bims-novged 2023-09-24 papers

bims-novged

Biomed News

on Non-viral vectors for gene delivery

Issue of 2023‒09‒24
twelve papers selected by
the Merkel lab, Ludwig-Maximilians University

TAT decorated siRNA polyplexes for inhalation delivery in anti-asthma therapy.
Inhalation delivery of nucleic acid gene therapies in preclinical drug development.
Differences and Similarities of the Intravenously Administered Lipid Nanoparticles in Three Clinical Trials: Potential Linkage between Lipid Nanoparticles and Extracellular Vesicles.
Intranasal mRNA-LNP vaccination protects hamsters from SARS-CoV-2 infection.
Lipid Nanoparticles Deliver mRNA to the Brain after an Intracerebral Injection.
mRNA lipid nanoparticles induce immune tolerance to treat human diseases.
mRNA nanodelivery systems: targeting strategies and administration routes.
Optimization of HPLCCAD method for simultaneous analysis of different lipids in lipid nanoparticles with analytical QbD.
Nonclinical safety assessment of an mRNA Covid-19 vaccine candidate following repeated administrations and biodistribution.
In vivo self-assembly and delivery of VEGFR2 siRNA-encapsulated small extracellular vesicles for lung metastatic osteosarcoma therapy.
Recent Advances in CRISPR/Cas9 Delivery Approaches for Therapeutic Gene Editing of Stem Cells.
mRNA vaccines in disease prevention and treatment.

Eur J Pharm Sci. 2023 Sep 16. pii: S0928-0987(23)00210-5. [Epub ahead of print]190 106580

TAT decorated siRNA polyplexes for inhalation delivery in anti-asthma therapy.

Salvatore Emanuele Drago, Marta Cabibbo, Emanuela Fabiola Craparo, Gennara Cavallaro.

  In this work, a novel protonable copolymer was designed to deliver siRNA through the inhalation route, as an innovative formulation for the management of asthma. This polycation was synthesized by derivatization of α,β-poly(N-2-hydroxyethyl)D,L-aspartamide (PHEA) first with 1,2-Bis(3-aminopropylamino)ethane (bAPAE) and then with a proper amount of maleimide terminated poly(ethylene glycol) (PEG-MLB), with the aim to increase the superficial hydrophilicity of the system, allowing the diffusion trough the mucus layer. Once the complexation ability of the copolymer has been evaluated, obtaining nanosized polyplexes, polyplexes were functionalized on the surface with a thiolated TAT peptide, a cell-penetrating peptide (CPP), exploiting a thiol-ene reaction. TAT decorated polyplexes result to be highly cytocompatible and able to retain the siRNA with a suitable complexation weight ratio during the diffusion process through the mucus. Despite polyplexes establish weak bonds with the mucin chains, these can diffuse efficiently through the mucin layer and therefore potentially able to reach the bronchial epithelium. Furthermore, through cellular uptake studies, it was possible to observe how the obtained polyplexes penetrate effectively in the cytoplasm of bronchial epithelial cells, where they can reduce IL-8 gene expression, after LPS exposure. In the end, in order to obtain a formulation administrable as an inhalable dry powder, polyplexes were encapsulated in mannitol-based microparticles, by spray freeze drying, obtaining highly porous particles with proper technological characteristics that make them potentially administrable by inhalation route.

Keywords:  Asthma; Polyamine; Polyaspartamide; Polyplexes; TAT; siRNA, STAT6

DOI:  https://doi.org/10.1016/j.ejps.2023.106580
Expert Opin Drug Deliv. 2023 Sep 21.

Inhalation delivery of nucleic acid gene therapies in preclinical drug development.

Conor A Ruzycki, Derek Montoya, Hammad Irshad, Jason Cox, Yue Zhou, Jacob D McDonald, Philip J Kuehl.

