bims-drudre 2022-12-18 papers

bims-drudre

Biomed News

on Targeted drug delivery and programmed release mechanisms

Issue of 2022–12–18
24 papers selected by
Ceren Kimna, Technical University of Munich

Delineating the tumour microenvironment response to lipid nanoparticle formulation.
The Extent to Which Lipid Nanoparticles Require Apolipoprotein E and Low-Density Lipoprotein Receptor for Delivery Changes with Ionizable Lipid Structure.
Engineering poly- and micelleplexes for nucleic acid delivery - A reflection on their endosomal escape.
Interaction Kinetics of Individual mRNA-Containing Lipid Nanoparticles with an Endosomal Membrane Mimic: Dependence on pH, Protein Corona Formation, and Lipoprotein Depletion.
Resveratrol-loaded macrophage exosomes alleviate multiple sclerosis through targeting microglia.
Polymeric nano-system for macrophage reprogramming and intracellular MRSA eradication.
Gold Nanobipyramid-Mediated Apoptotic Camouflage of Adipocytes for Obesity Immunotherapy.
Assembly of Fillable Microrobotic Systems by Microfluidic Loading with Dip Sealing.
Multidimensional control of therapeutic human cell function with synthetic gene circuits.
Intracellular Glucose-Depriving Polymer Micelles for Anti-Glycolytic Cancer Treatment.
Dynamic assembly of DNA-ceria nanocomplex in living cells generates artificial peroxisome.
Macrophage Membrane-Coated Nano-Gemcitabine Promotes Lymphocyte Infiltration and Synergizes AntiPD-L1 to Restore the Tumoricidal Function.
Deep Tissue Penetration of Bottle-Brush Polymers via Cell Capture Evasion and Fast Diffusion.
Suppression of flavivirus transmission from animal hosts to mosquitoes with a mosquito-delivered vaccine.
Self-homing nanocarriers for mRNA delivery to the activated hepatic stellate cells in liver fibrosis.
An implantable soft robotic ventilator augments inspiration in a pig model of respiratory insufficiency.
Gene augmentation prevents retinal degeneration in a CRISPR/Cas9-based mouse model of PRPF31 retinitis pigmentosa.
Nanoprobe Based on Biominerals in Protein Corona for Dual-Modality MR Imaging and Therapy of Tumors.
A membrane-free microfluidic approach to mucus permeation for efficient differentiation of mucoadhesive and mucopermeating nanoparticulate systems.
Multimorphic Materials: Spatially Tailoring Mechanical Properties via Selective Initiation of Interpenetrating Polymer Networks.
Protein corona: Friend or foe? Co-opting serum proteins for nanoparticle delivery.
Protein-based nanocages for vaccine development.
Elastin-like polypeptide-based micelles as a promising platform in nanomedicine.
Highly Retina-Permeating and Long-Acting Resveratrol/Metformin Nanotherapeutics for Enhanced Treatment of Macular Degeneration.

J Control Release. 2022 Dec 11. pii: S0168-3659(22)00835-5. [Epub ahead of print]

Delineating the tumour microenvironment response to lipid nanoparticle formulation.

Jessica Ngai, Presley MacMillan, Benjamin R Kingston, Zachary P Lin, Ben Ouyang, Warren C W Chan.

  Nanoparticles can reduce cytotoxicity, increase circulation time and increase accumulation in tumours compared to free drug. However, the value of using nanoparticles for carrying small molecules to treat tumours at the cellular level has been poorly established. Here we conducted a cytodistribution analysis on Doxorubicin-treated and Doxil-treated tumours to delineate the differences between the small molecule therapeutic Doxorubicin and its packaged liposomal Doxil. We found that Doxil kills more cancer cells, macrophages and neutrophils in the 4 T1 breast cancer tumour model, but there is an apparent delayed killing compared to its small molecule counterpart Doxorubicin. This was because the cellular interaction with Doxil has slower uptake kinetics and the particles must be degraded to release the drug and kill the cells. We also found that macrophages and neutrophils in Doxil-treated tumours repopulated the tumour faster than cancer cells during the relapse phase. While researchers conventionally use tumour volume and animal survival to determine a therapeutic effect, our results show diverse cell killing and a greater amount of cell death in vivo after Doxil liposomes are administered. We conclude that the fate and behaviour of the nanocarrier influence its effectiveness as a cancer therapy. Further investigations on the interactions between different nanoparticle designs and the tumour microenvironment components will lead to more precise engineering of nanocarriers to selectively kill tumour cells and prolong the therapeutic effect.

Keywords:  Cancer; Cytodistribution; Doxil; Doxorubicin; Flow cytometry; Liposomes

DOI:  https://doi.org/10.1016/j.jconrel.2022.12.021
Nano Lett. 2022 Dec 15.

The Extent to Which Lipid Nanoparticles Require Apolipoprotein E and Low-Density Lipoprotein Receptor for Delivery Changes with Ionizable Lipid Structure.

Kalina Paunovska, Alejandro J Da Silva Sanchez, Melissa P Lokugamage, David Loughrey, Elisa Schrader Echeverri, Ana Cristian, Marine Z C Hatit, Philip J Santangelo, Kun Zhao, James E Dahlman.

