integration-site enriched library sequencing approach. Consistent with the largely episomal nature of AAV, high vector sequence alignment rates were observed across all groups. rAAV6 integrations occurred randomly throughout the genome, showing a broad pan-chromosomal distribution without evidence of sequence-specific targeting or clustering. Although integrations were more frequent in CpG islands, commonly located within open chromatin, this pattern likely reflects chromatin accessibility rather than targeting bias. Functional enrichment analysis indicated associations with general cellular and structural processes, without enrichment in oncogenic pathways. Distance-based analysis confirmed that integration sites were mapped at a distance from oncogenes and tumor suppressor genes, even under high-dose conditions. The data support the genomic safety of rAAV6 and its applicability to hematological gene therapy.
H. Lee, Nayoung Park, In-Byung Park et al.· International Journal of Ste...· 0 citations
This review provides a comprehensive overview of the current status of viral and non-viral vector systems for in vivo and ex vivo applications, and key comparisons are made across safety, efficacy, scalability, and immune responses.