bims-drudre 2022-10-02 papers

bims-drudre

Biomed News

on Targeted drug delivery and programmed release mechanisms

Issue of 2022–10–02
twenty-two papers selected by
Ceren Kimna, Technical University of Munich

RoboCap: Robotic mucus-clearing capsule for enhanced drug delivery in the gastrointestinal tract.
A soft robotic sleeve mimicking the haemodynamics and biomechanics of left ventricular pressure overload and aortic stenosis.
Gastrointestinal tract drug delivery using algae motors embedded in a degradable capsule.
Lipid Nanoparticle (LNP) Chemistry Can Endow Unique In Vivo RNA Delivery Fates within the Liver That Alter Therapeutic Outcomes in a Cancer Model.
Spatial Transcriptomics-Based Identification of Molecular Markers for Nanomedicine Distribution in Tumor Tissue.
Macrophage-Membrane-Camouflaged Nonviral Gene Vectors for the Treatment of Multidrug-Resistant Bacterial Sepsis.
Molecularly engineered siRNA conjugates for tumor-targeted RNAi therapy.
Brain-targeted CRISPR/Cas9 nanomedicine for effective glioblastoma therapy.
Polarization of Tumor-Associated Macrophages Promoted by Vitamin C-Loaded Liposomes for Cancer Immunotherapy.
Deciphering microbial gene function using natural language processing.
Self-assembly of emulsion droplets through programmable folding.
3D Printed Hybrid Aerogel Gauzes Enable Highly Efficient Hemostasis.
Microencapsulation of Ionic Liquid by Interfacial Self-Assembly of Metal-Phenolic Network for Efficient Gastric Absorption of Oral Drug Delivery.
The reversion of DNA methylation-induced miRNA silence via biomimetic nanoparticles-mediated gene delivery for efficient lung adenocarcinoma therapy.
In situ small-angle X-ray scattering reveals strong condensation of DNA origami during silicification.
A Smart Nanoreactor Based on an O2-Economized Dual Energy Inhibition Strategy Armed with Dual Multi-stimuli-Responsive "Doorkeepers" for Enhanced CDT/PTT of Rheumatoid Arthritis.
Reshaping hypoxia and silencing CD73 via biomimetic gelatin nanotherapeutics to boost immunotherapy.
MicroRNA-7 regulates melanocortin circuits involved in mammalian energy homeostasis.
Photocatalytic glucose depletion and hydrogen generation for diabetic wound healing.
Robust Reversible Cross-Linking Strategy for Intracellular Protein Delivery with Excellent Serum Tolerance.
Magnetic Nanodroplets for Enhanced Deep Penetration of Solid Tumors and Simultaneous Magnetothermal-Sensitized Immunotherapy against Tumor Proliferation and Metastasis.
A Multifunctional Non-viral Vector for the Delivery of MTH1-targeted CRISPR/Cas9 System for Non-Small Cell Lung Cancer Therapy.

Sci Robot. 2022 Sep 28. 7(70): eabp9066

RoboCap: Robotic mucus-clearing capsule for enhanced drug delivery in the gastrointestinal tract.

Shriya S Srinivasan, Amro Alshareef, Alexandria V Hwang, Ziliang Kang, Johannes Kuosmanen, Keiko Ishida, Joshua Jenkins, Sabrina Liu, Wiam Abdalla Mohammed Madani, Jochen Lennerz, Alison Hayward, Josh Morimoto, Nina Fitzgerald, Robert Langer, Giovanni Traverso.

Oral drug delivery of proteins is limited by the degradative environment of the gastrointestinal tract and poor absorption, requiring parenteral administration of these drugs. Luminal mucus represents the initial steric and dynamic barrier to absorption. To overcome this barrier, we report the development of the RoboCap, an orally ingestible, robotic drug delivery capsule that locally clears the mucus layer, enhances luminal mixing, and topically deposits the drug payload in the small intestine to enhance drug absorption. RoboCap's mucus-clearing and churning movements are facilitated by an internal motor and by surface features that interact with small intestinal plicae circulares, villi, and mucus. Vancomycin (1.4 kilodaltons of glycopeptide) and insulin (5.8 kilodaltons of peptide) delivery mediated by RoboCap resulted in enhanced bioavailability 20- to 40-fold greater in ex vivo and in vivo swine models when compared with standard oral delivery (P < 0.05). Further, insulin delivery via the RoboCap resulted in therapeutic hypoglycemia, supporting its potential to facilitate oral delivery of drugs that are normally precluded by absorption limitations.