  Lipid nanoparticles (LNPs) have delivered therapeutic RNA to hepatocytes in humans. Adsorption of apolipoprotein E (ApoE) onto these clinical LNP-mRNA drugs has been shown to facilitate hepatocyte entry via the low-density lipoprotein receptor (LDLR). Since ApoE-LDLR trafficking is conserved in mice, non-human primates, and humans, characterizing this mechanism eased clinical transition. Recently, LNPs have delivered mRNA to non-hepatocytes in mice and non-human primates, suggesting they can target new cell types via ApoE- and LDLR-independent pathways. To test this hypothesis, we quantified how 60 LNPs delivered mRNA with cell type resolution in wild-type mice and three knockout mouse strains related to lipid trafficking: ApoE-/-, LDLR-/-, and PCSK9-/-. These data suggest that the hydrophobic tail length of diketopiperazine-based lipids can be changed to drive ApoE- and LDLR-independent delivery in vivo. More broadly, the results support the hypothesis that endogenous LNP trafficking can be tuned by modifying lipid chemistry.

Keywords:  DNA barcoding; LNP; ionizable lipids; knockout; lipid nanoparticle; mRNA

DOI:  https://doi.org/10.1021/acs.nanolett.2c03741
J Control Release. 2022 Dec 09. pii: S0168-3659(22)00822-7. [Epub ahead of print]353 518-534

Engineering poly- and micelleplexes for nucleic acid delivery - A reflection on their endosomal escape.

Benjamin Winkeljann, David C Keul, Olivia M Merkel.

  For the longest time, the field of nucleic acid delivery has remained skeptical whether or not polycationic drug carrier systems would ever make it into clinical practice. Yet, with the disclosure of patents on polyethyleneimine-based RNA carriers through leading companies in the field of nucleic acid therapeutics such as BioNTech SE and the progress in clinical studies beyond phase I trials, this aloofness seems to regress. As one of the most striking characteristics of polymer-based vectors, the extraordinary tunability can be both a blessing and a curse. Yet, knowing about the adjustment screws and how they impact the performance of the drug carrier provides the formulation scientist committed to its development with a head start. Here, we equip the reader with a toolbox - a toolbox that should advise and support the developer to conceptualize a cutting-edge poly- or micelleplex system for the delivery of therapeutic nucleic acids; to be specific, to engineer the vector towards maximum endosomal escape performance at minimum toxicity. Therefore, after briefly sketching the boundary conditions of polymeric vector design, we will dive into the topic of endosomal trafficking. We will not only discuss the most recent knowledge of the endo-lysosomal compartment but further depict different hypotheses and mechanisms that facilitate the endosomal escape of polyplex systems. Finally, we will combine the different facets introduced in the previous chapters with the fundamental building blocks of polymer vector design and evaluate the advantages and drawbacks. Throughout the article, a particular focus will be placed on cellular peculiarities, not only as an additional barrier, but also to give inspiration to how such cell-specific traits might be capitalized on.

Keywords:  Nonviral vector; Polyethyleneimine; Proton sponge hypothesis; RNA; Vaccine

DOI:  https://doi.org/10.1016/j.jconrel.2022.12.008
ACS Nano. 2022 Dec 13.

Interaction Kinetics of Individual mRNA-Containing Lipid Nanoparticles with an Endosomal Membrane Mimic: Dependence on pH, Protein Corona Formation, and Lipoprotein Depletion.

Nima Aliakbarinodehi, Audrey Gallud, Mokhtar Mapar, Emelie Wesén, Sahar Heydari, Yujia Jing, Gustav Emilsson, Kai Liu, Alan Sabirsh, Vladimir P Zhdanov, Lennart Lindfors, Elin K Esbjörner, Fredrik Höök.

  Lipid nanoparticles (LNPs) have emerged as potent carriers for mRNA delivery, but several challenges remain before this approach can offer broad clinical translation of mRNA therapeutics. To improve their efficacy, a better understanding is required regarding how LNPs are trapped and processed at the anionic endosomal membrane prior to mRNA release. We used surface-sensitive fluorescence microscopy with single LNP resolution to investigate the pH dependency of the binding kinetics of ionizable lipid-containing LNPs to a supported endosomal model membrane. A sharp increase of LNP binding was observed when the pH was lowered from 6 to 5, accompanied by stepwise large-scale LNP disintegration. For LNPs preincubated in serum, protein corona formation shifted the onset of LNP binding and subsequent disintegration to lower pH, an effect that was less pronounced for lipoprotein-depleted serum. The LNP binding to the endosomal membrane mimic was observed to eventually become severely limited by suppression of the driving force for the formation of multivalent bonds during LNP attachment or, more specifically, by charge neutralization of anionic lipids in the model membrane due to their association with cationic lipids from earlier attached LNPs upon their disintegration. Cell uptake experiments demonstrated marginal differences in LNP uptake in untreated and lipoprotein-depleted serum, whereas lipoprotein-depleted serum increased mRNA-controlled protein (eGFP) production substantially. This complies with model membrane data and suggests that protein corona formation on the surface of the LNPs influences the nature of the interaction between LNPs and endosomal membranes.

Keywords:  endosomal membrane; ionizable lipid nanoparticle; lipoprotein; mRNA delivery; protein corona

DOI:  https://doi.org/10.1021/acsnano.2c04829
J Control Release. 2022 Dec 14. pii: S0168-3659(22)00840-9. [Epub ahead of print]353 675-684

Resveratrol-loaded macrophage exosomes alleviate multiple sclerosis through targeting microglia.

Xue Zheng, Ke Sun, Yonghui Liu, Xiaona Yin, Haomiao Zhu, Fan Yu, Wei Zhao.