  INTRODUCTION: Inhaled gene therapy programs targeting diseases of the lung have seen increasing interest in recent years, though as of yet no product has successfully entered the market. Preclinical research to support such programs is critically important in maximizing the chances of developing successful candidates.AREAS COVERED: Aspects of inhalation delivery of gene therapies are reviewed, with a focus on preclinical research in animal models. Various barriers to inhalation delivery of gene therapies are discussed, including aerosolization stresses, aerosol behavior in the respiratory tract, and disposition processes post-deposition. Important aspects of animal models are considered, including determinations of biologically-relevant determinations of dose and issues related to translatability.
EXPERT OPINION: Development of clinically-efficacious inhaled gene therapies has proven difficult owing to numerous challenges. Fit-for-purpose experimental and analytical methods are necessary for determinations of biologically-relevant doses in preclinical animal models. Further developments in disease-specific animal models may aid in improving translatability of results in future work, and we expect to see accelerated interests in inhalation gene therapies for various diseases. Sponsors, researchers, and regulators are encouraged to engage in early and frequent discussion regarding candidate therapies, and additional dissemination of preclinical methodologies would be of immense value in avoiding common pitfalls.

Keywords:  Alpha-1 antitrypsin deficiency; Asthma; Cystic fibrosis; Dose determination; Inhaled gene therapies; Lung cancer; animal models; preclinical drug development; translatability

DOI:  https://doi.org/10.1080/17425247.2023.2261369
Mol Pharm. 2023 Sep 17.

Differences and Similarities of the Intravenously Administered Lipid Nanoparticles in Three Clinical Trials: Potential Linkage between Lipid Nanoparticles and Extracellular Vesicles.

Yuta Suzuki, Yuri Katsurada, Kenji Hyodo.

  Lipid nanoparticles (LNPs) are clinically validated drug-delivery carriers. However, clinical data on intravenously administered LNPs are limited compared with those on intramuscularly administered LNPs (mRNA vaccines against COVID-19). Here, we reviewed three clinically tested intravenously administered LNPs (patisiran, mRNA-1944, and NTLA-2001). We summarize the differences and similarities in their formulations, mechanisms of action, and pharmacokinetics profiles. In humans, patisiran and mRNA-1944 exhibited similar multiphasic pharmacokinetic profiles with a secondary peak in the RNA concentration. siRNA (patisiran) and mRNA (mRNA-1944) exhibited prolonged blood circulation and were detectable for more than 28 days after a single administration. We further summarize the basics of extracellular vesicles (EVs) and discuss the potential linkages between LNPs and EVs. This Review provides an understanding of the human clinical data of intravenous LNP formulations, which can be potentially explored to develop next-generation LNP-and EV-based drug delivery carriers.

Keywords:  RNA therapeutics; drug delivery; extracellular vesicles; lipid nanoparticles; mRNA

DOI:  https://doi.org/10.1021/acs.molpharmaceut.3c00547
Sci Adv. 2023 Sep 22. 9(38): eadh1655

Intranasal mRNA-LNP vaccination protects hamsters from SARS-CoV-2 infection.

Gabriela Baldeon Vaca, Michelle Meyer, Ana Cadete, Chiaowen Joyce Hsiao, Anne Golding, Albert Jeon, Eric Jacquinet, Emily Azcue, Chenxia Monica Guan, Xavier Sanchez-Felix, Colette A Pietzsch, Chad E Mire, Matthew A Hyde, Margaret E Comeaux, Julie M Williams, Jean C Sung, Andrea Carfi, Darin K Edwards, Alexander Bukreyev, Kapil Bahl.

Intranasal vaccination represents a promising approach for preventing disease caused by respiratory pathogens by eliciting a mucosal immune response in the respiratory tract that may act as an early barrier to infection and transmission. This study investigated immunogenicity and protective efficacy of intranasally administered messenger RNA (mRNA)-lipid nanoparticle (LNP) encapsulated vaccines against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in Syrian golden hamsters. Intranasal mRNA-LNP vaccination systemically induced spike-specific binding [immunoglobulin G (IgG) and IgA] and neutralizing antibodies. Intranasally vaccinated hamsters also had decreased viral loads in the respiratory tract, reduced lung pathology, and prevented weight loss after SARS-CoV-2 challenge. Together, this study demonstrates successful immunogenicity and protection against respiratory viral infection by an intranasally administered mRNA-LNP vaccine.

DOI: https://doi.org/10.1126/sciadv.adh1655
Biochemistry. 2023 Sep 20.

Lipid Nanoparticles Deliver mRNA to the Brain after an Intracerebral Injection.

