bims-novged 2022-05-15 papers

bims-novged

Biomed News

on Non-viral vectors for gene delivery

Issue of 2022–05–15
ten papers selected by
the Merkel lab, Ludwig-Maximilians University

Optimization of DOTAP/chol Cationic Lipid Nanoparticles for mRNA, pDNA, and Oligonucleotide Delivery.
Estimating the two graph dextran-stearic acid-spermine polymers based on iron oxide nanoparticles as carrier for gene delivery.
Modified chitosan for effective renal delivery of siRNA to treat acute kidney injury.
Enhancing CRISPR/Cas gene editing through modulating cellular mechanical properties for cancer therapy.
Spray drying: Inhalable powders for pulmonary gene therapy.
Virus-like siRNA construct dynamically responsive to sequential microenvironments for potent RNA interference.
Role of polyplex charge density in lipopolyplexes.
pH-responsive co-loaded sulfated hyaluronic acid nanoparticles for treatment of breast cancer via co-targeting tumor cells and CAFs.
Computational study and design of effective siRNAs to silence structural proteins associated genes of Indian SARS-CoV-2 strains.
Poly(l-lactide-co-caprolactone-co-glycolide)-Based Nanoparticles as Delivery Platform: Effect of the Surfactants on Characteristics and Delivery Efficiency.

AAPS PharmSciTech. 2022 May 09. 23(5): 135

Optimization of DOTAP/chol Cationic Lipid Nanoparticles for mRNA, pDNA, and Oligonucleotide Delivery.

Mengwei Sun, Utkarsh J Dang, Yuhao Yuan, Alexandra Maria Psaras, Ositomiwa Osipitan, Tracy A Brooks, Fake Lu, Anthony J Di Pasqua.

  Lipid nanoparticles (LNPs) can be used as delivery vehicles for nucleic acid biotherapeutics. In fact, LNPs are currently being used in the Pfizer/BioNTech and Moderna COVID-19 vaccines. Cationic LNPs composed of 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP)/cholesterol (chol) LNPs have been classified as one of the most efficient gene delivery systems and are being tested in numerous clinical trials. The objective of this study was to examine the effect of the molar ratio of DOTAP/chol, PEGylation, and lipid to mRNA ratio on mRNA transfection, and explore the applications of DOTAP/chol LNPs in pDNA and oligonucleotide transfection. Here we showed that PEGylation significantly decreased mRNA transfection efficiency of DOTAP/chol LNPs. Among non-PEGylated LNP formulations, 1:3 molar ratio of DOTAP/chol in DOTAP/chol LNPs showed the highest mRNA transfection efficiency. Furthermore, the optimal ratio of DOTAP/chol LNPs to mRNA was tested to be 62.5 µM lipid to 1 μg mRNA. More importantly, these mRNA-loaded nanoparticles were stable for 60 days at 4 °C storage without showing reduction in transfection efficacy. We further found that DOTAP/chol LNPs were able to transfect pDNA and oligonucleotides, demonstrating the ability of these LNPs to transport the cargo into the cell nucleus. The influence of various factors in the formulation of DOTAP/chol cationic LNPs is thus described and will help improve drug delivery of nucleic acid-based vaccines and therapies.

Keywords:  DOTAP/cholesterol; LNP-mRNA; lipid nanoparticles; mRNA delivery; transfection efficiency

DOI:  https://doi.org/10.1208/s12249-022-02294-w
Biopolymers. 2022 May 12. e23491

Estimating the two graph dextran-stearic acid-spermine polymers based on iron oxide nanoparticles as carrier for gene delivery.

Mehrnoosh Kazemi-Ashtiyani, Behnam Hajipour-Verdom, Mohammad Satari, Parviz Abdolmaleki, Saman Hosseinkhani, Hossein Shaki.