  Despite exosome promise as endogenous drug delivery vehicles, the current understanding of exosome may be insufficient to develop their various applications. Here we synthesized five sialic acid analogues with different length N-acyl side chains and screened out the optimal metabolic precursor for exosome labeling via bio-orthogonal click chemistry. In proof-of-principle labeling experiments, exosomes derived from macrophages (RAW-Exo) strongly co-localized with central nervous system (CNS) microglia. Inspired by this discovery, we developed a resveratrol-loaded RAW-Exo formulation (RSV&Exo) for multiple sclerosis (MS) treatment. Intranasal administration of RSV&Exo significantly inhibited inflammatory responses in the CNS and peripheral system in a mouse model of MS and effectively improved the clinical evolution of MS in vivo. These findings suggested the feasibility and efficacy of engineered RSV&Exo administration for MS, providing a potential therapeutic strategy for CNS diseases.

Keywords:  Click chemistry; Exosomes; Multiple sclerosis; Resveratrol; Sialic acid

DOI:  https://doi.org/10.1016/j.jconrel.2022.12.026
J Control Release. 2022 Dec 12. pii: S0168-3659(22)00828-8. [Epub ahead of print]353 591-610

Polymeric nano-system for macrophage reprogramming and intracellular MRSA eradication.

Yun-Jian Yu, Jian-Hua Yan, Qi-Wen Chen, Ji-Yan Qiao, Si-Yuan Peng, Han Cheng, Meiwan Chen, Xian-Zheng Zhang.

  Intracellular Methicillin-Resistant Staphylococcus aureus (MRSA) remains a major factor of refractory and recurrent infections, which cannot be well addressed by antibiotic therapy. Here, we design a cellular infectious microenvironment-activatable polymeric nano-system to mediate targeted intracellular drug delivery for macrophage reprogramming and intracellular MRSA eradication. The polymeric nano-system is composed of a ferrocene-decorated polymeric nanovesicle formulated from poly(ferrocenemethyl methacrylate)-block-poly(2-methacryloyloxyethyl phosphorylcholine) (PFMMA-b-PMPC) copolymer with co-encapsulation of clofazimine (CFZ) and interferon-γ (IFN-γ). The cellular-targeting PMPC motifs render specific internalization by macrophages and allow efficient intracellular accumulation. Following the internalization, the ferrocene-derived polymer backbone sequentially undergoes hydrophobic-to-hydrophilic transition, charge reversal and Fe release in response to intracellular hydrogen peroxide over-produced upon infection, eventually triggering endosomal escape and on-site cytosolic drug delivery. The released IFN-γ reverses the immunosuppressive status of infected macrophages by reprogramming anti-inflammatory M2 to pro-inflammatory M1 phenotype. Meanwhile, intracellular Fe2+-mediated Fenton reaction together with antibiotic CFZ contributes to increased intracellular hydroxyl radical (•OH) generation. Ultimately, the nano-system achieves robust potency in ablating intracellular MRSA and antibiotic-tolerant persisters by synchronous immune modulation and efficient •OH killing, providing an innovative train of thought for intracellular MRSA control.

Keywords:  Drug delivery; Intracellular MRSA; Macrophage reprogramming; Persister; Polymeric nanovesicle

DOI:  https://doi.org/10.1016/j.jconrel.2022.12.014
Adv Mater. 2022 Dec 11. e2207686

Gold Nanobipyramid-Mediated Apoptotic Camouflage of Adipocytes for Obesity Immunotherapy.

Jiao Yan, Yanjing Wang, Zhengzhi Mu, Xiaoqing Han, Luopeng Bi, Xingbo Wang, Panpan Song, Yaqing Kang, Lulu Wang, Xueyan Zhang, Yanbo Wang, Haiyuan Zhang.

  Obesity treatment is a global public health challenge due to the inadequate weight loss and weight regain even after endeavors with multimodal treatments. Considering abundant resident macrophages in adipose tissues, precise regulation of the interactions between macrophages and adipocytes may provide chances for immunotherapy of obesity. Herein, inspired by the phagocytosis of macrophages to clear apoptotic cells in homeostasis, an immunotherapy strategy for obesity treatment was proposed for the first time through apoptotic camouflage of adipocytes by PA Au BPs to activate macrophages for clearance, where PA Au BPs were gold nanobipyramids engineered with adipose-targeting and apoptotic cell-mimicking functions. During the clearance, macrophage was switched from pro-inflammatory M1 to anti-inflammatory M2, remarkably modulating the immune microenvironment of adipose tissues to prevent weight regain. After inguinal injection with PA Au BPs, the body weights of obesity mice were effectively decreased by 24.4%, and could be decreased by 33.3% when combined with photothermal lipolysis, and little weight regain was associated with treatments. This study demonstrates the strategy of camouflaging adipocytes with apoptotic feature holds great potential for obesity immunotherapy. This article is protected by copyright. All rights reserved.

Keywords:  Apoptotic camouflage; Immunotherapy; Nanoparticles; Obesity; Surface engineering

DOI:  https://doi.org/10.1002/adma.202207686
Adv Mater. 2022 Dec 11. e2207791

Assembly of Fillable Microrobotic Systems by Microfluidic Loading with Dip Sealing.

Rujie Sun, Xin Song, Kun Zhou, Yuyang Zuo, Richard Wang, Omar Rifaie-Graham, David Peeler, Ruoxiao Xie, Yixuan Leng, Hongya Geng, Giulia Brachi, Yun Ma, Yutong Liu, Lorna Barron, Molly M Stevens.