Jan Tuma, Yu-Ju Chen, Michael G Collins, Abhik Paul, Jie Li, Hesong Han, Rohit Sharma, Niren Murthy, Hye Young Lee.

Neurological disorders are often debilitating conditions with no cure. The majority of current therapies are palliative rather than disease-modifying; therefore, new strategies for treating neurological disorders are greatly needed. mRNA-based therapeutics have great potential for treating such neurological disorders; however, challenges with delivery have limited their clinical potential. Lipid nanoparticles (LNPs) are a promising delivery vector for the brain, given their safer toxicity profile and higher efficacy. Despite this, very little is known about LNP-mediated delivery of mRNA into the brain. Here, we employ MC3-based LNPs and successfully deliver Cre mRNA and Cas9 mRNA/Ai9 sgRNA to the adult Ai9 mouse brain; greater than half of the entire striatum and hippocampus was found to be penetrated along the rostro-caudal axis by direct intracerebral injections of MC3 LNP mRNAs. MC3 LNP Cre mRNA successfully transfected cells in the striatum (∼52% efficiency) and hippocampus (∼49% efficiency). In addition, we demonstrate that MC3 LNP Cas9 mRNA/Ai9 sgRNA edited cells in the striatum (∼7% efficiency) and hippocampus (∼3% efficiency). Further analysis demonstrates that MC3 LNPs mediate mRNA delivery to multiple cell types including neurons, astrocytes, and microglia in the brain. Overall, LNP-based mRNA delivery is effective in brain tissue and shows great promise for treating complex neurological disorders.

DOI: https://doi.org/10.1021/acs.biochem.3c00371
Med Rev (Berl). 2023 Apr;3(2): 180-183

mRNA lipid nanoparticles induce immune tolerance to treat human diseases.

Wei Cao, Tian Xia.

  Rapid developments in the coronavirus disease 2019 (COVID-19) mRNA vaccine showcased the power of lipid nanoparticle (LNP) delivery systems in fighting infectious diseases. In addition, mRNA therapeutics are also in development for cancer immunotherapy. Recently, mRNA therapy has been expanded to induce immune tolerance, the opposite of immune-boosting effects, to treat diseases involving enhanced immune responses including allergies and autoimmune diseases. mRNA LNPs have been used to treat peanut allergy by us and autoimmune experimental autoimmune encephalomyelitis by Ugur Sahin. It is expected that more and more research is going to delve into the immune tolerance field for allergies and autoimmune diseases, where effective therapies are in short supply.

Keywords:  allergy; autoimmune diseases; mRNA lipid nanoparticle; nanomedicine

DOI:  https://doi.org/10.1515/mr-2023-0008
Biomater Res. 2023 Sep 22. 27(1): 90

mRNA nanodelivery systems: targeting strategies and administration routes.

Mujie Yuan, Zeyu Han, Yan Liang, Yong Sun, Bin He, Wantao Chen, Fan Li.

  With the great success of coronavirus disease (COVID-19) messenger ribonucleic acid (mRNA) vaccines, mRNA therapeutics have gained significant momentum for the prevention and treatment of various refractory diseases. To function efficiently in vivo and overcome clinical limitations, mRNA demands safe and stable vectors and a reasonable administration route, bypassing multiple biological barriers and achieving organ-specific targeted delivery of mRNA. Nanoparticle (NP)-based delivery systems representing leading vector approaches ensure the successful intracellular delivery of mRNA to the target organ. In this review, chemical modifications of mRNA and various types of advanced mRNA NPs, including lipid NPs and polymers are summarized. The importance of passive targeting, especially endogenous targeting, and active targeting in mRNA nano-delivery is emphasized, and different cellular endocytic mechanisms are discussed. Most importantly, based on the above content and the physiological structure characteristics of various organs in vivo, the design strategies of mRNA NPs targeting different organs and cells are classified and discussed. Furthermore, the influence of administration routes on targeting design is highlighted. Finally, an outlook on the remaining challenges and future development toward mRNA targeted therapies and precision medicine is provided.

Keywords:  Administration routes; Nanodelivery systems; Targeting; mRNA

DOI:  https://doi.org/10.1186/s40824-023-00425-3
J Chromatogr A. 2023 Sep 16. pii: S0021-9673(23)00600-3. [Epub ahead of print]1709 464375

Optimization of HPLCCAD method for simultaneous analysis of different lipids in lipid nanoparticles with analytical QbD.

