Recombinant AAV gene therapies deliver durable clinical benefit but face immune-mediated challenges that vary by vector, dose, route, and patient. Gulve and colleagues synthesize the clinical manifestations, temporal patterns, and mechanisms of rAAV immunogenicity, highlighting risk assessment and emerging mitigation strategies to support safer, more effective gene therapy development.

Infusion-related reactions (IRRs) can occur with AAV gene therapy. IRRs were observed in two participants who received AAV8 gene therapy and in one who received AAV9 gene therapy. AAV gene therapy IRRs may be rate-related, possibly driven by complement activation. In our studies, a slow, staged infusion mitigated additional IRRs.

Transduction efficiency is a critical determinant of CAR T-cell manufacturing quality. Analysis of 204 clinical CAR T-cell products revealed that transduction efficiency is shaped primarily by manufacturing workflows and protocol-dependent starting material composition. Higher transduction efficiency was associated with early memory-like cellular states, providing insights into optimizing CAR T-cell.

Automated ÄKTA-connected zonal-rotor CsCl ultracentrifugation enables high-resolution AAV polishing at 1.65-L scale. The workflow enriched full capsids and resolved empty and intermediate populations in AAV8-X and AAV9-Y. The AAV8-X intermediate fraction retained about a half of qPCR signal of equimolar full capsid but produced minimal mRNA expression potency.

Inoue and colleagues developed Digenome-Detect, a data analysis tool that enables highly accurate and sensitive assessment of off-target genome editing using Digenome-seq, the simplest cell-free assay. This advance improves genome-wide identification of off-target mutation events and thereby helps ensure the safety of genome-editing therapeutics.

Maeda and colleagues established of a 293-based cell line stably maintaining the HSV genome. This cell line enables efficiently producing HSV amplicon vectors without purification or transfection of the HSV genome. This system offers a more practical platform for HSV-based gene delivery, overcoming several limitations associated with conventional methods.