Molecular Therapy-Methods & Clinical Development最新文献

Scientific breakthroughs in the field of cell and gene therapy lay the groundwork for therapeutic innovation, but their translation and integration into routine care is delayed by systemic inefficiencies and limited cross-sector alignment. The Advanced Therapies model of Lund is a proactive and integrated approach designed to accelerate the development of advanced therapies from discovery to reimbursement, facilitating close collaboration among three main entities: university, hospital, and the iActors (a coalition of the innovation system, incubators, investors, and industry). Unlocking the full potential of therapeutic innovation requires inclusive and proactive engagement from all entities to accelerate development timelines, optimize resource use across the ecosystem, and ultimately deliver curative, cost-effective advanced therapies to patients. This will ensure broad patient access and establishment of optimal conditions for developing the next generation of cell and gene therapies, while also facilitating innovation and economic growth. To operationalize this model, we developed the Cell and Gene Therapy Navigator, a visualization instrument designed to track and optimize the development of advanced therapies within the model framework. This tool leverages three-dimensional visualization to provide a dynamic and comprehensive overview of progress of the advanced therapy development across the three crucial entities: university, hospital, and iActors. We are confident that working according to the model will minimize translational gaps and result in seamless good manufacturing practices (GMP) transfer, fewer re-works, faster time to first-in-human, and the creation of more viable biotech companies.

[This corrects the article DOI: 10.1016/j.omtm.2019.11.016.].

Eliminating hepatitis B virus (HBV) covalently closed circular DNA (cccDNA) remains a major challenge, requiring innovative treatment strategies and drug candidates. Clinical studies reveal that wild-type HBV in the blood is often replaced by gradually rising mutant populations. This replacement reflects loss of the early cccDNA pool, then replenished predominantly through de novo infection. We proposed that blocking de novo infection is essential for cccDNA elimination and establishing a finite HBV treatment regimen. To achieve sustained inhibition of de novo cccDNA replenishment, we developed HBVZ10, a gene therapy candidate that utilizes an optimized adeno-associated virus (AAV) vector 8 to deliver human anti-HBs antibody genes into muscle cells for expanding endogenous anti-HBs antibody production capacity. HBVZ10 expression and therapeutic function were evaluated in uPA/SCID chimeric mice. HBVZ10 therapy achieved sustained antibody expression of ≥100,000 mIU/mL for at least 200 days following a single dose administration. Combining HBVZ10 with intracellular replication inhibitor entecavir resulted in >100-fold reductions in cccDNA within a few months, accompanied by progressive reductions in serum HBeAg and HBsAg to undetectable levels. These findings establish preclinical evidence of HBVZ10 as a novel gene therapy candidate and support a paradigm-shifting cccDNA elimination strategy.

Adoptive T cell therapy, particularly T cell receptor-engineered T (TCR-T) cell therapy, holds promise for cancer treatment in solid tumors and hematological malignancies. Conventional lentiviral TCR-T cell therapies face insertional mutagenesis risks, while CRISPR-mediated T cell receptor α constant (TRAC) locus targeting suffers from suboptimal knockin efficiency and chromosome 14 loss. To address these challenges, we introduced a non-viral strategy combining CRISPR-Cas9 electroporation with methotrexate (MTX) metabolic selection. Primary human T cells were engineered to integrate a CMV-pp65-specific TCR and an MTX-resistant dihydrofolate reductase (DHFR)-FS cassette into the TRAC locus via homology-directed repair (HDR). Systematic optimization of electroporation timing, buffer systems, and HDR enhancers achieved initial TCR integration efficiency of ∼20%. Subsequent 6-day MTX treatment enriched engineered cells to ∼70% purity while selectively depleting unedited and chromosomally aberrant clones. Fluorescence in situ hybridization revealed that MTX enrichment reduced CRISPR-associated chromosome 14 loss comparable to unedited T cells. Functionally, TRAC-TCR-T cells exhibited enhanced interferon (IFN)-γ/tumor necrosis factor alpha (TNF-α) secretion and reduced exhaustion markers versus lentiviral counterparts, while maintaining equivalent tumor clearance efficacy in vitro and in xenograft models. In conclusion, this integrated platform mitigates viral vector risks, alleviates concerns of CRISPR-associated genomic instability, and provides a good manufacturing practice (GMP)-compatible approach that may facilitate the development of safer adoptive TCR-T immunotherapies.