Ki Hyun Kim, Ji Eun Lee, Jae Chul Lee, Ravi Maharjan, Hyunsuk Oh, Kyeong Lee, Nam Ah Kim, Seong Hoon Jeong.

  Since lipid nanoparticles (LNP) have emerged as a potent drug delivery system, the objective of this study was to develop and optimize a robust high-performance liquid chromatography with charged aerosol detectors (HPLCCAD) method to simultaneously quantify different lipids in LNPs using the analytical quality by design (AQbD) approach. After defining analytical target profile (ATP), critical method attributes (CMAs) were established as a resolution between the closely eluting lipid peaks and the total analysis time. Thus, potential high-risk method parameters were identified through the initial risk assessment. These parameters were screened using Plackett-Burman design, and three critical method parameters (CMPs)-MeOH ratio, flow rate, and column temperature-were selected for further optimization. Box-Behnken design was employed to develop the quadratic models that explain the relationship between the CMPs and CMAs and to determine the optimal operating conditions. Moreover, to ensure the robustness of the developed method, a method operable design region (MODR) was established using the Monte Carlo simulation. The MODR was identified within the probability map, where the risk of failure to achieve the desired CMAs was less than 1%. The optimized method was validated according to the ICH guidelines (linearity: R2 > 0.995, accuracy: 97.15-100.48% recovery, precision: RSD < 5%) and successfully applied for the analysis of the lipid in the LNP samples. The development of the analytical method to quantify the lipids is essential for the formulation development and quality control of LNP-based drugs since the potency of LNPs is significantly dependent on the compositions and contents of the lipids in the formation.

Keywords:  Analytical quality by design; Charged aerosol detector; Design of experiment; Lipid nanoparticles

DOI:  https://doi.org/10.1016/j.chroma.2023.464375
J Appl Toxicol. 2023 Sep 18.

Nonclinical safety assessment of an mRNA Covid-19 vaccine candidate following repeated administrations and biodistribution.

Karine Broudic, Sébastien Laurent, Vladimir Perkov, Charlene Simon, Marie Garinot, Nathalie Truchot, Julie Latour, Paul Désert.

  Messenger RNA (mRNA) vaccines have demonstrated efficacy against Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) in humans. mRNA technology holds tremendous potential for rapid control and prevention of emergencies due to its flexibility with respect to production, application, and design for an efficacious and safe use in humans. We assessed the toxicity and biodistribution of MRT5500, an mRNA vaccine encoding for the full-length of the SARS-CoV-2 spike protein and delivered by lipid nanoparticles (LNPs) containing a novel ionizable lipid, Lipid-1 in preclinical animal models. In the repeated dose toxicity study, rabbits received three intramuscular (IM) injections of MRT5500 at 3-week interval followed by a 4-week observation period. In an exploratory biodistribution study in mice receiving a single IM injection of an mRNA encoding luciferase encapsulated in an LNP containing Lipid-1, the expression of the luciferase protein was monitored in vivo and ex vivo at several time points. In the regulatory biodistribution study in rabbits receiving a single IM injection of MRT5500, the quantification of the mRNA and the ionizable Lipid-1 were monitored in the same organs and time points as in the exploratory biodistribution study. MRT5500 was safe and well-tolerated with a transient acute phase response/inflammation and an expected vaccine-related immunological response, typical of those observed following a vaccine administration. The biodistribution data demonstrated that the mRNA and Lipid-1 components of the vaccine formulations were mainly detected at the injection site and in the draining lymph nodes. These results support the use of MRT5500 and its deployment into clinical trials.

Keywords:  Covid-19; SARS-CoV-2; biodistribution; lipid nanoparticles; mRNA vaccine; repeated dose toxicity study

DOI:  https://doi.org/10.1002/jat.4548
Cell Death Dis. 2023 Sep 22. 14(9): 626

In vivo self-assembly and delivery of VEGFR2 siRNA-encapsulated small extracellular vesicles for lung metastatic osteosarcoma therapy.

