Every entry summarises what the study set out to test, what it found and why it changes how we design delivery systems. Browse research and reviews or search the collection.
Hannah J Vaughan, Camila G Zamboni, Kathryn M Luly, Ling Li, Kathleen L Gabrielson, Laboni F Hassan, Nicholas P Radant, Pranshu Bhardwaj, Florin M Selaru, Martin G Pomper, Jordan J Green
Hepatocellular carcinoma (HCC) has limited treatment options, with modest survival after systemic chemotherapy or transarterial chemoembolization (TACE). Gene therapies hold promise for treating HCC, but delivery remains a critical hurdle. While poly(beta-amino ester) (PBAE) nanoparticles have shown efficacy in transfecting HCC cells, their delivery via locoregional routes—specifically intra-arterial injection—had not been investigated, despite. ### In Vitro Transfection (N1-S1 Rat HCC Cells) | Parameter | Result | |---------------|------------| | Transfection efficiency (all PBAEs) | >50% GFP+ cells at various doses and w/w ratios | | Cell viability | >70% for.
CAR-T cell therapy has shown impressive success in hematological malignancies, but systemic toxicity (CRS, ICANS, on-target/off-tumor effects) and the complex, costly, individualized manufacturing process of autologous CAR-T cells hinder broader application. Universal allogeneic CAR-T cells have encountered safety concerns, with FDA halting some clinical trials. There is an urgent need for new strategies to overcome these barriers. --- - In vivo CAR-T induction with PBAE nanoparticles: Matthias Stephan's team achieved stable and transient expression of CD19-specific CAR in T cells via CAR-DNA and CAR-mRNA nanoparticles, respectively. Antitumor efficacy comparable to conventional lab-manufactured CAR-T cells without systemic toxicity. - Lentiviral in vivo CAR-T: Buchholz and colleagues induced in situ CAR-T cells in NSG mice with antitumor activity, but observed CRS and unexpected CAR-positive NK and NKT cells due to non-specific lentivi
Sérgio Scalzo, Anderson K. Santos, Heloisa A. S. Ferreira, Pedro A. Costa, Pedro H. D. M. Prazeres, Natalia J. A. Da Silva, Lays C. Guimarães, Mário De Morais E Silva, Marco T. R. Rodrigues Alves, Celso T. R. Viana, Itamar C. G. Jesus, Alice P. Rodrigues, Alexander Birbrair, Anderson O. Lobo, Freder
Cardiomyocytes are hard-to-transfect cells, and gene therapy for cardiovascular disease is limited by insufficient delivery to cardiac tissue, nucleic acid degradation, and safety concerns with viral vectors. A safe, effective non-viral platform for pDNA delivery to cardiomyocytes is needed. LNP4 was the top performer: ~1.3-fold higher GFP fluorescence than the second-best LNP8 and ~10-fold higher than the lowest performer LNP6. - LNP4 achieved >60% transfection efficiency at day 2 and >80% at day 4 in.
Sajid Iqbal, Alessandro Francisco Martins, Muhammad Sohail, Jingjing Zhao, Qi Deng, Muhan Li, Zhongxi Zhao
Polymeric gene delivery vectors require detailed physicochemical characterization to ensure reproducible formulation and efficient transfection. PBAE-447 is a promising biodegradable cationic polymer, but its simple nanoparticles can degrade or aggregate in aqueous/physiological conditions. This study characterizes PBAE-447 and evaluates PEGylation and lyophilization to improve stability and transfection. Polymer characterization: Mn 5,354; Mw 9,575; MP 4,934; PDI 1.7. Complete end-capping confirmed by ¹H NMR. - Solubility/buffering/swelling: PBAE-447 completely soluble at 10 µg/µL in NaAc; buffering capacity required.
Viral vectors dominate gene therapy but suffer from immunogenicity, insertional mutagenesis risk, cargo-size limits, and manufacturing challenges. Non-viral nanoparticles are safer, more economical, and easier to scale, but their clinical translation has been limited by inefficient delivery, especially for DNA and gene-editing cargoes that must reach the nucleus. There is a need to review non-viral nanomaterial design, delivery barriers, and. Viral vector dominance: Approximately 70% of gene therapy clinical trials to date have used viruses. - LNP gene editing in liver: A single administration of CRISPR-Cas9 LNPs achieved >97% reduction in target protein.
RNA-loaded lipid nanoparticles (LNPs) are clinically validated for siRNA and mRNA delivery, but their therapeutic performance depends critically on reproducible control of size, size distribution, zeta potential, and RNA encapsulation efficiency. Microfluidic devices offer precise mixing, rapid ethanol dilution, high reproducibility, high-throughput formulation screening, and continuous production, making them attractive for standardized LNP. Increasing PEG-lipid from 1 to 5 mol% reduced siRNA-LNP size from 54 nm to 28 nm; DLin-KC2-DMA LNPs achieved 50% FVII silencing at 0.01 mg/kg in mice. - Optimized mRNA-LNP formulations improved delivery and expression.