  Non-viral gene carriers have shown noticeable potential in gene delivery because of limited side effects, biocompatibility, simplicity, and the ability to take advantage of electrostatic interactions. However, the low transfection rate of non-viral vectors under physiological conditions is controversial. This study aimed to decrease the transfection time using a static magnetic field. We used self-assembled cationic polysaccharides based on dextran-stearic acid-spermine (DSASP) conjugates associated with Fe3 O4 superparamagnetic nanoparticles to investigate their potential as gene carriers to promote the target delivery. Our findings illustrate that the magnetic nanoparticles are spherical with a positive surface charge and exhibit superparamagnetic behavior. The DSASP-pDNA/Fe3 O4 complexes offered a strong pDNA condensation, protection against DNase degradation, and significant cell viability in HEK 293T cells. Our results demonstrated that although conjugation of stearic acid could play a role in transfection efficiency, DSASP magnetic carriers with more spermine derivatives showed better affinity between the amphiphilic polymer and the negatively charged cell membrane.

Keywords:  dextran-stearic acid-spermine; gene delivery; iron oxide nanoparticles; non-viral carrier; static magnetic field

DOI:  https://doi.org/10.1002/bip.23491
Biomaterials. 2022 May 02. pii: S0142-9612(22)00202-2. [Epub ahead of print]285 121562

Modified chitosan for effective renal delivery of siRNA to treat acute kidney injury.

Weimin Tang, Sudipta Panja, Chinmay M Jogdeo, Siyuan Tang, Ling Ding, Ao Yu, Kirk W Foster, Del L Dsouza, Yashpal S Chhonker, Heather Jensen-Smith, Hee-Seong Jang, Erika I Boesen, Daryl J Murry, Babu Padanilam, David Oupický.

  Acute kidney injury (AKI) is characterized by a sudden decrease in renal function and impacts growing number of people worldwide. RNA interference (RNAi) showed potential to treat diseases with no or limited conventional therapies, including AKI. Suitable carriers are needed to protect and selectively deliver RNAi to target cells to fully explore this therapeutic modality. Here, we report on the synthesis of chitosan modified with α-cyclam-p-toluic acid (C-CS) as a novel siRNA carrier for targeted delivery to injured kidneys. We demonstrate that conjugation of the α-cyclam-p-toluic acid to chitosan imparts the C-CS polymer with targeting and antagonistic properties to cells overexpressing chemokine receptor CXCR4. In contrast, the parent α-cyclam-p-toluic acid showed no such properties. Self-assembled C-CS/siRNA nanoparticles rapidly accumulate in the injured kidneys and show long retention in renal tubules. Apoptosis and metabolic and inflammatory pathways induced by p53 are important pathological mechanisms in the development of AKI. Nanoparticles with siRNA against p53 (sip53) were formulated and intravenously injected for attenuation of IRI-AKI. Due to the favorable accumulation in injured kidneys, the treatment with C-CS/sip53 decreased renal injury, extent of renal apoptosis, macrophage and neutrophil infiltration, and improved renal function. Overall, our study suggests that C-CS/siRNA nanoparticles have the potential to effectively accumulate and deliver therapeutic siRNAs to injured kidneys through CXCR4 binding, providing a novel way for AKI therapy.

Keywords:  Acute kidney injury; CXCR4; Chitosan; Ischemia-reperfusion injury; Polyplexes; p53

DOI:  https://doi.org/10.1016/j.biomaterials.2022.121562
Nat Nanotechnol. 2022 May 12.

Enhancing CRISPR/Cas gene editing through modulating cellular mechanical properties for cancer therapy.

Di Zhang, Guoxun Wang, Xueliang Yu, Tuo Wei, Lukas Farbiak, Lindsay T Johnson, Alan Mark Taylor, Jiazhu Xu, Yi Hong, Hao Zhu, Daniel J Siegwart.

