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.
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.
mRNA therapeutics have fueled hope to combat incurable diseases, but insufficient understanding of mRNA instability, immunogenicity, and delivery has impeded progress. The review argues that mRNA-based drugs—especially after the COVID-19 vaccine success—can become powerful, versatile tools, and that mRNA optimization and delivery systems are the key bottlenecks to solve. COVID-19 mRNA vaccines: ~90% effectiveness for full vaccination, 80% for partial; BNT162b2 95% efficacy in phase III; mRNA-1273 94.1% efficacy; Omicron-neutralizing antibodies largely undetectable in most recipients,.
Immunotherapy has improved outcomes in cancer and infectious disease, but off-target effects, systemic toxicities, and variable efficacy remain limiting. Nanoscale engineering offers ways to manipulate immune cell functions—enhancing immunity against cancers and pathogens, controlling the site of immune response, and promoting tolerance—by tuning nanoparticle size, shape, charge, and surface chemistry. CL4H6 lipid nanoparticles silenced STAT3 and HIF-1α in tumor-associated macrophages by 37% and 48%, respectively, increased M1 markers, reduced tumor size, and altered cytokine profiles in a B16-F10 model. -.
Randall A. Meyer, Sarah Y. Neshat, Jordan J. Green, Jose Luis Santos, And Anthony D. Tuesca
mRNA gene therapy has broad therapeutic potential (protein replacement, vaccines, regenerative medicine, oncology), but clinical translation is limited by inadequate understanding of how to target specific organs or cell types for protein expression. Reports on material properties and administration routes are disparate, preventing a global understanding of how these factors contribute to organ targeting for mRNA delivery. Liver targeting: LNPs intrinsically target liver via ApoE-mediated uptake through LDLR. Optimized ionizable lipids increased liver luciferase expression 18-fold. Oxidized cholesterol variants improved protein 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.
Johan Karlsson, Stephanie Y. Tzeng, Shayan Hemmati, Kathryn M. Luly, Olivia Choi, Yuan Rui, David R. Wilson, Kristen L. Kozielski, Alfredo Quiñones-Hinojosa, Jordan J. Green
Systemic delivery of RNA therapeutics with polymer-based nanocarriers is limited by poor colloidal stability in blood and inefficient intracellular delivery of siRNA to the cytosol. There is a need for nanoparticles that remain stable extracellularly but rapidly release RNA intracellularly. Crosslinking: Molecular weight increased by 42.1% (Mn) and 27.7% (Mw); acrylate peak intensity decreased by 79.1% ± 0.3%. - Serum stability: XbNPs retained siRNA encapsulation in 50% serum; non-crosslinked formulations.
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.
Est-Witte Se, Livingston Nk, Omotoso Mo, Green Jj, Schneck Jp.
T cell therapies (TIL, TCR, CAR T) show clinical efficacy but are limited by high cost, lengthy ex vivo manufacturing, patient variability, and poor persistence. Nanoparticles offer a modular, universal platform to improve T cell therapy at every stage—from antigen presentation and T cell activation to T cell maintenance—while reducing cost and broadening patient accessibility. Nano-aAPC E+E expanded Kb-TRP2 cognate cells from ~0.03% to 17.6% by day 7; nano-aAPCs expanded MART-1-specific CD8⁺ T cells more effectively than autologous DCs and CD3/CD28 Dynabeads, with higher stem cell memory.
mRNA vaccines are promising but naked mRNA is fragile, susceptible to enzymatic degradation, poorly taken up by cells due to electrostatic repulsion, and can trigger innate immune responses. Self-assembly offers a versatile approach to prepare delivery vehicles with customizable properties. The review discusses design and self-assembly of mRNA vaccines, materials commonly used, physicochemical characteristics, routes of administration, and. BNT162b2: 95.0% efficacy in phase III (8 COVID-19 cases in vaccine group vs 162 in placebo); 94.6% efficacy including prior infection; ~52% efficacy between first and second dose. - mRNA-1273: 94.1% efficacy in phase.
Kelly R. Rhodes, Randall A. Meyer, Justin Wang, Stephany Y. Tzeng, Jordan J. Green
Regulatory T cell (Treg)-based therapeutics show promise for treating autoimmune diseases and preventing transplant rejection, but adoptive Treg transfer is expensive, complex, and difficult to implement. "Off-the-shelf" non-cellular alternatives that can induce endogenous Tregs in vivo are needed. Existing artificial antigen presenting cells (aAPCs) are typically used for ex vivo T cell expansion and have limited application for immune. ### Protein Conjugation & aAPC Characterization | Parameter | PLGA aAPC | PLGA/PBAE aAPC | Significance | |---------------|---------------|--------------------|------------------| | Anti-CD3 conjugation (1× dose) |.