Therapeutic nucleic acids require safe and effective in vivo delivery vectors. Most gene and cell therapies rely on ex vivo gene delivery, which is laborious, time-consuming, and costly. PBAE is a promising biodegradable synthetic cationic polymer for in vivo gene delivery due to its transfection efficiency, biodegradability, and structural tunability. This review addresses the need for a systematic understanding of PBAE components and how. PBAE half-life: 1–7 h in aqueous conditions. - Freeze-thaw stability: PBAE NPs stored at −20 °C are stable through 8 freeze/thaw cycles without significant efficacy changes. - Optimal aqueous incubation: <10 min before.
Chemical Society Reviews* (as indicated by review format; exact volume/pages and DOI not included in the supplied file)2021ReviewNon-viral Gene Delivery
Yingshu Guo, Xiuping Cao, Xiaofei Zheng, Sk Jahir Abbas, Juan Li, Weihong Tan
Cancer treatment still relies heavily on chemotherapy, which causes severe damage to normal cells and systemic side effects. Safer, more selective drug delivery strategies are needed. Nanocarriers based on nucleic acids (NCNAs) are attractive because nucleic acids offer biocompatibility, low toxicity, programmable structures, controllable size, and modifiability—advantages that address limitations of cationic polymers, dendrimers, and inorganic nanoparticles. --- - DNA tetrahedron loaded with 5-fluorouracil: after 12 h co-incubation, the percentage of drug in cells was nearly 40%, demonstrating stable and efficient drug delivery. - DNA/RNA hybrid spherical nucleic acid with siRNA: rapidly absorbed by more than 60 kinds of cells; protected siRNA from enzymatic digestion and released it upon intracellular Dicer cleavage. - Se/Ru metal–organic nanoparticles + siRNA: enhanced cell uptake and promoted siRNA escape from endosomes/lysosomes, p
Ella S. Atsavapranee, Margaret M. Billingsley, Michael J. Mitchell
Adoptive T cell therapy has been revolutionized by genetic engineering, but viral transduction—the standard delivery method—has limitations including small cargo capacity, high immunogenicity, high manufacturing cost, and risks of genotoxicity/insertional mutagenesis. There is a need for improved non-viral delivery technologies to enable precise gene editing in T cells for cancer immunotherapy, especially for solid tumours and allogeneic applications. --- - Five FDA-approved CAR T cell therapies: Kymriah, Yescarta, Tecartus, Breyanzi, and Abecma. - Sleeping Beauty clinical trials: Patients with advanced NHL and ALL undergoing HSCT and CAR T cell infusion showed no acute or latent toxicities and no exacerbation of GVHD. - CRISPR/Cas9 first-in-human trial: T cells from three patients with refractory cancer were edited to remove endogenous TCRs and PD-1, achieving highly specific editing at targeted loci without clinical toxicity; edited T
Qurrat Ul Ain, Estefania V.R. Campos, Ariel Huynh, Dominik Witzigmann, Sarah Hedtrich
Gene therapies are powerful tools to prevent, treat, and cure human diseases, but their application to skin diseases has received little attention despite the easy accessibility of skin and urgent medical need. The main obstacle is the unique barrier properties of human skin, which severely limit absorption of biomacromolecules and efficient delivery of nucleic acid payloads. This review discusses current approaches, successes, and failures of cutaneous gene therapy and provides guidance for next-generation concepts, with delivery strategies as the major translational obstacle. --- - Skin barrier: Human skin efficiently absorbs only small molecules (MW < ~800 Da) with moderate lipophilicity (logP 1–3), making biomacromolecule delivery challenging. - Clinical trial landscape: Of 1052 gene therapy trials, only 23 focus on skin conditions; ~70% of ongoing gene therapy trials are viral vector-based. - LNP optimization: Zwitterionic lipid ce
Gene therapy requires safe, precise, efficient, and cost-effective delivery. Viral vectors are efficient but face manufacturing, cost, and safety challenges; non-viral vectors are safer and more scalable but often have low delivery efficiency. Magnetic nanoparticles (MNPs) have emerged as a promising strategy to enhance viral and non-viral gene delivery under an external magnetic field, while also enabling magnetic targeting, MRI tracking, and magnetic hyperthermia. --- - Magnetofection speed: MNP-based gene carriers can be attracted to cell surfaces in a few minutes, compared with hours for regular transfection. - Transfection enhancement: EMF enhanced lipid/SPIO-mediated transfection sixfold. PEI/Si@MNP with Dox and P-gp shRNA showed magnetically targeted delivery under 0.42 T for 12 h. - In vivo neuronal transfection: NeuroMag delivered EYFP-channelrhodopsin to rat visual cortex neurons with 72.66% and 86.63% expression at 3 and 30 d
Arun R. K. Kumar, Yufeng Shou, Brian Chan, Krishna L., Andy Tay
This review examines how viral vectors, electroporation, microfluidic systems, nanoparticles and high-aspect-ratio nanostructures can improve the efficiency, viability and scalability of transfecting primary immune cells for applications such as CAR-T, CAR-NK and gene editing. It highlights the trade-offs between delivery efficiency, cell fitness, cargo flexibility, manufacturing complexity and clinical translation.