Genome editing holds great potential for cancer treatment due to the ability to precisely inactivate or repair cancer-related genes. However, delivery of CRISPR/Cas to solid tumours for efficient cancer therapy remains challenging. Here we targeted tumour tissue mechanics via a multiplexed dendrimer lipid nanoparticle (LNP) approach involving co-delivery of focal adhesion kinase (FAK) siRNA, Cas9 mRNA and sgRNA (siFAK + CRISPR-LNPs) to enable tumour delivery and enhance gene-editing efficacy. We show that gene editing was enhanced >10-fold in tumour spheroids due to increased cellular uptake and tumour penetration of nanoparticles mediated by FAK-knockdown. siFAK + CRISPR-PD-L1-LNPs reduced extracellular matrix stiffness and efficiently disrupted PD-L1 expression by CRISPR/Cas gene editing, which significantly inhibited tumour growth and metastasis in four mouse models of cancer. Overall, we provide evidence that modulating the stiffness of tumour tissue can enhance gene editing in tumours, which offers a new strategy for synergistic LNPs and other nanoparticle systems to treat cancer using gene editing.

DOI: https://doi.org/10.1038/s41565-022-01122-3
Mater Sci Eng C Mater Biol Appl. 2021 Dec 09. pii: S0928-4931(21)00741-4. [Epub ahead of print] 112601

Spray drying: Inhalable powders for pulmonary gene therapy.

M Munir, L Jena, V L Kett, N J Dunne, H O McCarthy.

  Gene therapy holds potential in the treatment of many lung pathologies, as indicated by the growing number of clinical trials in recent decades. Pulmonary delivery of gene therapies via inhalation enables localised treatment while the extensive lung surface area promotes enhanced drug absorption. However, loss of nucleic acid integrity during the aerosolisation process, pulmonary clearance, and undesirable drug deposition, pose a major challenge for local delivery. Therefore, the development of nucleic acids into a stable inhalable pharmaceutical preparation would be advantageous. Dry powder inhalers (DPIs) are considered superior compared to nebulisation and pressurised-metered dose inhalers (pMDIs) due to the production of a stable dry formulation, an easy dispensing process, and minimal physical stress. DPIs are commonly produced via spray drying with a range of excipients, solvents, and separation options which can be modified to improve the stability of the nucleic acid cargo. This review details the ideal characteristics for pulmonary delivery and formulation of DPIs for gene therapy to the lungs. The utilisation of spray drying for the production of nucleic acid-containing DPIs is evaluated, with a specific focus on the influence of instrument parameters, the nucleic acid delivery system, and excipients with respect to cargo stability and functionality.

Keywords:  Dry powder inhaler; Gene therapy; Pulmonary delivery; Spray drying

DOI:  https://doi.org/10.1016/j.msec.2021.112601
J Colloid Interface Sci. 2022 May 05. pii: S0021-9797(22)00786-X. [Epub ahead of print]622 938-949

Virus-like siRNA construct dynamically responsive to sequential microenvironments for potent RNA interference.

Yue Wang, Xiujue Zheng, Jun Liu, Li Chen, Qixian Chen, Yan Zhao.

  Cytoplasmic transportation of therapeutic nucleic acids is deemed as an onerous task with aim of precise knockdown towards the targeted genes. Pertaining to the programed functionalities of natural virus in circumventing the biological barriers, we tailored multifaceted chemistries into manufacture of synthetic siRNA delivery vehicles in resembling the functionalities of viral vectors to dynamically tackle with a sequential of biological obstacles encountered in the journey of systemic anti-tumor RNAi therapy. Once harnessing ligands with RGD motif for specific internalization into subcellular endosomal compartments of the tumor cells, the architecture of the proposed delivery vehicles was subjected to facile transformation responsive to pH stimuli in acidic endosomal compartments. The external biocompatible PEGylation palisade was consequently detached, unveiling the cytomembrane-lytic cationic components to commit disruptive potencies to the anionic endosomal membranes for translocation of siRNA conjugates into cytosol. Eventually, liberation of active siRNA could be accomplished due to its responsiveness to the strikingly high level of glutathione in cytosol, thereby contributing to potent RNAi. Hence, our elaborated virus-mimicking platform has demonstrated significant anti-tumor efficacy through systemic administration of anti-angiogenic RNAi payloads, which inspired prosperous potentials in a variety of therapeutic applications.

