bims-novged 2022-07-24 papers

bims-novged

Biomed News

on Non-viral vectors for gene delivery

Issue of 2022‒07‒24
twelve papers selected by
the Merkel lab
Ludwig-Maximilians University

Effect of PEG anchor in PEGylation of folate-modified cationic liposomes with PEG-derivatives on systemic siRNA delivery into the tumor.
Transferrin receptor targeting segment T7 containing peptide gene delivery vectors for efficient transfection of brain tumor cells.
The Application of Sensitive Nano-Confined Nanoparticle System Using siRNA Targeting Extracellular Signal-Regulated Kinase (ERK)/Mitogen-Activated Protein Kinase (MAPK) Pathway in Gastrointestinal Cancer.
pH-sensitive packaging of cationic particles by an anionic block copolymer shell.
Nonclinical safety evaluation of a novel ionizable lipid for mRNA delivery.
Hyaluronic Acid-Modified Nanoplatforms as a Vector for Targeted Delivery of Autophagy-Related Gene to the Endometriotic Lesions in Mice.
Redox-Unlockable Nanoparticle-Based MST1 Delivery System to Attenuate Hepatic Steatosis via the AMPK/SREBP-1c Signaling Axis.
Polycation Architecture Affects Complexation and Delivery of Short Antisense Oligonucleotides: Micelleplexes Outperform Polyplexes.
Thiol-Disulfide Exchange as a Route for Endosomal Escape of Polymeric Nanoparticles.
Polymeric nanoparticles for dual-targeted theranostic gene delivery to hepatocellular carcinoma.
The effect of drug loading and multiple administration on the protein corona formation and brain delivery property of PEG-PLA nanoparticles.
Cationic nanoparticles enhance T cell tumor infiltration and antitumor immune responses to a melanoma vaccine.

J Drug Target. 2022 Jul 21. 1-21

Effect of PEG anchor in PEGylation of folate-modified cationic liposomes with PEG-derivatives on systemic siRNA delivery into the tumor.

Min Tang, Sho Sakasai, Hiraku Onishi, Kumi Kawano, Yoshiyuki Hattori.

  In this study, we prepared small interfering RNA (siRNA)/cationic liposome complexes (lipoplexes) modified with folate (FA)-polyethylene glycol (PEG, MW 2000, 3400 or 5000)-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) to facilitate their uptake into tumor cells via folate receptor (FR), and with PEG1600-cholesterol (PEG1600-Chol) or PEG2000-chondroitin sulfate conjugate (PEG2000-CS), to enhance their systemic stability. Among the FA-PEG-modified siRNA lipoplexes, 0.5 mol% FA-PEG5000-DSPE-modified lipoplexes with 2.5 mol% PEG2000-CS or PEG1600-Chol (LP-0.5F5/2.5P2-CS and LP-0.5F5/2.5P1.6-CL, respectively) exhibited selective growth inhibition of human nasopharyngeal carcinoma KB cells through transduction with polo-like kinase 1 (PLK1) siRNA. Furthermore, the LP-0.5F5/2.5P2-CS and LP-0.5F5/2.5P1.6-CL lipoplexes exhibited decreased agglutination with erythrocytes through PEGylation, and markedly decreased the accumulation of siRNA in murine lungs after systemic injection. Finally, systemic injection of LP-0.5F5/2.5P2-CS and LP-0.5F5/2.5P1.6-CL lipoplexes resulted in accumulation of siRNA in KB tumor xenografts. These findings suggest that PEGylation of FA-PEG5000-DSPE-modified siRNA lipoplexes with PEG2000-CS or PEG1600-Chol might improve their systemic stability without the loss of selective transfection activity in tumor cells.

Keywords:  Cationic liposome; PEGylation; folate; folate receptor; gene knock-down; siRNA delivery

DOI:  https://doi.org/10.1080/1061186X.2022.2104860
Drug Deliv. 2022 Dec;29(1): 2375-2385

Transferrin receptor targeting segment T7 containing peptide gene delivery vectors for efficient transfection of brain tumor cells.

Ziyao Kang, Chunlan Zeng, Long Tian, Taoran Wang, Sen Yang, Qin Cheng, Jing Zhang, Qingbin Meng, Changhao Zhang, Zhao Meng.

