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.
Yue Wang, Shi-Kun Zhou, Yan Wang, Zi-Dong Lu, Yue Zhang, Cong-Fei Xu, Jun Wang
PD-1/PD-L1 blockade therapy is successful but often yields poor benefits due to insufficient T-cell infiltration and low intratumoral concentrations of PD-1/PD-L1 inhibitors. While strategies exist to increase either T-cell recruitment or inhibitor delivery, none actively recruit T cells while achieving tumor-specific delivery of PD-L1 inhibitors to specifically eliminate inhibition of tumor-infiltrating T cells. A strategy that addresses both. ### Nanoparticle Characterization | Parameter | Value | |---------------|-----------| | Hydrodynamic diameter | 107.2 nm | | PDI | ~0.2 | | Zeta potential | +15.3 mV | | Stability in 10% FBS | ≥5 days (size/PDI stable).
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 is FDA/EMA-approved for B cell malignancies and multiple myeloma, but manufacturing relies on viral vectors—associated with safety concerns, high cost, and production challenges—or electroporation, which can be highly cytotoxic. Nanosystems may offer a safer, cost-effective alternative, but T cells are difficult to transfect, so rational design of lipid- and polymer-based carriers is urgently needed. Si-PDMAEMA-pDNA achieved 46% transfection in Jurkat cells and 44% in primary human T cells; pDNA/PEI-based systems reached 51% in Jurkat and 60% in primary human T cells. - pIAE + anti-CD3 + PiggyBac transposon.
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
Hamideh Parhiz, Jacob S. Brenner, Priyal N. Patel, Tyler E. Papp, Hamna Shahnawaz, Qin Li, Ruiqi Shi, Marco E. Zamora, Amir Yadegari, Oscar A. Marcos-Contreras, Ambika Natesan, Norbert Pardi, Vladimir V. Shuvaev, Raisa Kiseleva, Jacob W. Myerson, Thomas Uhler, Rachel S. Riley, Xuexiang Han, Michael
Nucleoside-modified mRNA lipid nanoparticle (modmRNA-LNP) technology is clinically successful, but its behavior in pre-existing inflammatory or immune-challenged conditions is poorly characterized. This gap raises the risk of adverse effects when modmRNA-LNPs are administered to patients with ongoing inflammation. IE with LPS + modmRNA-LNP: Serum IL-6 increased 14-fold vs LPS alone and 94-fold vs modmRNA-LNP in naive mice. Liver MIP-2 increased 11-fold vs LPS alone and 52-fold vs modmRNA-LNP in naive mice. Liver MIP-2 mRNA.
Kelsey L. Swingle, Margaret M. Billingsley, Sourav K. Bose, Brandon White, Rohan Palanki, Apeksha Dave, Savan K. Patel, Ningqiang Gong, Alex G. Hamilton, Mohamad-Gabriel Alameh, Drew Weissman, William H. Peranteau, Michael J. Mitchell
Congenital protein deficiencies often begin irreversible pathology in utero, but postnatal treatments are limited. In utero mRNA delivery could enable protein replacement before disease onset, yet LNP stability in fetal amniotic fluid and how it affects mRNA delivery had not been previously investigated. LNP A12 was most stable in mouse amniotic fluid; A1 was least stable. A12 had significantly higher bound protein content than A1 (p < 0.0021). - Percent change in PDI inversely correlated with in vitro luciferase.
Martijn J.W. Evers, Wenjuan Du, Qiangbing Yang, Sander A.A. Kooijmans, Aryan Vink, Mies Van Steenbergen, Pieter Vader, Saskia C.A. De Jager, Sabine A. Fuchs, Enrico Mastrobattista, Joost P.G. Sluijter, Zhiyong Lei, Raymond Schiffelers
Modified mRNA (modRNA) is a promising cardiac regeneration therapeutic, but naked mRNA is large, negatively charged, and rapidly degraded. It is unknown whether systemically administered lipid nanoparticles (LNPs) can functionally deliver mRNA to ischemic/damaged myocardium after myocardial infarction. LNPs were <100 nm with PDI <0.2 and 95–99% mRNA encapsulation. - After ischemia-reperfusion, fluorescent LNPs accumulated in the infarcted area of the heart at 4 h and 24 h, but not in sham or control hearts. -.