Emily Harris, Devon Zimmerman, Eric Warga, Anil Bamezai, Jacob Elmer
Retroviral gene delivery for T cell therapies is expensive, semi-randomly integrating, and variable between patients. Non-viral alternatives are needed, but T cells are notoriously hard to transfect, especially primary T cells. Lipofectamine LTX was the best vehicle in Jurkat cells; optimized conditions in X-VIVO media yielded 63.0 ± 10.9% EGFP+ Jurkat cells vs. 23.1 ± 5.5% in RPMI. - Primary CD3+ T cells reached only 8.1 ± 0.8% EGFP+ under.
Tresa Lopez-Royo, Victor Sebastian, Laura Moreno-Martinez, Laura Uson, Cristina Yus, Teresa Alejo, Pilar Zaragoza, Rosario Osta, Manuel Arruebo, Raquel Manzano
CRISPR/Cas gene-editing components are often encoded on large plasmids (9–19 kb), which are difficult to encapsulate and transfect. Most PLGA nanoparticle (NP) gene-delivery studies use small nucleic acids (siRNA) or small plasmids (<6 kb), and the structural integrity and functional performance of large plasmids in PLGA NPs remain poorly characterized. There is a need to compare synthesis methods for encapsulating large plasmids without. pDNA stability: Batch ultrasound double emulsion completely degraded the 9.4 kb plasmid (standard) or left only 8.80% supercoiled (modified). Microfluidics-assisted double emulsion avoided degradation but produced only.
Rodgers, T., Muzzio, N., Watson, C., & Romero, G. (2021).
Gene-editing tools are large plasmid constructs that cannot spontaneously enter mammalian cells, and viral vectors are limited by cargo size, immunogenicity, and manufacturing challenges. PBAE nanoparticles are promising nonviral carriers, but unmodified PEG-PDHA nanoparticles release encapsulated plasmid DNA too rapidly for efficient intracellular delivery of large gene-editing cargo such as piggyBac transposon. Encapsulation and release: Optimal polymer/plasmid molar ratio = 0.36. Unmodified PEG-PDHA NPs released essentially all PBCAG within 2 h. Layer-by-layer NPs showed no detectable release within 24 h but transfection was.
Nanomedicine translation is limited by non-scalable, batch-variable bulk NP formulation methods that often produce large, polydisperse particles. Microfluidics offers precise control over microscale mixing and nanoprecipitation, enabling more reproducible NPs with controlled size, size distribution, and loading. This review summarizes microfluidic advances for lipid-, polymer-, and inorganic-based NPs. Representative quantitative findings from reviewed literature: - SHM-produced LNPs: 60–90 nm vs ~180 nm by pipette mixing; >90% hepatic gene silencing; up to 7-fold increased mRNA potency. - Parallelized SHM: 72 mL/min;.
Cancer immunotherapy has shown efficacy but is limited by serious adverse effects, nonspecific inflammation, and challenges in spatiotemporal control of immune responses. Nanomedicine and functionalized nanosystems offer programmable pharmacokinetics, co-delivery of immunomodulators, and targeted delivery, potentially improving immunotherapy outcomes. Antigen-capturing NPs (AC-NPs) plus αPD-1 improved cure rate by 20% in B16F10 melanoma, expanded CD8⁺ cytotoxic T cells, and increased CD4⁺/Treg and CD8⁺/Treg ratios. - Cationic lipid-assisted PEG-b-PLGA NPs (CLANs).
Cervical cancer is immunologically relevant because retained high-risk HPV antigens can be recognized as foreign, making immunotherapy attractive. However, conventional immunotherapy is limited by variable efficacy, immune evasion, cytokine toxicity, and poor targeting. Nanomedicine may improve delivery, targeting, pharmacokinetics, and safety of immunotherapeutic agents. Pembrolizumab showed increased activity in cervical cancer in KEYNOTE-158 and promising antitumor activity in phase Ib KEYNOTE-028. - Nivolumab in phase II NRG-GY002: minimal response rate, median survival 14.5 months,.