  Successful gene therapy for brain tumors are often limited by two important factors, the existence of blood brain barrier (BBB) and inefficient transfection of brain tumor cells. In this study, we designed a series of peptide-based gene delivery vectors decorated with T7 segment for binding the transferrin (Tf) receptors which were highly expressed on brain tumor cells, and evaluated their ability of gene delivery. The physicochemical properties of peptide vectors or peptide/DNA complexes were studied as well. The in vitro transfection efficiency was investigated in normal and glioma cell lines. Among these complexes, PT-02/DNA complexes showed the highest transfection efficiency in glioma cells and low cytotoxicity in normal cell lines, and it could transport DNA across the BBB model in vitro. Furthermore, PT-02/DNA could deliver pIRES2-EGFP into the brain site of zebrafish in vivo. The designed peptide vectors offered a promising way for glioma gene therapy.

Keywords:  Peptide vectors; blood brain barrier; gene delivery

DOI:  https://doi.org/10.1080/10717544.2022.2102696
J Biomed Nanotechnol. 2022 Apr 01. 18(4): 986-1000

The Application of Sensitive Nano-Confined Nanoparticle System Using siRNA Targeting Extracellular Signal-Regulated Kinase (ERK)/Mitogen-Activated Protein Kinase (MAPK) Pathway in Gastrointestinal Cancer.

Runda Wu, Shan Tong, Mohammad Saeed Kahrizi, Zheng Zhu, Xin Chen.

In this study we tried to develop a Sensitive Nano-Confined Nanoparticles (S-NCN) system using siRNA against Programmed death-ligand 1 (PDL-1) and Polo Like Kinase 1 (Plk1) to treat gastrointestinal cancer. In this regard, we first synthesized the corresponding materials, prepared the S-NCN system, and verified its functionality. Subsequently, we demonstrated in vitro that S-NCN delivery of siPlk1 could effectively downregulate the expression of Plk1 gene in GC1436 cells and lead to significant apoptosis of tumor cells. Xenografted gastrointestinal cancer model mice were used to evaluate the in vivo efficacy of the nanoparticle. We have demonstrated that the developed S-NCN system can efficiently bind siRNA and deliver it into target tumor cells, solving the main obstacle of siRNA delivery in vivo and effectively silencing target genes with a very potential delivery option.

DOI: https://doi.org/10.1166/jbn.2022.3320
J Nanobiotechnology. 2022 Jul 16. 20(1): 336

pH-sensitive packaging of cationic particles by an anionic block copolymer shell.

Jana I Solomun, Liam Martin, Prosper Mapfumo, Elisabeth Moek, Elias Amro, Friedrich Becker, Stefan Tuempel, Stephanie Hoeppener, K Lenhard Rudolph, Anja Traeger.

  Cationic non-viral vectors show great potential to introduce genetic material into cells, due to their ability to transport large amounts of genetic material and their high synthetic versatility. However, designing materials that are effective without showing toxic effects or undergoing non-specific interactions when applied systemically remains a challenge. The introduction of shielding polymers such as polyethylene glycol (PEG) can enhance biocompatibility and circulation time, however, often impairs transfection efficiency. Herein, a multicomponent polymer system is introduced, based on cationic and hydrophobic particles (P(nBMA46-co-MMA47-co-DMAEMA90), (PBMD)) with high delivery performance and a pH-responsive block copolymer (poly((N-acryloylmorpholine)-b-(2-(carboxy)ethyl acrylamide)) (P(NAM72-b-CEAm74), PNC)) as shielding system, with PNAM as alternative to PEG. The pH-sensitive polymer design promotes biocompatibility and excellent stability at extracellular conditions (pH 7.4) and also allows endosomal escape and thus high transfection efficiency under acidic conditions. PNC shielded particles are below 200 nm in diameter and showed stable pDNA complexation. Further, interaction with human erythrocytes at extracellular conditions (pH 7.4) was prevented, while acidic conditions (pH 6) enabled membrane leakage. The particles demonstrate transfection in adherent (HEK293T) as well as difficult-to-transfect suspension cells (K-562), with comparable or superior efficiency compared to commercial linear poly(ethylenimine) (LPEI). Besides, the toxicity of PNC-shielded particles was significantly minimized, in particular in K-562 cells and erythrocytes. In addition, a pilot in vivo experiment on bone marrow blood cells of mice that were injected with PNC-shielded particles, revealed slightly enhanced cell transfection in comparison to naked pDNA. This study demonstrates the applicability of cationic hydrophobic polymers for transfection of adherent and suspension cells in culture as well as in vivo by co-formulation with pH-responsive shielding polymers, without substantially compromising transfection performance.

Keywords:  Biocompatibility; Charge masking; Core–shell nanoparticles; Layer-by-layer; Stealth polymers

DOI:  https://doi.org/10.1186/s12951-022-01528-0
Toxicol Appl Pharmacol. 2022 Jul 14. pii: S0041-008X(22)00288-5. [Epub ahead of print] 116143

Nonclinical safety evaluation of a novel ionizable lipid for mRNA delivery.

