In vivo engineering of CAR immune cells by nonviral nanoparticles.

Nanomaterials Drive In Vivo CAR Immune Cells Engineering.

Chimeric antigen receptor (CAR) immune cell therapy has emerged as a cornerstone of modern cell-based medicine, demonstrating potent clinical efficacy against a range of malignant tumors and autoimmune diseases. Nevertheless, conventional ex vivo CAR immune cell manufacturing is hindered by complexity, high costs, and significant inter-individual variability, which have limited its broad clinical application. These bottlenecks have prompted a paradigm shift toward in vivo CAR engineering, wherein nanocarriers directly deliver genetic material to circulating or tissue-resident immune cells, substantially simplifying the production process. This review first outlines the evolution of CAR immune cell therapy and key limitations of ex vivo approaches and then examines major delivery platforms for in vivo approaches. Comparative analyses are presented across delivery efficiency, cellular tropism, and expression kinetics, with particular emphasis on delineating the mechanistic distinctions and application boundaries between transient and durable expression strategies. We further dissect the critical challenges, including receptor-mediated non-specific immune activation, nanocarrier-associated immunogenicity constraining repeated administration, and delivery barriers imposed by the solid tumor microenvironment. Finally, the intrinsic trade-off between therapeutic durability and biosafety across distinct delivery platforms is discussed, along with the necessity that their clinical translation hinges on systematic optimization of delivery precision, immune compatibility, and expression controllability.

In vivo CAR-T therapy: The shift from ex vivo culturing to direct in situ immune reprogramming.

This review highlights key engineering strategies enabling in vivo CAR T-cell generation, summarizes emerging clinical research and development, and discusses future opportunities for expanding in vivo CAR T-cell therapies as scalable immunotherapy platforms.

Transient T cell therapies.

Engineering the blood: Lipid nanoparticle platforms for ex vivo immune and hematopoietic cell therapies.

This review aims to provide actionable guidance to bench-side cell engineering to broad clinical applications by comprehensively discussing recent advancements in LNP formulation and next-generation RNA payload engineering.

Targeted delivery of mRNA to immune cells for in vivo cell therapy

Abstract Messenger RNA (mRNA) therapeutics have revolutionized biomedicine by enabling direct in vivo programming of immune cells. This strategy bypasses the complex manufacturing and high costs associated with ex vivo cell therapies. However, efficient and specific systemic delivery of mRNA to target immune cell subsets remains a major translational hurdle. This review systematically examines engineering strategies that address this challenge. We first outline the key biological barriers to mRNA delivery, such as serum instability, nonspecific biodistribution, cellular uptake heterogeneity, and inefficient endosomal escape. Next, we comprehensively review advances in lipid nanoparticle (LNP) engineering, including discovering novel lipids, modulating compositions, conjugating targeting ligands, and incorporating stimuli-responsive elements, to enable enhanced tropism toward specific immune cells. Representative applications in oncology, protein replacement, autoimmune disease, and tissue regeneration are highlighted. Finally, we address translational challenges in safety, scalable manufacturing, and regulatory issues. The integration of rational material design, high-throughput screening, artificial intelligence, and interdisciplinary collaboration will be essential to advance next-generation targeted in vivo mRNA cell therapies toward clinical translation.

Engineering T cells in vivo to express a synthetic cytokine receptor enables preferential expansion and enrichment of anti-CD22 CAR T cells.

Ex vivo-manufactured chimeric antigen receptor (CAR) T cells targeting CD19 have transformed treatment strategies for patients with B cell malignancies and shown promising results in autoimmune disease. Yet despite their success, significant barriers remain that impede patient access to these therapies, including their complex manufacturing and the need for lymphodepleting chemotherapy. Moreover, many patients who receive ex vivo-manufactured CD19 CAR T cells do not achieve durable responses, suggesting that the expansion and persistence of CAR T cells may need improvement, and/or that strategies targeting alternative antigens may be needed for patients who are refractory to or relapse following treatment with CD19-directed therapies. Umoja is developing a surface-engineered lentiviral vector (LVV) platform capable of generating CAR T cells in vivo without lymphodepletion. UB-VV400 delivers a payload comprised of a fully human anti-CD22 CAR and the rapamycin-activated cytokine receptor (RACR), which enable preferential expansion and enrichment of CAR T cells in response to rapamycin. Here, we describe preclinical studies demonstrating that UB-VV400 generates anti-CD22 CAR T cells that kill CD22-expressing B cells and tumor cells in vitro and in vivo, and that engagement of RACR results in the selective expansion and enrichment of CAR T cells in vivo leading to complete tumor clearance and B cell depletion.