Keywords:  Anti-angiogenesis; Nanomedicine; Redox responsive; pH responsive; siRNA

DOI:  https://doi.org/10.1016/j.jcis.2022.05.006
Nanoscale. 2022 May 10.

Role of polyplex charge density in lipopolyplexes.

Jianxiang Huang, Wei Song, Lijun Meng, Youqing Shen, Ruhong Zhou.

Lipopolyplexes have received extensive attention lately in gene therapy delivery. However, the interactions between the polyplex and the liposome and their underlying molecular mechanisms remain to be elucidated. Here, we adopted a simple model, mainly to illustrate the impact of polyplex charge density on the self-assembly of liposomes (containing DOPE and CHEMS lipids) using coarse-grained molecular dynamics simulations. Our simulation results show that when the charge density increases in the polyplex, more lipids, especially CHEMS (a negatively charged helper lipid) lipids, are attracted to the polyplex (positively charged) surface, and meanwhile nearby water molecules are driven away from the polyplex, resulting in a less spherical liposome. Energy decomposition analyses further reveal that, at higher charge densities, the polyplex exhibits much stronger interactions with CHEMS lipids than with water molecules, with the majority contribution from electrostatic interactions. In addition, the mobility of lipids, especially CHEMS, is reduced as the polyplex charge density increases, indicating a more rigid liposome. Overall, our molecular dynamics simulations elucidate the influence of polyplex charge density on the liposome self-assembly process at the atomic level, which provides a complementary approach to experiments for a better understanding of this promising gene therapy delivery system.

DOI: https://doi.org/10.1039/d1nr07897f
Recent Pat Anticancer Drug Discov. 2022 May 10.

pH-responsive co-loaded sulfated hyaluronic acid nanoparticles for treatment of breast cancer via co-targeting tumor cells and CAFs.

Di Wang, Jingliang Wu, Cuiping Qi, Jinping Dong, Xueying Ding, Guohua Yu, Shuzhen Liu, Bo Zhang, Zhiqin Gao, Xiuhong Wei, Hongying Liu.

   BACKGROUND: Tumor metastasis is a main cause of death in patients with breast cancer. The cross-talk between cancer associated fibroblasts (CAFs) and tumor cells plays an important role in promoting tumor invasion and metastasis. It is important to develop a novel delivery system to inhibit tumor development by simultaneously targeting both CAFs and tumor cells.
OBJECTIVE: The main objective of this research was to prepare nanoparticles to inhibit tumor proliferation and migration by blocking the cross-talk of tumor-CAFs. Additionally, a novel "MCF-7+NIH/3T3" mixed cell model was established to mimic tumor microenvironment (TME).
METHODS: In this study, the pH-responsive nanoparticles (MIF/DOX-sul-HA NPs) basing on sulfated hyaluronic acid (sul-HA) polymers were prepared for co-delivery of doxorubicin (DOX) and mifepristone (MIF). The effects of anti-proliferation and anti-metastasis of MIF/DOX-sul-HA NPs were investigated both in vitro and in vivo.
RESULTS: The results showed that MIF/DOX-sul-HA NPs were nearly spherical in shape with narrow particle size distribution and pH-responsive drug release, and could be taken up by both of MCF-7 and NIH/3T3 cells. Compared with MCF-7 cells alone, the anti-tumor effect of single DOX was weak in the "MCF-7+NIH/3T3" mixed cell model. MIF/DOX-sul-HA NPs exhibited strong effects of anti-proliferation and anti-metastasis than free single drug.
CONCLUSION: The sul-HA nanoparticles for co-delivery of DOX and MIF could be a promising combined therapy strategy for the treatment of breast cancer.

Keywords:  breast cancer; combination therapy; delivery; nanoparticles

DOI:  https://doi.org/10.2174/1574892817666220510123820
Comput Biol Chem. 2022 Apr 29. pii: S1476-9271(22)00067-6. [Epub ahead of print]98 107687

Computational study and design of effective siRNAs to silence structural proteins associated genes of Indian SARS-CoV-2 strains.