Karine Broudic, Alexander Amberg, Markus Schaefer, Hans-Peter Spirkl, Marie-Clotilde Bernard, Paul Desert.

  mRNA vaccines hold tremendous potential in disease control and prevention for their flexibility with respect to production, application, and design. Recent breakthroughs in mRNA vaccination would have not been possible without major advances in lipid nanoparticles (LNPs) technologies. We developed an LNP containing a novel ionizable cationic lipid, Lipid-1, and three well known excipients. An in silico toxicity hazard assessment for genotoxicity, a genotoxicity assessment, and a dose range finding toxicity study were performed to characterize the safety profile of Lipid-1. The in silico toxicity hazard assessment, utilizing two prediction systems DEREK and Leadscope, did not find any structural alert for mutagenicity and clastogenicity, and prediction in the statistical models were all negative. In addition, applying a read-across approach a structurally very similar compound was tested negative in two in vitro assays confirming the low genotoxicity potential of Lipid-1. A dose range finding toxicity study in rabbits, receiving a single intramuscular injection of either different doses of an mRNA encoding Influenza Hemagglutinin H3 antigen encapsulated in the LNP containing Lipid-1 or the empty LNP, evaluated local tolerance and systemic toxicity during a 2-week observation period. Only rabbits exposed to the vaccine were able to develop a specific IgG response, indicating an appropriate vaccine take. The vaccine was well tolerated up to 250 μg mRNA/injection, which was defined as the No Observed Adverse Effect Level (NOAEL). These results support the use of the LNP containing Lipid-1 as an mRNA delivery system for different vaccine formulations and its deployment into clinical trials.

Keywords:  Dose range finding; Genotoxicity; Lipid nanoparticles; Toxicity hazard assessment; mRNA

DOI:  https://doi.org/10.1016/j.taap.2022.116143
Front Bioeng Biotechnol. 2022 ;10 918368

Hyaluronic Acid-Modified Nanoplatforms as a Vector for Targeted Delivery of Autophagy-Related Gene to the Endometriotic Lesions in Mice.

Mengdan Zhao, Meng Zhang, Qin Yu, Weidong Fei, Tiantian Li, Libo Zhu, Yao Yao, Caihong Zheng, Xinmei Zhang.

  This investigation probed endometriosis treatment using targeted nanoparticles (NPs) to modulate autophagic activity. To that end, a novel form of polymer-based NP gene delivery platform consisting of polyethyleneimine (PEI) conjugated to stearic acid (SA) and nucleotides (DNA/siRNAs) and enclosed by hyaluronic acid (HA) was prepared. CD44 is highly upregulated in cystic lesions, and HA-CD44 binding in this specific nanoplatform was used to achieve targeted drug delivery to CD44-expression endometriotic tissues. The expression of autophagy-related genes was modulated to explore the importance of this process in the development of endometriosis. By inducing autophagic activity, we were able to reduce the size of endometriotic cysts and suppress the development of ectopic endometrium. To further confirm the relationship between autophagic activity and this disease in humans and animals, numbers of autophagic vesicles and autophagic protein expression were assessed in lesion tissue samples from patients, revealing there may be consistency between animal and human data. Overall, these data revealed the ability of this (PEI-SA/DNA) HA gene delivery system to regulate autophagic activity and, thereby, aid in the treatment of endometriosis.

Keywords:  autophagy; endometriosis; gene; nanoparticles; targeted therapy

DOI:  https://doi.org/10.3389/fbioe.2022.918368
ACS Appl Mater Interfaces. 2022 Jul 20.

Redox-Unlockable Nanoparticle-Based MST1 Delivery System to Attenuate Hepatic Steatosis via the AMPK/SREBP-1c Signaling Axis.

Yuhan Li, Jing-Jun Nie, Yuhui Yang, Jianning Li, Jiarui Li, Xianxian Wu, Xing Liu, Da-Fu Chen, Zhiwei Yang, Fu-Jian Xu, Yi Yang.