  Microrobots can provide spatiotemporally well-controlled cargo delivery that can improve therapeutic efficiency compared to conventional drug delivery strategies. Robust microfabrication methods to expand the variety of materials or cargoes that can be incorporated into microrobots could greatly broaden the scope of their functions. However, current surface coating or direct blending techniques used for cargo loading result in inefficient loading and poor cargo protection during transportation, which leads to cargo waste, degradation, and non-specific release. Herein, we present a versatile platform to fabricate fillable microrobots using Microfluidic Loading and Dip Sealing (MLDS). MLDS enables the encapsulation of different types of cargoes within hollow microrobots and protection of cargo integrity. The technique is supported by high-resolution 3D printing with an integrated microfluidic loading system, which realises a highly precise loading process and improves cargo loading capacity. A corresponding dip sealing strategy was developed to encase and protect the loaded cargo whilst maintaining the geometric and structural integrity of the loaded microrobots. This dip sealing technique is suitable for different materials, including thermal and light responsive materials. The MLDS platform provides new opportunities for microrobotic systems in targeted drug delivery, environmental sensing, and chemically powered micromotor applications. This article is protected by copyright. All rights reserved.

Keywords:  environmental sensing; fillable microrobotics; microfluidics; micromotor; targeted delivery

DOI:  https://doi.org/10.1002/adma.202207791
Science. 2022 Dec 16. 378(6625): 1227-1234

Multidimensional control of therapeutic human cell function with synthetic gene circuits.

Hui-Shan Li, Divya V Israni, Keith A Gagnon, Kok Ann Gan, Michael H Raymond, Jeffry D Sander, Kole T Roybal, J Keith Joung, Wilson W Wong, Ahmad S Khalil.

Synthetic gene circuits that precisely control human cell function could expand the capabilities of gene- and cell-based therapies. However, platforms for developing circuits in primary human cells that drive robust functional changes in vivo and have compositions suitable for clinical use are lacking. Here, we developed synthetic zinc finger transcription regulators (synZiFTRs), which are compact and based largely on human-derived proteins. As a proof of principle, we engineered gene switches and circuits that allow precise, user-defined control over therapeutically relevant genes in primary T cells using orthogonal, US Food and Drug Administration-approved small-molecule inducers. Our circuits can instruct T cells to sequentially activate multiple cellular programs such as proliferation and antitumor activity to drive synergistic therapeutic responses. This platform should accelerate the development and clinical translation of synthetic gene circuits in diverse human cell types and contexts.

DOI: https://doi.org/10.1126/science.ade0156
Adv Mater. 2022 Dec 16. e2207342

Intracellular Glucose-Depriving Polymer Micelles for Anti-Glycolytic Cancer Treatment.

Jangwook Lee, Kwangmeyung Kim, Ick Chan Kwon, Kuen Yong Lee.

  A new anti-cancer strategy to exploit abnormal metabolism of cancer cells rather than to merely control the drug release or rearrange the tumor microenvironment is reported herein. An anti-glycolytic amphiphilic polymer, designed considering the unique metabolism of cancer cells (Warburg effect) and aimed at the regulation of glucose metabolism, is synthesized through chemical conjugation between glycol chitosan (GC) and phenylboronic acid (PBA). GC-PBA derivatives form stable micellar structures under physiological conditions and respond to changes in glucose concentration. Once the micelles accumulate at the tumor site, intracellular glucose capture occurs, and the resultant energy deprivation through the inhibition of aerobic glycolysis remarkably suppresses tumor growth without significant side effects in vivo. This strategy highlights the need to develop safe and effective cancer treatment without the use of conventional anti-cancer drugs. This article is protected by copyright. All rights reserved.

Keywords:  apoptosis; cancer therapy; glucose-responsive polymer; induced energy deficiency; nutrient deprivation

DOI:  https://doi.org/10.1002/adma.202207342
Nat Commun. 2022 Dec 14. 13(1): 7739

Dynamic assembly of DNA-ceria nanocomplex in living cells generates artificial peroxisome.

Chi Yao, Yuwei Xu, Jianpu Tang, Pin Hu, Hedong Qi, Dayong Yang.

Intracellular accumulation of reactive oxygen species (ROS) leads to oxidative stress, which is closely associated with many diseases. Introducing artificial organelles to ROS-imbalanced cells is a promising solution, but this route requires nanoscale particles for efficient cell uptake and micro-scale particles for long-term cell retention, which meets a dilemma. Herein, we report a deoxyribonucleic acid (DNA)-ceria nanocomplex-based dynamic assembly system to realize the intracellular in-situ construction of artificial peroxisomes (AP). The DNA-ceria nanocomplex is synthesized from branched DNA with i-motif structure that responds to the acidic lysosomal environment, triggering transformation from the nanoscale into bulk-scale AP. The initial nanoscale of the nanocomplex facilitates cellular uptake, and the bulk-scale of AP supports cellular retention. AP exhibits enzyme-like catalysis activities, serving as ROS eliminator, scavenging ROS by decomposing H2O2 into O2 and H2O. In living cells, AP efficiently regulates intracellular ROS level and resists GSH consumption, preventing cells from redox dyshomeostasis. With the protection of AP, cytoskeleton integrity, mitochondrial membrane potential, calcium concentration and ATPase activity are maintained under oxidative stress, and thus the energy of cell migration is preserved. As a result, AP inhibits cell apoptosis, reducing cell mortality through ROS elimination.

DOI: https://doi.org/10.1038/s41467-022-35472-2
ACS Nano. 2022 Dec 16.

Macrophage Membrane-Coated Nano-Gemcitabine Promotes Lymphocyte Infiltration and Synergizes AntiPD-L1 to Restore the Tumoricidal Function.

Jie Li, Yao Wu, Jiaoying Wang, Xiaoxuan Xu, Ao Zhang, Yaping Li, Zhiwen Zhang.