Yihua Pei, Yanjie Bao, Cristiano Sacchetti, Juthamart Brady, Kyra Gillard, Hailong Yu, Scott Roberts, Kumar Rajappan, Steven P. Tanis, Carlos G. Perez-Garcia, Padmanabh Chivukula, Priya P. Karmali
Systemic LNP delivery predominantly targets the liver, while extrahepatic delivery—especially to lung airway epithelium for diseases such as cystic fibrosis—remains challenging. DOTAP is a common cationic lipid for lung gene delivery but is racemic, pseudo-glyceryl, and slowly biodegradable. Novel readily hydrolysable DOTAP analogues are needed for safer and more effective inhaled/airway mRNA delivery. DOTAP+ and L1–L4 LNPs: <100 nm, >95% mRNA encapsulation, high mRNA purity; DOTAP− had ~71% encapsulation. - Freeze-thaw: all formulations retained size, PDI, and encapsulation within ~10% of initial values. -.
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.
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.
Farough Marofi, Roza Motavalli, Vladimir A. Safonov, Lakshmi Thangavelu, Alexei Valerievich Yumashev, Markov Alexander, Navid Shomali, Max Stanley Chartrand, Yashwant Pathak, Mostafa Jarahian, Sepideh Izadi, Ali Hassanzadeh, Naghmeh Shirafkan, Safa Tahmasebi, Farhad Motavalli Khiavi
CAR T cell therapy has transformed treatment of hematologic malignancies, but its efficacy in solid tumors remains unsupported. Solid tumors present distinct barriers—antigen heterogeneity, poor T cell trafficking/infiltration, and an immunosuppressive tumor microenvironment (TME)—that limit CAR T cell function. This review addresses these barriers and evaluates emerging strategies to improve CAR T cell therapy in non-hematologic malignancies. --- - Ovarian cancer: MSLN-CAR NK cells significantly killed MSLN⁺ ovarian cancer cells (SK-OV-3, OVCAR-3) *in vitro*. MUC16-specific CAR T cells eradicated malignant cells in mouse models. TAG72-CAR T and FRα-CAR T inhibited ovarian cancer growth. - Breast cancer: MUC28z CAR T cells (targeting tMUC1) reduced TNBC tumor proliferation and survival in a xenograft model. HRG1β-based CAR T cells inhibited breast cancer via HER family receptors. Anti-HER2 CAR T cells triggered cell death in HER2-overex
CAR-T cell therapy has produced remarkable clinical responses in certain B cell leukemias and lymphomas, but major limitations remain—including life-threatening toxicities, limited efficacy in solid tumors, antigen escape, poor persistence/trafficking, and an immunosuppressive microenvironment. This review addresses these barriers and discusses recent innovations in CAR-T engineering to improve efficacy and safety in both hematological malignancies and solid tumors. --- - Antigen escape: 70–90% of relapsed/refractory ALL patients show durable responses to CD19 CAR-T, but 30–70% of recurrent disease involves CD19 downregulation/loss. BCMA loss also observed in multiple myeloma. - Dual targeting: CD19/CD22 and CD19/BCMA dual-targeted CAR-T cells show promising efficacy and favorable safety in early clinical trials. Tandem HER2/IL13Rα2 CARs improved anti-tumor activity and decreased antigen escape in glioblastoma models. - Toxicity reducti
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
Ian C. Miller, Ali Zamat, Lee-Kai Sun, Hathaichanok Phuengkham, Adrian M. Harris, Lena Gamboa, Jason Yang, John P. Murad, Saul J. Priceman, Gabriel A. Kwong
CAR T cell therapy for solid malignancies typically results in poor responses. Systemic administration of immunomodulatory biologics (cytokines, BiTEs) can augment T cell activity but off-target toxicity narrows the therapeutic window. A method to spatially and temporally control transgene expression by engineered T cells at tumor sites—without systemic exposure—could improve safety and efficacy. ### Thermal Switch Engineering | Parameter | Result | |---------------|------------| | Best HSE repeat number | 7 repeats (5H-7H best; 2H-4H lower response) | | Best core promoter | YB (synthetic) — ~60-fold induction.
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.
Sarah J. Shepherd, Claude C. Warzecha, Sagar Yadavali, Rakan El-Mayta, Mohamad-Gabriel Alameh, Lili Wang, Drew Weissman, James M. Wilson, David Issadore, Michael J. Mitchell
Microfluidic mixing can produce precise LNPs but is limited in throughput, while bulk mixing is scalable but yields larger, heterogeneous particles with variable potency. A scalable microfluidic device is needed to produce potent RNA-LNPs across discovery and clinical scales. PMD achieved 18.4 L/h production, >100-fold higher throughput than a single microfluidic channel. - Mixing performance was uniform across 10× and 128× devices; 90% mixing channel length correlated linearly with ln(Pe).
siRNA therapeutics are limited by poor cellular uptake, serum nuclease degradation, and inefficient endosomal/lysosomal escape. Chitosan is biocompatible and biodegradable but has low transfection efficiency, largely due to poor buffering capacity and weak endosomal escape. Histidine grafting was explored to add imidazole groups (pKa ~6) that enhance proton-sponge buffering and siRNA delivery. Synthesis: FT-IR showed amide C=O at 1640 cm⁻¹ and N–H bending shift from 1590 to 1522 cm⁻¹; histidine O–H at 3016 cm⁻¹ disappeared. XRD showed loss of histidine crystal peaks at 18.8° and 24.3°, and HGCS polymers were.