N.N. Parayath, S.B. Stephan, A.L. Koehne, P.S. Nelson, M.T. Stephan
CAR T and TCR T cell therapies are powerful but face prohibitive costs and manufacturing complexity due to the need for leukapheresis, ex vivo transduction, expansion, and quality control—all under GMP conditions. An "off-the-shelf" injectable nanocarrier that can transiently reprogram circulating T cells in vivo would bypass these barriers, enabling scalable, cost-effective, and repeatable dosing for a wide range of diseases including cancer. ### NP Characterization & Ex Vivo Transfection | Parameter | Result | |---------------|------------| | NP size (PBAE/mRNA polyplex) | ~100 nm | | mRNA encapsulation efficiency | >90% | | T cell transfection efficiency |.
John Choi, Yuan Rui, Jayoung Kim, Noah Gorelick, David R. Wilson, Kristen Kozielski, Antonella Mangraviti, Eric Sankey, Henry Brem, Betty Tyler, Jordan J. Green, Eric M. Jackson
Medulloblastoma (MB) and atypical teratoid/rhabdoid tumor (AT/RT) are common pediatric brain malignancies. Radiation carries high risks of developmental sequelae in young children, and current treatments remain inadequate. Viral gene therapy has safety concerns in pediatric patients, so a safe, effective nonviral polymeric nanoparticle system is needed for gene delivery to pediatric CNS tumors. Transfection: Several PBAE formulations achieved >50% transfection in both BT-12 and D425 cells with low cytotoxicity. Optimized formulations significantly outperformed 25 kDa PEI and Lipofectamine 3000. - Uptake: 447.
ACS Biomaterials Science & Engineering (as indicated in the supplied file header; exact volume/pages and DOI were not in2020ResearchNon-viral Gene Delivery
Denis Routkevitch, Deepti Sudhakar, Marranne Conge, Mahita Varanasi, Stephany Y. Tzeng, David R. Wilson, Jordan J. Green
The mechanism by which cationic polymers with titratable amines mediate endosomal escape and cytosolic delivery of nucleic acids remains poorly understood. Buffering capacity alone has often failed to predict transfection efficacy, so the study examines whether the effective pKa of poly(beta-amino ester)s (PBAEs) governs cytosolic delivery and transfection. Transfection efficacy: PBAEs achieved up to 99% in HEK293T, 65% in B16-F10, and 90% in GB319 cells, with high viability. - Effective pKa: PBAE 746 = 6.16, PBAE 446 = 6.95, PBAE 447 = 7.15. PEI showed broad buffering.
ACS Biomaterials Science & Engineering (as indicated in the file header; exact volume/pages and DOI were not included in2020ResearchNon-viral Gene Delivery
Denis Routkevitch, Deepti Sudhakar, Marranne Conge, Mahita Varanasi, Stephany Y. Tzeng, David R. Wilson, Jordan J. Green
The mechanism by which cationic polymers with titratable amines mediate endosomal escape and cytosolic delivery of nucleic acids remains poorly understood. Buffering capacity alone has often failed to predict transfection efficacy, so this study examines whether the effective pKa of poly(beta-amino ester)s (PBAEs) governs cytosolic delivery and transfection. Transfection efficacy: PBAEs achieved up to 99% in HEK293T, 65% in B16-F10, and 90% in GB319 cells, with high viability. - Effective pKa: PBAE 746 = 6.16, PBAE 446 = 6.95, PBAE 447 = 7.15. PEI showed broad buffering.
Rui Yan Yuan, Mahita Varanasi, Shanelle Mendes, Hannah M. Yamagata, David R. Wilson, Jordan J. Green
CRISPR-Cas9 gene editing requires efficient intracellular delivery of large cargo. Viral vectors have packaging and production limitations, and nonviral CRISPR plasmid delivery had mainly been studied for single-site knockout, not for gene deletion requiring two cleavage events. The transfection requirements for 1-cut vs. 2-cut edits were unclear. Transfection: >80% of cells were transfected in both HEK293T and B16-F10 using GFP reporter. However, geometric mean expression in HEK293T was nearly 1 order of magnitude higher than B16-F10. - 1-cut knockout: Up to 70%.
Karlsson, J.; Rhodes, K. R.; Green, J. J.; Tzeng, S. Y
Non-viral gene delivery needs safe and efficient carriers. PBAEs are biodegradable cationic polymers with promising transfection activity, but they face challenges such as instability in physiological fluids, positive-charge toxicity, and limited systemic delivery. This review summarizes PBAE properties, advantages, limitations, and strategies to customize them for diverse gene delivery applications. PBAE/DNA nanoparticles functionalized with T-cell targeting and nuclear localization signal achieved >80% transfection of T cells. - Bioreducible PBAEs achieved near-complete siRNA knockdown in patient-derived.