  To date, few effective treatments have been licensed for nonalcoholic fatty liver disease (NAFLD), which a kind of chronic liver disease. Mammalian sterile 20-like kinase 1 (MST1) is reported to be involved in the development of NAFLD. Thus, we evaluated the suitability of a redox-unlockable polymeric nanoparticle Hep@PGEA vector to deliver MST1 or siMST1 (HCP/MST1 or HCP/siMST1) for NAFLD therapy. The Hep@PGEA vector can efficiently deliver the condensed functional nucleic acids MST1 or siMST1 into NAFLD-affected mouse liver to upregulate or downregulate MST1 expression. The HCP/MST1 complexes significantly improved liver insulin resistance sensitivity and reduced liver damage and lipid accumulation by the AMPK/SREBP-1c pathway without significant adverse events. Instead, HCP/siMST1 delivery exacerbates the NAFLD. The analysis of NAFLD patient samples further clarified the role of MST1 in the development of hepatic steatosis in patients with NAFLD. The MST1-based gene intervention is of considerable potential for clinical NAFLD therapy, and the Hep@PGEA vector provides a promising option for NAFLD gene therapy.

Keywords:  AMPK/SREBP-1c signaling axis; gene therapy; nonalcoholic fatty liver disease (NAFLD); redox-unlockable; self-accelerating release

DOI:  https://doi.org/10.1021/acsami.2c05889
Biomacromolecules. 2022 Jul 21.

Polycation Architecture Affects Complexation and Delivery of Short Antisense Oligonucleotides: Micelleplexes Outperform Polyplexes.

Christian J Grimme, Mckenna G Hanson, Louis G Corcoran, Theresa M Reineke.

Herein, we examine the complexation and biological delivery of a short single-stranded antisense oligonucleotide (ASO) payload with four polymer derivatives that form two architectural variants (polyplexes and micelleplexes): a homopolymer poly(2-dimethylaminoethyl methacrylate) (D), a diblock polymer poly(ethylene glycol)methylether methacrylate-block-poly(2-dimethylaminoethyl methacrylate) (ObD), and two micelle-forming variants, poly(2-dimethylaminoethyl methacrylate)-block-poly(n-butyl methacrylate) (DB) and poly(ethylene glycol)methylether methacrylate-block-poly(2-dimethylaminoethyl methacrylate)-block-poly(n-butyl methacrylate) (ObDB). Both polyplexes and micelleplexes complexed ASOs, and the incorporation of an Ob brush enhances colloidal stability. Micellplexes are templated by the size and shape of the unloaded micelle and that micelle-ASO complexation is not sensitive to formulation/mixing order, allowing ease, versatility, and reproducibility in packaging short oligonucleotides. The DB micelleplexes promoted the largest gene silencing, internalization, and tolerable toxicity while the ObDB micelleplexes displayed enhanced colloidal stability and highly efficient payload trafficking despite having lower cellular uptake. Overall, this work demonstrates that cationic micelles are superior delivery vehicles for ASOs denoting the importance of vehicle architecture in biological performance.

DOI: https://doi.org/10.1021/acs.biomac.2c00338
Angew Chem Int Ed Engl. 2022 Jul 22.

Thiol-Disulfide Exchange as a Route for Endosomal Escape of Polymeric Nanoparticles.

Pintu Kanjilal, Kingshuk Dutta, Sankaran Thayumanavan.

  Endosomal entrapment has remained the major bottleneck for cytosolic delivery of nanoparticle-based delivery systems. Uncovering fundamentally new pathways for endosomal escape is therefore highly sought. Herein, we report that disulfide bonds can enhance endosomal escape through contacts with cellular exofacial thiols, in addition to facilitating cellular uptake. Our results are supported through comparative analysis of polymeric nanogels with variable accessibility to disulfide bonds by placing these functionalities at the core or the shell of the nanogels. The findings here inform future chemical design of delivery vehicles.

Keywords:  Disulfide bond; Endosomal Escape; Endosomal entrapment; Polymeric nanogel

DOI:  https://doi.org/10.1002/anie.202209227
Sci Adv. 2022 Jul 22. 8(29): eabo6406

Polymeric nanoparticles for dual-targeted theranostic gene delivery to hepatocellular carcinoma.

Hannah J Vaughan, Camila G Zamboni, Laboni F Hassan, Nicholas P Radant, Desmond Jacob, Ronnie C Mease, Il Minn, Stephany Y Tzeng, Kathleen L Gabrielson, Pranshu Bhardwaj, Xin Guo, David Francisco, Martin G Pomper, Jordan J Green.