Lingfeng Yu, Gentao Fan, Qingyan Wang, Yan Zhu, Hao Zhu, Jiang Chang, Zhen Wang, Shoubin Zhan, Xianming Hua, Diankun She, Jianhao Huang, Yicun Wang, Jianning Zhao, Chen-Yu Zhang, Xi Chen, Guangxin Zhou.

The prognosis of lung metastatic osteosarcoma (OS) remains disappointing. siRNA-based gene silencing of VEGFR2 is a promising treatment strategy for lung metastatic OS, but there is a lack of safe and efficient delivery systems to encapsulate siRNAs for in vivo administration. This study presented a synthetic biological strategy that remolds the host liver with synthesized genetic circuits for efficient in vivo VEGFR2 siRNA delivery. After being taken-up by hepatocytes, the genetic circuit (in the form of a DNA plasmid) reprogrammed the liver to drive the autonomous intrahepatic assembly and encapsulation of VEGFR2 siRNAs into secretory small extracellular vesicles (sEVs), thus allowing for the transport of self-assembled VEGFR2 siRNAs towards the lung. The results showed that our strategy was superior to the positive medicine (Apatinib) for OS lung metastasis in terms of therapeutic efficacy and toxic adverse effects and may provide a feasible and viable therapeutic solution for lung metastatic OS.

DOI: https://doi.org/10.1038/s41419-023-06159-3
Stem Cell Rev Rep. 2023 Sep 18.

Recent Advances in CRISPR/Cas9 Delivery Approaches for Therapeutic Gene Editing of Stem Cells.

Malihe Lotfi, Dorsa Morshedi Rad, Samaneh Sharif Mashhadi, Atefeh Ashouri, Majid Mojarrad, Sina Mozaffari-Jovin, Shima Farrokhi, Maryam Hashemi, Marzieh Lotfi, Majid Ebrahimi Warkiani, Mohammad Reza Abbaszadegan.

  Rapid advancement in genome editing technologies has provided new promises for treating neoplasia, cardiovascular, neurodegenerative, and monogenic disorders. Recently, the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system has emerged as a powerful gene editing tool offering advantages, including high editing efficiency and low cost over the conventional approaches. Human pluripotent stem cells (hPSCs), with their great proliferation and differentiation potential into different cell types, have been exploited in stem cell-based therapy. The potential of hPSCs and the capabilities of CRISPR/Cas9 genome editing has been paradigm-shifting in medical genetics for over two decades. Since hPSCs are categorized as hard-to-transfect cells, there is a critical demand to develop an appropriate and effective approach for CRISPR/Cas9 delivery into these cells. This review focuses on various strategies for CRISPR/Cas9 delivery in stem cells.

Keywords:  CRISPR/Cas9 system; Delivery technologies; Gene editing; Stem cell therapy; Stem cells

DOI:  https://doi.org/10.1007/s12015-023-10585-3
Signal Transduct Target Ther. 2023 Sep 20. 8(1): 365

mRNA vaccines in disease prevention and treatment.

Gang Zhang, Tianyu Tang, Yinfeng Chen, Xing Huang, Tingbo Liang.

mRNA vaccines have emerged as highly effective strategies in the prophylaxis and treatment of diseases, thanks largely although not totally to their extraordinary performance in recent years against the worldwide plague COVID-19. The huge superiority of mRNA vaccines regarding their efficacy, safety, and large-scale manufacture encourages pharmaceutical industries and biotechnology companies to expand their application to a diverse array of diseases, despite the nonnegligible problems in design, fabrication, and mode of administration. This review delves into the technical underpinnings of mRNA vaccines, covering mRNA design, synthesis, delivery, and adjuvant technologies. Moreover, this review presents a systematic retrospective analysis in a logical and well-organized manner, shedding light on representative mRNA vaccines employed in various diseases. The scope extends across infectious diseases, cancers, immunological diseases, tissue damages, and rare diseases, showcasing the versatility and potential of mRNA vaccines in diverse therapeutic areas. Furthermore, this review engages in a prospective discussion regarding the current challenge and potential direction for the advancement and utilization of mRNA vaccines. Overall, this comprehensive review serves as a valuable resource for researchers, clinicians, and industry professionals, providing a comprehensive understanding of the technical aspects, historical context, and future prospects of mRNA vaccines in the fight against various diseases.

DOI: https://doi.org/10.1038/s41392-023-01579-1