Wahane A, Waghmode A, Kapphahn A, Dhur K, Gupta A, Bahal R
Nucleic acid analogs and genome-editing proteins have broad therapeutic potential, but clinical translation is limited by poor cell- and organ-specific delivery, endosomal entrapment, enzymatic degradation, and short bioavailability. The review examines lipid- and polymer-based non-viral vectors as strategies to overcome these barriers. DOTMA liposomes: ~100% plasmid DNA entrapment and 5–100-fold higher transfection than calcium phosphate or DEAE-dextran. - SPLPs: ~70 nm particles with ~70% encapsulation; 100–1,000-fold gene expression in distal tumor.
Wilson, D. R., Rui, Y., Siddiq, K., Routkevitch, D., & Green, J. J. (2019).
Nonviral gene delivery remains limited by low transfection efficacy, especially under physiological serum conditions and at low nanoparticle doses. Although linear poly(β-amino ester)s (PBAEs) are effective, branching structure had not been systematically evaluated in well-defined PBAE-like poly(ester amine) systems. Transfection efficacy: BEAQs achieved up to 99% transfection in HEK293T and 77% in ARPE-19 cells in 10% serum, exceeding linear PBAE and commercial reagents. - High serum: Under 50% serum, B8-50% transfected 98% of.
Yingqi Xu, Dingxin Liu, Jie Hu, Peirong Ding, Meiwan Chen
Triple-negative breast cancer (TNBC) lacks effective targeted therapy. Nonviral co-delivery of an anticancer drug and a therapeutic gene may improve treatment, but efficient tumor-targeted co-delivery remains challenging. TRAIL has shown limited clinical benefit alone, and embelin (EMB) can inhibit XIAP to promote apoptosis. Nanoparticle properties: PPNP CMC = 0.0065 mg/mL. HA coating reduced hemolysis from 37.72% to <0.5%. HA/PPNP remained stable in 10% FBS for 24 h, while PPNP aggregated to ~4000 nm. - Uptake: HA/PPNP increased Nile red.
Samuel A. Smith, Laura I. Selby, Angus P. R. Johnston, And Georgina K. Such
Many emerging therapies require delivery of biological cargo into the cytosol, but nanoparticle delivery systems are hindered by entrapment and degradation in the endo/lysosomal pathway. Endosomal escape remains a rate-determining step for non-viral delivery. The review highlights the need for greater understanding of escape mechanisms and improved quantification methods to enable rational design of safer, more efficient polymeric delivery. Escape efficiency is low: Zerial and co-workers demonstrated that <2% of siRNA delivered using lipid nanoparticles is delivered to the cytosol. - Calcein assay: 27 kDa PDEAEMA nanoparticles showed leakage of calcein.
Thomas, T. J.; Tajmir-Riahi, H.-A.; Pillai, C. K. S
Gene therapy requires efficient delivery of genetic material, but viral vectors carry risks of immunogenicity, toxicity, and cancer formation. Cationic polymers such as polyethyleneimine (PEI) can condense DNA and facilitate delivery, but their lack of biodegradation causes significant toxicity due to polymer accumulation in tissues. This review addresses the need for biodegradable polymeric gene delivery vehicles developed over the past decade. PEI: ~20% of nitrogen protonated at physiological pH; 22 kDa linear PEI showed more facile nuclear delivery than branched PEI. Non-degradable PEI causes severe cytotoxicity. - Biodegradable PEI derivatives: Reducible.
Cordeiro, R. A.; Serra, A.; Coelho, J. F. J.; Faneca, H
Gene therapy requires safe and effective delivery vectors. PBAEs are biodegradable cationic polymers with excellent gene delivery properties, but a comprehensive and critical overview of their use as gene carriers is lacking. This review provides the first comprehensive and critical overview of PBAE-based gene delivery systems, from discovery to therapeutic applications, with emphasis on rational design and structure–performance relationships. First library screening (2001): 140 PBAEs from 7 diacrylates and 20 amines; C93 and G28 gave 4–8 times higher transfection than PEI. - Large library (2003): 2,350 structurally unique PBAEs synthesized in a single day;.
Elana Ben-Akiva, Kelly R. Rhodes, Randall A. Meyer, Jordan J. Green
Artificial antigen-presenting cells (aAPCs) are promising for immune modulation, and anisotropic (ellipsoidal) particles have shown improved T-cell stimulation over spherical particles. However, widely used methods for generating anisotropic particles, such as thin-film stretching, can be difficult to implement reproducibly. A standardized, scalable protocol is needed for fabricating biodegradable anisotropic aAPCs with tunable size, shape, and. Particle size: Spherical nanoparticles = 237.3 ± 4.0 nm by DLS and 224 nm by NTA; spherical microparticles = 3 ± 1 µm. - Aspect ratios: Spherical = 1.05 ± 0.04; 1D-stretched prolate ellipsoidal = 3.6 ± 0.8; 2D-stretched.