  The limited lymphocyte infiltration and exhaustion of tumoricidal functions in solid tumors remain a formidable obstacle to cancer immunotherapy. Herein, we designed a macrophage membrane-coated nano-gemcitabine system (MNGs) to promote lymphocyte infiltration and then synergized anti-programmed death ligand 1 (antiPD-L1) to reinvigorate the exhausted lymphocytes. MNGs exhibited effective intratumor-permeating and responsive drug-releasing capacity, produced notable elimination of versatile immunosuppressive cells, and promoted lymphocyte infiltration into cancer cell regions in tumors, but over 50% of these infiltrated lymphocytes were in the exhausted state. Compared with MNG monotherapy, the MNGs+antiPD-L1 combination produced 31.77% and 30.63% reduction of exhausted CD3+CD8+ T cells and natural killer (NK) cells and 2.83- and 3.17-fold increases of interferon-γ (IFN-γ)-positive subtypes, respectively, thereby resulting in considerable therapeutic benefits in several tumor models. Thus, MNGs provide an encouraging strategy to promote lymphocyte infiltration and synergize antiPD-L1 to restore their tumoricidal function for cancer immunotherapy.

Keywords:  gemcitabine; immunotherapy; lymphocytes infiltration; macrophage; nanoparticle

DOI:  https://doi.org/10.1021/acsnano.2c07861
ACS Nano. 2022 Dec 14.

Deep Tissue Penetration of Bottle-Brush Polymers via Cell Capture Evasion and Fast Diffusion.

Jean-Michel Rabanel, Marziye Mirbagheri, Mateusz Olszewski, Guojun Xie, Marine Le Goas, Pierre-Luc Latreille, Hermine Counil, Vincent Hervé, Rummenigge Oliveira Silva, Charlotte Zaouter, Vahid Adibnia, Mariana Acevedo, Marc J Servant, Vincent A Martinez, Shunmoogum A Patten, Krzysztof Matyjaszewski, Charles Ramassamy, Xavier Banquy.

  Drug nanocarriers (NCs) capable of crossing the vascular endothelium and deeply penetrating into dense tissues of the CNS could potentially transform the management of neurological diseases. In the present study, we investigated the interaction of bottle-brush (BB) polymers with different biological barriers in vitro and in vivo and compared it to nanospheres of similar composition. In vitro internalization and permeability assays revealed that BB polymers are not internalized by brain-associated cell lines and translocate much faster across a blood-brain barrier model compared to nanospheres of similar hydrodynamic diameter. These observations performed under static, no-flow conditions were complemented by dynamic assays performed in microvessel arrays on chip and confirmed that BB polymers can escape the vasculature compartment via a paracellular route. BB polymers injected in mice and zebrafish larvae exhibit higher penetration in brain tissues and faster extravasation of microvessels located in the brain compared to nanospheres of similar sizes. The superior diffusivity of BBs in extracellular matrix-like gels combined with their ability to efficiently cross endothelial barriers via a paracellular route position them as promising drug carriers to translocate across the blood-brain barrier and penetrate dense tissue such as the brain, two unmet challenges and ultimate frontiers in nanomedicine.

Keywords:  blood-brain barrier; bottle-brush polymer; mouse; vessel-on-a-chip; zebrafish

DOI:  https://doi.org/10.1021/acsnano.2c10554
Nat Commun. 2022 Dec 16. 13(1): 7780

Suppression of flavivirus transmission from animal hosts to mosquitoes with a mosquito-delivered vaccine.

Dan Wen, Limin S Ding, Yanan Zhang, Xiaoye Li, Xing Zhang, Fei Yuan, Tongbiao Zhao, Aihua Zheng.

Zoonotic viruses circulate in the natural reservoir and sporadically spill over into human populations, resulting in endemics or pandemics. We previously found that the Chaoyang virus (CYV), an insect-specific flavivirus (ISF), is replication-defective in vertebrate cells. Here, we develope a proof-of-concept mosquito-delivered vaccine to control the Zika virus (ZIKV) within inaccessible wildlife hosts using CYV as the vector. The vaccine is constructed by replacing the pre-membrane and envelope (prME) proteins of CYV with those of ZIKV, assigned as CYV-ZIKV. CYV-ZIKV replicates efficiently in Aedes mosquitoes and disseminates to the saliva, with no venereal or transovarial transmission observed. To reduce the risk of CYV-ZIKV leaking into the environment, mosquitoes are X-ray irradiated to ensure 100% infertility, which does not affect the titer of CYV-ZIKV in the saliva. Immunization of mice via CYV-ZIKV-carrying mosquito bites elicites robust and persistent ZIKV-specific immune responses and confers complete protection against ZIKV challenge. Correspondingly, the immunized mice could no longer transmit the challenged ZIKV to naïve mosquitoes. Therefore, immunization with an ISF-vectored vaccine via mosquito bites is feasible to induce herd immunity in wildlife hosts of ZIKV. Our study provides a future avenue for developing a mosquito-delivered vaccine to eliminate zoonotic viruses in the sylvatic cycle.

DOI: https://doi.org/10.1038/s41467-022-35407-x
J Control Release. 2022 Dec 12. pii: S0168-3659(22)00834-3. [Epub ahead of print]

Self-homing nanocarriers for mRNA delivery to the activated hepatic stellate cells in liver fibrosis.

Mahmoud A Younis, Yusuke Sato, Yaser H A Elewa, Yasuhiro Kon, Hideyoshi Harashima.