Hepatocellular carcinoma (HCC) develops predominantly in the inflammatory environment of a cirrhotic liver caused by hepatitis, toxin exposure, or chronic liver disease. A targeted therapeutic approach is required to enable cancer killing without causing toxicity and liver failure. Poly(beta-amino-ester) (PBAE) nanoparticles (NPs) were used to deliver a completely CpG-free plasmid harboring mutant herpes simplex virus type 1 sr39 thymidine kinase (sr39) DNA to human HCC cells. Transfection with sr39 enables cancer cell killing with the prodrug ganciclovir and accumulation of 9-(4-18F-fluoro-3-hydroxymethylbutyl)guanine (18F-FHBG) for in vivo imaging. Targeting was achieved using a CpG-free human alpha fetoprotein (AFP) promoter (CpGf-AFP-sr39). Expression was restricted to AFP-producing HCC cells, enabling selective transfection of orthotopic HCC xenografts. CpGf-AFP-sr39 NP treatment resulted in 62% reduced tumor size, and therapeutic gene expression was detectable by positron emission tomography (PET). This systemic nanomedicine achieved tumor-specific delivery, therapy, and imaging, representing a promising platform for targeted treatment of HCC.

DOI: https://doi.org/10.1126/sciadv.abo6406
Acta Pharm Sin B. 2022 Apr;12(4): 2043-2056

The effect of drug loading and multiple administration on the protein corona formation and brain delivery property of PEG-PLA nanoparticles.

Yuyun Tang, Jinchao Gao, Tao Wang, Qian Zhang, Antian Wang, Meng Huang, Renhe Yu, Hongzhuan Chen, Xiaoling Gao.

  The presence of protein corona on the surface of nanoparticles modulates their physiological interactions such as cellular association and targeting property. It has been shown that α-mangostin (αM)-loaded poly(ethylene glycol)-poly(l-lactide) (PEG-PLA) nanoparticles (NP-αM) specifically increased low density lipoprotein receptor (LDLR) expression in microglia and improved clearance of amyloid beta (Aβ) after multiple administration. However, how do the nanoparticles cross the blood‒brain barrier and access microglia remain unknown. Here, we studied the brain delivery property of PEG-PLA nanoparticles under different conditions, finding that the nanoparticles exhibited higher brain transport efficiency and microglia uptake efficiency after αM loading and multiple administration. To reveal the mechanism, we performed proteomic analysis to characterize the composition of protein corona formed under various conditions, finding that both drug loading and multiple dosing affect the composition of protein corona and subsequently influence the cellular uptake of nanoparticles in b.End3 and BV-2 cells. Complement proteins, immunoglobulins, RAB5A and CD36 were found to be enriched in the corona and associated with the process of nanoparticles uptake. Collectively, we bring a mechanistic understanding about the modulator role of protein corona on targeted drug delivery, and provide theoretical basis for engineering brain or microglia-specific targeted delivery system.

Keywords:  Aβ, amyloid beta; BBB, blood‒brain barrier; Brain delivery; CNS, central nervous system; DLS, dynamic light scattering; HCS, high content screening; LDLR, low density lipoprotein receptor; MCI, mild cognitive impairment; Microglia; NP, blank PEG-PLA nanoparticles; NP-corona complexes, nanoparticle-corona complexes; NP-αM, αM-loaded PEG-PLA nanoparticles; Nanoparticles; PEG-PLA; PEG-PLA, poly(ethylene glycol)–poly(l-lactide); Protein corona; TEER, trans-epithelial electrical resistance; TEM, transmission electronic microscope; cou6, coumarin 6; cou6-NPs, cou6-labeled nanoparticles; cou7, coumarin 7; αM, α-mangostin

DOI:  https://doi.org/10.1016/j.apsb.2021.09.029
Sci Adv. 2022 Jul 22. 8(29): eabk3150

Cationic nanoparticles enhance T cell tumor infiltration and antitumor immune responses to a melanoma vaccine.

Rasheid Smith, Emad I Wafa, Sean M Geary, Kareem Ebeid, Suhaila O Alhaj-Suliman, Aliasger K Salem.

In clinical settings, cancer vaccines as monotherapies have displayed limited success compared to other cancer immunotherapeutic treatments. Nanoscale formulations have the ability to increase the efficacy of cancer vaccines by combatting the immunosuppressive nature of the tumor microenvironment. Here, we have synthesized a previously unexplored cationic polymeric nanoparticle formulation using polyamidoamine dendrimers and poly(d,l-lactic-co-glycolic acid) that demonstrate adjuvant properties in vivo. Tumor-challenged mice vaccinated with an adenovirus-based cancer vaccine [encoding tumor-associated antigen (TAA)] and subsequently treated with this nanoparticulate formulation showed significant increases in TAA-specific T cells in the peripheral blood, reduced tumor burden, protection against tumor rechallenge, and a significant increase in median survival. An investigation into cell-based pathways suggests that administration of the nanoformulation at the site of the developing tumor may have created an inflammatory environment that attracted activated TAA-specific CD8+ T cells to the vicinity of the tumor, thus enhancing the efficacy of the vaccine.

DOI: https://doi.org/10.1126/sciadv.abk3150