Jiayu Zhou, Yishu Zhai, Jumei Xu, Tian Zhou, Lian Cen
PLGA microspheres are widely used for controlled drug release, but they often exhibit pronounced initial or mid-term burst release. A strategy is needed to finely tune drug release kinetics and suppress burst release, especially for water-soluble drugs. MSNs: Average size 119 nm; specific surface area 902.53 m²/g; pore volume 1.15 cm³/g; mean pore diameter 5.09 nm; maximum RB loading ~110 mg/g. - MSN-RB release: ~95% cumulative release within 56 h; Korsmeyer–Peppas n =.
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 (
Annika M. Frank, Angela H. Braun, Lea Scheib, Shivani Agarwal, Irene C. Schneider, Floriane Fusil, Severine Perian, Ugur Sahin, Frederic B. Thalheimer, Els Verhoeyen, Christian J. Buchholz
Conventional lentiviral vectors (LVs) are not selective for T cells and cannot efficiently transduce resting or minimally stimulated T lymphocytes, which limits their use for direct in vivo T-cell gene delivery. There is a need for vectors that can both target T cells specifically and activate them sufficiently to permit gene transfer without prior ex vivo activation. --- - Selectivity: CD3-LVs transduced CD3(^+) Jurkat cells but not CD3(^-) Molt4.8 cells. VSV-LV transduced HUVEC (86.74%), HepG2 (32.53%), and Nalm-6 (97.50%) cells, whereas CD3-LVs showed <0.3% transduction in these off-target cells. - Nonactivated T cells: CD3-LVs transduced freshly isolated PBMCs in the absence of cytokines or with IL-2 or IL-7/IL-15 at efficiencies similar to peak activation. VSV-LV was substantially less efficient under these conditions. - Whole blood: CD3-LVs mediated T-cell gene transfer in whole blood without additional stimuli. VCNs: TR66-LV 1.17
Bispecific T-cell engager (BiTE) immunotherapy is a promising cancer treatment, but clinical application is limited by high production costs and short in vivo half-life of BiTE proteins. A gene delivery system that enables sustained in vivo expression of BiTE could overcome these limitations, but existing non-viral vectors have low in vivo transfection efficiency. Metal-organic frameworks (MOFs) have been explored for nucleic acid delivery but. ### MOF Characterization & DNA Loading | Parameter | Result | |---------------|------------| | MOF size (TEM) | ~300 nm length × ~100 nm width | | MOF size (DLS) | 187.8 nm | | Zeta potential (MOF) | +24.7 mV | | Zeta.
John C. Charpentier, Di Chen, Philip E. Lapinski, Jackson Turner, Irina Grigorova, Joel A. Swanson, Philip D. King
Macropinocytosis has been shown to regulate the growth of Ras-transformed tumor cells under amino acid (AA)-limiting conditions by providing extracellular proteins that are degraded into AA to fuel cell growth and activate mTORC1. However, whether macropinocytosis regulates the growth of non-transformed mammalian cells, such as primary T lymphocytes, was unknown. Understanding whether macropinocytosis is required for T cell growth has. ### T Cell Macropinocytosis (Probe Uptake) | Cell Type | Probe | Stimulation | Fold Increase (Stimulated vs. Unstimulated) |.
Julie J.-J. Tang, Alina P. Sung, Mariel J. Guglielmo, Laura N.-G., Doug Redelman, James Smith-Gagen, Diane Hudig
Monoclonal antibody anti-tumor therapies rely on NK cell-mediated antibody-dependent cell-mediated cytotoxicity (ADCC), but patient responses vary widely. While tumor antigen escape is one cause of treatment failure, the role of inter-individual variation in NK cell serial killing capacity—the ability of a single NK cell to kill multiple target cells sequentially—has received little attention. A predictive biomarker for serial ADCC capacity. ### Killing Frequency Variability (E:T=1:4, n=24 donors) | Parameter | All Donors | Females | Males | |---------------|----------------|-------------|-----------| | KF range | 1.1 - 2.2 | 1.1 - 2.2 | 1.2 - 2.1 | | Mean.