  Herein, we report on the development of a platform for the selective delivery of mRNA to the hard-to-transfect Activated Hepatic Stellate Cells (aHSCs), the fundamental player in the progression of liver fibrosis. Using a microfluidic device (iLiNP), we prepared a series of lipid nanoparticles (LNPs) based on a diverse library of pH-sensitive lipids. After an in-depth in vivo optimization of the LNPs, their mRNA delivery efficiency, selectivity, potency, robustness, and biosafety were confirmed. Furthermore, some mechanistic aspects of their selective delivery to aHSCs were investigated. We identified a promising lipid candidate, CL15A6, that has a high affinity to aHSCs. Tweaking the composition and physico-chemical properties of the LNPs enabled the robust and ligand-free mRNA delivery to aHSCs in vivo post intravenous administration, with a high biosafety at mRNA doses of up to 2 mg/Kg, upon either acute or chronic administrations. The mechanistic investigation suggested that CL15A6 LNPs were taken up by aHSCs via Clathrin-mediated endocytosis through the Platelet-derived growth factor receptor beta (PDGFRβ) and showed a pKa-dependent cellular uptake. The novel and scalable platform reported in this study is highly promising for clinical applications.

Keywords:  Hepatic stellate cells; Lipid nanoparticles; Liver fibrosis; Microfluidic device; mRNA

DOI:  https://doi.org/10.1016/j.jconrel.2022.12.020
Nat Biomed Eng. 2022 Dec 12.

An implantable soft robotic ventilator augments inspiration in a pig model of respiratory insufficiency.

Lucy Hu, Jean Bonnemain, Mossab Y Saeed, Manisha Singh, Diego Quevedo Moreno, Nikolay V Vasilyev, Ellen T Roche.

Severe diaphragm dysfunction can lead to respiratory failure and to the need for permanent mechanical ventilation. Yet permanent tethering to a mechanical ventilator through the mouth or via tracheostomy can hinder a patient's speech, swallowing ability and mobility. Here we show, in a porcine model of varied respiratory insufficiency, that a contractile soft robotic actuator implanted above the diaphragm augments its motion during inspiration. Synchronized actuation of the diaphragm-assist implant with the native respiratory effort increased tidal volumes and maintained ventilation flow rates within the normal range. Robotic implants that intervene at the diaphragm rather than at the upper airway and that augment physiological metrics of ventilation may restore respiratory performance without sacrificing quality of life.

DOI: https://doi.org/10.1038/s41551-022-00971-6
Nat Commun. 2022 Dec 13. 13(1): 7695

Gene augmentation prevents retinal degeneration in a CRISPR/Cas9-based mouse model of PRPF31 retinitis pigmentosa.

Zhouhuan Xi, Abhishek Vats, José-Alain Sahel, Yuanyuan Chen, Leah C Byrne.

Mutations in PRPF31 cause autosomal dominant retinitis pigmentosa, an untreatable form of blindness. Gene therapy is a promising treatment for PRPF31-retinitis pigmentosa, however, there are currently no suitable animal models in which to develop AAV-mediated gene augmentation. Here we establish Prpf31 mutant mouse models using AAV-mediated CRISPR/Cas9 knockout, and characterize the resulting retinal degeneration phenotype. Mouse models with early-onset morphological and functional impairments like those in patients were established, providing new platforms in which to investigate pathogenetic mechanisms and develop therapeutic methods. AAV-mediated PRPF31 gene augmentation restored the retinal structure and function in a rapidly degenerating mouse model, demonstrating the first in vivo proof-of-concept for AAV-mediated gene therapy to treat PRPF31-retinitis pigmentosa. AAV-CRISPR/Cas9-PRPF31 knockout constructs also mediated efficient PRPF31 knockout in human and non-human primate retinal explants, laying a foundation for establishing non-human primate models using the method developed here.

DOI: https://doi.org/10.1038/s41467-022-35361-8
ACS Nano. 2022 Dec 16.

Nanoprobe Based on Biominerals in Protein Corona for Dual-Modality MR Imaging and Therapy of Tumors.

Peisen Zhang, Yuanyuan Qiao, Lichong Zhu, Meng Qin, Qilong Li, Chuang Liu, Yuping Xu, Xin Zhang, Zhihua Gan, Yi Hou.

  Various functional nanomaterials have been fabricated as diagnostic and therapeutic nanomedicines; however, the nanoparticles closely interact with proteins when immersed in biological fluids, forming a "protein corona" that critically alters the biological identity of nanomedicine. Here, we developed a robust strategy to construct theranostic nanoprobes based on protein-corona-coated Fe3O4 nanoparticles and biomineralization in the corona. Water-soluble carboxylic Fe3O4 nanoparticles were prepared by treating oleate-capped Fe3O4 nanoparticles with Lemieux-von Rudloff reagent. Bovine serum albumin (BSA) was used as a model protein to form a corona on the surface of Fe3O4 nanoparticles, endowing the Fe3O4 nanoparticles with biocompatibility and nonimmunogenicity. The protein corona also provides a template for biomimetic mineralization of Fe3+ with tannic acid (TA) to construct Fe3O4@BSA-TAFeIII nanoprobes. The TA-Fe(III) biominerals can not only act as photothermal therapy agents but also interact with unsaturated transferrin in plasma to form a "hybrid" corona, enabling the nanoprobes to target tumor cells through the mediation of transferrin receptors, which commonly overexpress on tumor cell membranes. Once taken in by tumor cells, the protonation of phenol hydroxyl groups in acidic lysosomes would lead to the release of Fe3+, inducing tumor cell death through a ferroptosis/apoptosis hybrid pathway. In addition, the released Fe3+ can boost the T1-weighted MR imaging performance, and the Fe3O4 nanoparticles serve as T2-weighted MR imaging contrast agents. It is thus believed that the current nanoprobes can realize the enhanced dual-modality MR imaging and combined therapy of tumors through controlling the protein corona and biomineralization.

