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.

He and colleagues developed a transient, non-gene-editing platform for off-the-shelf allogeneic TCR-T therapy. By conferring tacrolimus resistance characteristics to IL-2/4/7 expanded TCR-T cells that display reduced alloreactivity, He et al. provided a proof-of-concept for an allogeneic TCR-T therapy approach that addresses both manufacturability, and unwanted graft and host alloreactivity concerns.

In 2012, I had the distinct honor of being part of one of the trailblazing teams leading first-in-human/first-in-child chimeric antigen receptor (CAR) T cell trials in B cell acute lymphoblastic leukemia (B-ALL). Little did I know then that we were only at the tip of the iceberg for use of immunotherapy in pediatric oncology. While the multiple CAR T cell approvals for B cell malignancies, including in pediatric B-ALL, reflect the curative potential in relapsed/refractory disease, what has additionally transpired is a cascade of novel immune-effector cell therapy-based approaches, each iteratively building upon the prior to further extend the therapeutic index of immunotherapy.

Zhang and colleagues engineered the poly(A) tail as a programmable regulatory element, showing that embedding cell-type-specific microRNA target sites directly within it confers robust, position-dependent silencing in off-target tissues while preserving activity in target cells. This strategy offers a new modular tool to enhance mRNA therapeutic safety and precision.

Kis and colleagues report an integrated sequential-batch IVT–oligo-dT process that links RNA synthesis and affinity capture through a shared buffer, enabling direct crude-IVT loading and flowthrough recycling. The workflow improves cap-analog utilization and raw-material efficiency while preserving functional mRNA expression across five cycles.

COL17A1 nonsense mutations generate premature termination codons, reducing collagen XVII through truncated protein production and/or nonsense-mediated decay (NMD), and causing junctional epidermolysis bullosa. Translational readthrough drug offers a potential approach to nonsense mutations suppression. Different COL17A1 nonsense mutations showed distinct responses to readthrough drugs and NMD inhibitors, supporting personalized therapeutic strategies.

CRISPR hybrid guides containing RNA and DNA nucleotides (chRDNA) enhance Cas nuclease specificity by reducing off-target editing. Intravenous administration of LNP-encapsulated Cas12a mRNA with a chRDNA guide in mice led to dose-responsive hepatic genome editing without detectable off-target editing and near elimination of the targeted plasma protein for one year.

Immunization with a nucleoside-modified mRNA-LNP vaccine encoding the conserved malaria antigen E140 induced durable humoral immunity in mice and generated invasion-blocking antibodies against Plasmodium vivax. The results highlight E140 as a promising candidate for next-generation malaria vaccines.

Lipid nanoparticles incorporating DOTAP and DOPE enable serial mRNA transfection of primary human T cells with significantly improved viable cell yields compared to standard electroporation. This platform supports iterative functional modifications across multiple rounds of delivery while preserving cell viability, facilitating the manufacturing of complex, multi-payload cell therapies.

Ionizable lipid components of LNP vaccine formulations play an integral role in providing the necessary immune stimulatory signals required for effective anti-tumor T cell responses. This study finds that inclusion of a novel ionizable lipid confers effective tumor control in part through the generation of type I interferons.

Gene therapy is transforming hereditary hematological disorders. This review summarizes approved gene addition, editing, and silencing strategies for sickle cell disease, thalassemia, and hemophilia, highlights curative potential, and discusses remaining challenges such as immune responses, cost, and accessibility

Circular RNAs (circRNAs) have emerged as promising next-generation therapeutic platforms due to their exceptional stability, prolonged protein expression, and low immunogenicity. This review highlights advances in enzymatic, chemical, and bio-orthogonal circRNA synthesis, compares their advantages and limitations, and discusses emerging applications in vaccines, cancer therapy, and precision medicine.

(Molecular Therapy 31, 1596–1614; June 2023)

Yuan and colleagues demonstrate that inhaled biodegradable nanosilica activates an alveolar macrophage–NK axis, triggering an IL-12/15/18 triad that programs memory-like NK cells. This non-fibrotic, cell-free strategy suppresses melanoma growth, prevents postsurgical recurrence, and synergizes with anti-PD-1, establishing a robust framework for in vivo NK cell immunotherapy.

Zhang and colleagues reveal that GPX4 serves as a critical regulator of NRF2-mediated otoprotection against aminoglycoside-induced hair cell injury. Their findings identify a GPX4-dependent antioxidant mechanism that enables therapeutic activation of NRF2 and provides new insights into strategies for preventing drug-induced hearing loss.

Lu and colleagues identify BRD4 as a central epigenetic driver of vesicant-induced corneal injury. Using reproducible mouse and rabbit models, they show that short-term topical BRD4 inhibition suppresses acute inflammation and provides durable protection of corneal clarity, stromal organization, endothelial integrity, and neovascularization, supporting translational therapy for chemical eye injuries.

The authors develop AminiCRoff, a compact dAsCas12f1-based epigenetic silencer compatible with single-AAV delivery. AminiCRoff mediates durable, heritable gene silencing comparable to the larger CRISPRoff and represses endogenous MYC to inhibit tumor cell proliferation, providing a versatile platform for in vivo epigenome editing.

Blancafort and colleagues describe a non-viral polymeric system for the delivery of dCas9-KRAB silencers as ribonucleoprotein (RNP) payloads for EWSR1-FLI1 repression. They demonstrate highly efficient RNP delivery and robust silencing of EWSR1-FLI1 in both cell line and patient-derived xenografts of Ewing sarcoma, accompanied by potent anti-tumor effects.