Premnath Madanagopal, Harshini Muthukumar, Kothai Thiruvengadam.

  SARS-CoV-2 is a highly transmissible and pathogenic coronavirus that first emerged in late 2019 and has since triggered a pandemic of acute respiratory disease named COVID-19 which poses a significant threat to all public health institutions in the absence of specific antiviral treatment. Since the outbreak began in March 2020, India has reported 4.77 lakh Coronavirus deaths, according to the World Health Organization (WHO). The innate RNA interference (RNAi) pathway, on the other hand, allows for the development of nucleic acid-based antiviral drugs in which complementary small interfering RNAs (siRNAs) mediate the post-transcriptional gene silencing (PTGS) of target mRNA. Therefore, in this current study, the potential of RNAi was harnessed to construct siRNA molecules that target the consensus regions of specific structural proteins associated genes of SARS-CoV-2, such as the envelope protein gene (E), membrane protein gene (M), nucleocapsid phosphoprotein gene (N), and surface glycoprotein gene (S) which are important for the viral pathogenesis. Conserved sequences of 811 SARS-CoV-2 strains from around India were collected to design 21 nucleotides long siRNA duplex based on various computational algorithms and parameters targeting E, M, N and S genes. The proposed siRNA molecules possessed sufficient nucleotide-based and other features for effective gene silencing and BLAST results revealed that siRNAs' targets have no significant matches across the whole human genome. Hence, siRNAs were found to have no off-target effects on the genome, ruling out the possibility of off-target silencing. Finally, out of 157 computationally identified siRNAs, only 4 effective siRNA molecules were selected for each target gene which is proposed to exert the best action based on GC content, free energy of folding, free energy of binding, melting temperature, heat capacity and molecular docking analysis with Human AGO2 protein. Our engineered siRNA candidates could be used as a genome-level therapeutic treatment against various sequenced SARS-CoV-2 strains in India. However, future applications will necessitate additional validations in vitro and in vivo animal models.

Keywords:  COVID-19; Docking; Gene silencing; RNA interference; SiRNA

DOI:  https://doi.org/10.1016/j.compbiolchem.2022.107687
Nanomaterials (Basel). 2022 May 03. pii: 1550. [Epub ahead of print]12(9):

Poly(l-lactide-co-caprolactone-co-glycolide)-Based Nanoparticles as Delivery Platform: Effect of the Surfactants on Characteristics and Delivery Efficiency.

Magda M Rebanda, Simona Bettini, Laura Blasi, Antonio Gaballo, Andrea Ragusa, Alessandra Quarta, Clara Piccirillo.

  Polymeric nanoparticles made of the copolymer Poly(L-lactide-co-caprolactone-co-glycolide) were prepared using the solvent evaporation method. Two different surfactants, polyvinyl alcohol and dextran, and a mixture of the two were employed. The three types of nanoparticles were used as hosting carriers of two chemotherapeutic drugs, the hydrophilic doxorubicin and the hydrophobic SN-38. The morphostructural characterization showed similar features for the three types of nanoparticles, while the drug encapsulation efficiency indicated that the dextran-based systems are the most effective with both drugs. Cellular studies with breast cancer cells were performed to compare the delivery capability and the cytotoxicity profile of the three nanosystems. The results show that the unloaded nanoparticles are highly biocompatible at the administered concentrations and confirmed that dextran-coated nanoparticles are the most efficient vectors to release the two drugs, exerting cytotoxic activity. PVA, on the other hand, shows limited drug release in vitro, probably due to strong interactions with both drugs. Data also show the release is more efficient for doxorubicin than for SN-38; indeed, the doxorubicin IC50 value for the dextran-coated nanoparticles was about 35% lower than the free drug. This indicates that these nanocarriers are suitable candidates to deliver hydrophilic drugs while needing further modification to host hydrophobic molecules.

Keywords:  SN-38; doxorubicin; polymeric nanoparticle; surfactant-drug interaction

DOI:  https://doi.org/10.3390/nano12091550