Keywords:  biominerals; dual-modality MRI; ferroptosis; photothermal therapy; protein corona

DOI:  https://doi.org/10.1021/acsnano.2c05917
Drug Deliv Transl Res. 2022 Dec 15.

A membrane-free microfluidic approach to mucus permeation for efficient differentiation of mucoadhesive and mucopermeating nanoparticulate systems.

Leah Wright, Anthony Wignall, Silver Jõemetsa, Paul Joyce, Clive A Prestidge.

  The gastrointestinal mucus barrier is a widely overlooked yet essential component of the intestinal epithelium, responsible for the body's protection against harmful pathogens and particulates. This, coupled with the increasing utilisation of biological molecules as therapeutics (e.g. monoclonal antibodies, RNA vaccines and synthetic proteins) and nanoparticle formulations for drug delivery, necessitates that we consider the additional absorption barrier that the mucus layer may pose. It is imperative that in vitro permeability methods can accurately model this barrier in addition to standardised cellular testing. In this study, a mucus-on-a-chip (MOAC) microfluidic device was engineered and developed to quantify the permeation kinetics of nanoparticles through a biorelevant synthetic mucus layer. Three equivalently sized nanoparticle systems, formulated from chitosan (CSNP), mesoporous silica (MSNP) and poly (lactic-co-glycolic) acid (PLGA-NP) were prepared to encompass various surface chemistries and nanostructures and were assessed for their mucopermeation within the MOAC. Utilising this device, the mucoadhesive behaviour of chitosan nanoparticles was clearly visualised, a phenomenon not often observed via standard permeation models. In contrast, MSNP and PLGA-NP displayed mucopermeation, with significant differences in permeation pattern due to specific mucus-nanoparticle binding. Further optimisation of the MOAC to include a more biorelevant mucus mimic resulted in 5.5-fold hindered PLGA-NP permeation compared to a mucin solution. Furthermore, tracking of PLGA-NP at a single nanoparticle resolution revealed rank-order correlations between particle diffusivity and MOAC permeation. This device, including utilisation of biosimilar mucus, provides a unique ability to quantify both mucoadhesion and mucopenetration of nano-formulations and elucidate mucus binding interactions on a microscopic scale.

Keywords:  Microfluidics; Mucoadhesion; Mucopermeation; Mucus; Nanoparticles; Organ-on-a-chip

DOI:  https://doi.org/10.1007/s13346-022-01274-8
Adv Mater. 2022 Dec 14. e2210208

Multimorphic Materials: Spatially Tailoring Mechanical Properties via Selective Initiation of Interpenetrating Polymer Networks.

Marshall J Allen, Hsu-Ming Lien, Nathaniel Prine, Carter Burns, Adrian Rylski, Xiaodan Gu, Lewis Cox, Filippo Mangolini, Benny D Freeman, Zachariah A Page.

  Access to multimaterial polymers with spatially localized properties and robust interfaces is anticipated to enable new capabilities in soft robotics, such as smooth actuation for advanced medical and manufacturing technologies. Here, orthogonal initiation is used to create interpenetrating polymer networks (IPNs) with spatial control over morphology and mechanical properties. Base catalyzes the formation of a stiff and strong polyurethane, while blue LEDs initiate the formation of a soft and elastic polyacrylate. IPN morphology is controlled by when the LED is turned "on", with large phase separation occurring for short time delays (∼1-2 minutes) and a mixed morphology for longer time delays (>5 minutes), which was supported by dynamic mechanical analysis, small angle X-ray scattering, and atomic force microscopy. Through tailoring morphology, tensile moduli and fracture toughness can be tuned across ∼1-2 orders of magnitude. Moreover, a simple spring model is used to explain the observed mechanical behavior. Photopatterning produces "multimorphic" materials, where morphology is spatially localized with fine precision (<100 μm), while maintaining a uniform chemical composition throughout to mitigate interfacial failure. The fabrication of hinges represents a possible use-case for multimorphic materials in soft robotics. This article is protected by copyright. All rights reserved.

Keywords:  interpenetrating networks; multimorphic; phase separation; photopatterning; photopolymerizations

DOI:  https://doi.org/10.1002/adma.202210208
Adv Drug Deliv Rev. 2022 Nov 26. pii: S0169-409X(22)00525-7. [Epub ahead of print]192 114635

Protein corona: Friend or foe? Co-opting serum proteins for nanoparticle delivery.

Woojun Kim, Nhu Ky Ly, Yanying He, Yongzhe Li, Zhongyue Yuan, Yoon Yeo.

  For systemically delivered nanoparticles to reach target tissues, they must first circulate long enough to reach the target and extravasate there. A challenge is that the particles end up engaging with serum proteins and undergo immune cell recognition and premature clearance. The serum protein binding, also known as protein corona formation, is difficult to prevent, even with artificial protection via "stealth" coating. Protein corona may be problematic as it can interfere with the interaction of targeting ligands with tissue-specific receptors and abrogate the so-called active targeting process, hence, the efficiency of drug delivery. However, recent studies show that serum protein binding to circulating nanoparticles may be actively exploited to enhance their downstream delivery. This review summarizes known issues of protein corona and traditional strategies to control the corona, such as avoiding or overriding its formation, as well as emerging efforts to enhance drug delivery to target organs via nanoparticles. It concludes with a discussion of prevailing challenges in exploiting protein corona for nanoparticle development.