Nucleic acid vaccines (DNA and mRNA) offer rapid design, adaptability to changing pathogen strains, and stimulation of both humoral and cellular immunity, but their clinical translation is limited by inefficient delivery—nucleic acids are rapidly degraded by endogenous nucleases, have poor cellular uptake due to their negative charge, and must cross cellular barriers (cytoplasm for mRNA, nucleus for DNA). Nanoparticles (NPs) are promising. mRNA-1273 (Moderna): phase III trial with 30,000+ participants showed 95% efficacy in preventing COVID-19; phase I showed dose-dependent antibody responses (day 57 GMT: 299,751, 782,719, and 1,192,154 for 25, 100, and.
Rumiana Tenchov, Robert E. Bird, Allison E. Curtze, Qiongqiong Angela Zhou
Lipid nanoparticles (LNPs) have become key delivery vehicles across pharmaceuticals, with renewed prominence as the delivery platform for COVID-19 mRNA vaccines. The field lacks a broad, quantitative landscape of LNP research across liposomes, solid lipid nanoparticles, nanostructured lipid carriers, cationic lipid–nucleic acid complexes, and newer architectures. This review provides an overview of LNP structures, properties, applications, and. LNP research is dominated by pharmaceutical applications. The fastest-growing research areas are pharmaceuticals, food and feed, and cosmetics. - The term “liposome” appears in ~147,000 documents from 2000–2020, versus.
Polymeric nanoparticles are widely used for cancer theranostics because of their biocompatibility, biodegradability, and structural versatility, but conventional delivery still faces poor target specificity, uncontrolled release, and systemic toxicity. Stimuli-responsive polymeric nanoplatforms are proposed to exploit tumor microenvironment (TME) and external triggers to enable precise, on-demand drug/gene release at tumor sites. Representative findings highlighted in the review: - TME gradients: tumor extracellular pH is approximately 5.7–6.9 versus blood pH 7.4; intracellular GSH is 2–10 mM versus extracellular 2–10 µM; tumor ROS is about.
CRISPR/Cas9 genome editing holds major therapeutic promise, but in vivo delivery is arguably the most difficult barrier to clinical translation. Efficient, selective transport of editing components to target cells/tissues while minimizing off-target effects remains unresolved. This review critiques lipid- and polymer-based nanoparticle delivery strategies for CRISPR/Cas9. LPO1 lipid nanoparticle: >97% knockdown of mouse TTR protein. - BAMEA-O16B bioreducible LNP: up to 90% GFP knockout in HEK cells; 80% PCSK9 knockdown in mouse serum after IV injection. - Cationic lipid NP with.
Anne Vejux, Dehbia Abed-Vieillard, Khadija Hajji, Amira Zarrouk, John J. Mackrill, Shubhrima Ghosh, Thomas Nury, Aline Yamminse, Mohamed Zaibih, Wafa Mihoubi, Habiba Bouchabi, Boubker Nasser, Yael Grosjean, Gérard Lizard
7-Ketocholesterol (7KC) and 7β-hydroxycholesterol (7β-OHC) are major auto-oxidation products of cholesterol that accumulate in cardiovascular disease, age-related macular degeneration (ARMD), neurodegenerative disorders, and inflammatory bowel diseases. While their cytotoxicity (oxidative stress, organelle dysfunction, cell death, inflammation) is well documented in cell culture, the field lacks a consolidated understanding of how these oxysterols behave in vivo and which model systems (in vitro, animal, and emerging alternatives) are most appropriate for identifying pharmacological targets and protective molecules. This review addresses that gap by systematically surveying the models and mechanisms reported to date. --- 1. Oxiappotophagy is the dominant cell death mode: 7KC and 7β-OHC simultaneously induce oxidative stress (ROS overproduction: O₂⁻, H₂O₂), apoptosis (caspase activation, PARP cleavage, DNA fragmentation), and autophagy (
Melanoma is a poorly immunogenic and highly aggressive skin cancer with limited long-term survival for advanced disease. Dendritic cell (DC)-based vaccines are promising immunotherapies, but their efficacy relies on critical factors including DC maturation state and efficient antigen delivery. Nanoparticulate delivery systems can enhance antigen delivery to ex vivo-generated DCs, mediate DC maturation (adjuvanticity), and promote cytoplasmic antigen presentation through MHC class I—potentially leading to potent antigen-specific immune responses. This review addresses the need to consolidate and evaluate the role of different nanoparticulate delivery systems in ex vivo-generated DC-based vaccines against melanoma. --- - Fusogenic liposomes (FLs): TCL/FLs-pulsed DCs significantly inhibited tumor growth until 17 days post-inoculation; superior to TCL/CLs-pulsed DCs. - Cationic liposomes (Srinivas et al.): Lipid 5 (shikimoyl headgroup) indu