Keywords:  Drug delivery; Nanoparticles; Protein corona; Stealth; Targeting

DOI:  https://doi.org/10.1016/j.addr.2022.114635
J Control Release. 2022 Dec 11. pii: S0168-3659(22)00836-7. [Epub ahead of print]

Protein-based nanocages for vaccine development.

Seong A Kim, Yeram Lee, Yeju Ko, Seohyun Kim, Gi Beom Kim, Na Kyeong Lee, Wonkyung Ahn, Nayeon Kim, Gi-Hoon Nam, Eun Jung Lee, In-San Kim.

  Protein nanocages have attracted considerable attention in various fields of nanomedicine due to their intrinsic properties, including biocompatibility, biodegradability, high structural stability, and ease of modification of their surfaces and inner cavities. In vaccine development, these protein nanocages are suited for efficient targeting to and retention in the lymph nodes and can enhance immunogenicity through various mechanisms, including excellent uptake by antigen-presenting cells and crosslinking with multiple B cell receptors. This review highlights the superiority of protein nanocages as antigen delivery carriers based on their physiological and immunological properties such as biodistribution, immunogenicity, stability, and multifunctionality. With a focus on design, we discuss the utilization and efficacy of protein nanocages such as virus-like particles, caged proteins, and artificial caged proteins against cancer and infectious diseases such as coronavirus disease 2019 (COVID-19). In addition, we summarize available knowledge on the protein nanocages that are currently used in clinical trials and provide a general outlook on conventional distribution techniques and hurdles faced, particularly for therapeutic cancer vaccines.

Keywords:  Cancer vaccine; Protein nanocage; SARS-CoV-2; Vaccine; Virus-like particle

DOI:  https://doi.org/10.1016/j.jconrel.2022.12.022
J Control Release. 2022 Dec 13. pii: S0168-3659(22)00847-1. [Epub ahead of print]

Elastin-like polypeptide-based micelles as a promising platform in nanomedicine.

Jolinde van Strien, Oscar Escalona-Rayo, Wim Jiskoot, Bram Slütter, Alexander Kros.

  New and improved nanomaterials are constantly being developed for biomedical purposes. Nanomaterials based on elastin-like polypeptides (ELPs) have increasingly shown potential over the past two decades. These polymers are artificial proteins of which the design is based on human tropoelastin. Due to this similarity, ELP-based nanomaterials are biodegradable and therefore well suited to drug delivery. The assembly of ELP molecules into nanoparticles spontaneously occurs at temperatures above a transition temperature (Tt). The ELP sequence influences both the Tt and the physicochemical properties of the assembled nanomaterial. Nanoparticles with desired properties can hence be designed by choosing the appropriate sequence. A promising class of ELP nanoparticles are micelles assembled from amphiphilic ELP diblock copolymers. Such micelles are generally uniform and well defined. Furthermore, site-specific attachment of cargo to the hydrophobic block results in micelles with the cargo shielded inside their core, while conjugation to the hydrophilic block causes the cargo to reside in the corona where it is available for interactions. Such control over particle design is one of the main contributing factors for the potential of ELP-based micelles as a drug delivery system. Additionally, the micelles are easily loaded with protein or peptide-based cargo by expressing it as a fusion protein. Small molecule drugs and other cargo types can be either covalently conjugated to ELP domains or physically entrapped inside the micelle core. This review aims to give an overview of ELP-based micelles and their applications in nanomedicine.

Keywords:  Drug delivery; Elastin-like polypeptides; Micelles; Nanomedicine; Thermo-responsiveness

DOI:  https://doi.org/10.1016/j.jconrel.2022.12.033
ACS Nano. 2022 Dec 16.

Highly Retina-Permeating and Long-Acting Resveratrol/Metformin Nanotherapeutics for Enhanced Treatment of Macular Degeneration.

Duc Dung Nguyen, Li-Jyuan Luo, Chia-Jung Yang, Jui-Yang Lai.

  The development of therapeutics for effective treatments of retinal diseases is significantly constrained by various biological barriers. We herein report a nanomedicine strategy to develop nanotherapeutics featured with not only high retinal permeability but also sustained bioactive delivery. Specifically, the nanotherapeutics are rationally designed via aminolysis of resveratrol-encapsulated polycaprolactone nanoparticles (R@PCL NPs), followed by the formation of amide linkages with carboxyl-terminated transacting activator of transcription cell penetrating peptide (T) and metformin (M). The R@PCL-T/M NP nanotherapeutics are demonstrated in vitro to possess persistent drug release profiles, good ocular biocompatibility, and potent bioactive activities for targeting prevailing risk factors associated with retinal diseases. In vivo studies indicate that single-dose intravitreal administration of the R@PCL-T/M NPs can effectively improve retinal permeability (∼15-fold increase), prevent loss of endogenous antioxidants, and suppress the growth of abnormal vessels in the retina with macular degeneration for 56 days. This high treatment efficacy can be ascribed to the enhanced retinal permeability of the nanotherapeutics in conjunction with the sustained pharmacological activity of the dual drugs (R and M) in the retinal pigment epithelial region. These findings show a great promise for the development of pharmacological nanoformulations capable of targeting the retina and thereby treating complex posterior segment diseases with improved efficacies.

Keywords:  Ocular nanomedicine; enhanced retinal permeation; macular degeneration; sustained drug release; therapeutic delivery system

DOI:  https://doi.org/10.1021/acsnano.2c05824