DOI: https://doi.org/10.1126/scirobotics.abp9066
Nat Biomed Eng. 2022 Sep 26.

A soft robotic sleeve mimicking the haemodynamics and biomechanics of left ventricular pressure overload and aortic stenosis.

Luca Rosalia, Caglar Ozturk, Jaume Coll-Font, Yiling Fan, Yasufumi Nagata, Manisha Singh, Debkalpa Goswami, Adam Mauskapf, Shi Chen, Robert A Eder, Efrat M Goffer, Jo H Kim, Salva Yurista, Benjamin P Bonner, Anna N Foster, Robert A Levine, Elazer R Edelman, Marcello Panagia, Jose L Guerrero, Ellen T Roche, Christopher T Nguyen.

Preclinical models of aortic stenosis can induce left ventricular pressure overload and coarsely control the severity of aortic constriction. However, they do not recapitulate the haemodynamics and flow patterns associated with the disease. Here we report the development of a customizable soft robotic aortic sleeve that can mimic the haemodynamics and biomechanics of aortic stenosis. By allowing for the adjustment of actuation patterns and blood-flow dynamics, the robotic sleeve recapitulates clinically relevant haemodynamics in a porcine model of aortic stenosis, as we show via in vivo echocardiography and catheterization studies, and a combination of in vitro and computational analyses. Using in vivo and in vitro magnetic resonance imaging, we also quantified the four-dimensional blood-flow velocity profiles associated with the disease and with bicommissural and unicommissural defects re-created by the robotic sleeve. The design of the sleeve, which can be adjusted on the basis of computed tomography data, allows for the design of patient-specific devices that may guide clinical decisions and improve the management and treatment of patients with aortic stenosis.

DOI: https://doi.org/10.1038/s41551-022-00937-8
Sci Robot. 2022 Sep 28. 7(70): eabo4160

Gastrointestinal tract drug delivery using algae motors embedded in a degradable capsule.

Fangyu Zhang, Zhengxing Li, Yaou Duan, Amal Abbas, Rodolfo Mundaca-Uribe, Lu Yin, Hao Luan, Weiwei Gao, Ronnie H Fang, Liangfang Zhang, Joseph Wang.

The use of micromotors for active drug delivery via oral administration has recently gained considerable interest. However, efficient motor-assisted delivery into the gastrointestinal (GI) tract remains challenging, owing to the short propulsion lifetime of currently used micromotor platforms. Here, we report on an efficient algae-based motor platform, which takes advantage of the fast and long-lasting swimming behavior of natural microalgae in intestinal fluid to prolong local retention within the GI tract. Fluorescent dye or cell membrane-coated nanoparticle functionalized algae motors were further embedded inside a pH-sensitive capsule to enhance delivery to the small intestines. In vitro, the algae motors displayed a constant motion behavior in simulated intestinal fluid after 12 hours of continuous operation. When orally administered in vivo into mice, the algae motors substantially improved GI distribution of the dye payload compared with traditional magnesium-based micromotors, which are limited by short propulsion lifetimes, and they also enhanced retention of a model chemotherapeutic payload in the GI tract compared with a passive nanoparticle formulation. Overall, combining the efficient motion and extended lifetime of natural algae-based motors with the protective capabilities of oral capsules results in a promising micromotor platform capable of achieving greatly improved cargo delivery in GI tissue for practical biomedical applications.

DOI: https://doi.org/10.1126/scirobotics.abo4160
Mol Pharm. 2022 Sep 26.

Lipid Nanoparticle (LNP) Chemistry Can Endow Unique In Vivo RNA Delivery Fates within the Liver That Alter Therapeutic Outcomes in a Cancer Model.

Lindsay T Johnson, Di Zhang, Kejin Zhou, Sang M Lee, Shuai Liu, Sean A Dilliard, Lukas Farbiak, Sumanta Chatterjee, Yu-Hsuan Lin, Daniel J Siegwart.

  Within the field of lipid nanoparticles (LNPs) for RNA delivery, the focus has been mainly placed on organ level delivery, which can mask cellular level effects consequential to therapeutic applications. Here, we studied a pair of LNPs with similar physical properties and discovered how the chemistry of the ionizable amino lipid can control the endogenous LNP identity, affecting cellular uptake in the liver and altering therapeutic outcomes in a model of liver cancer. Although most LNPs accumulate in the liver after intravenous administration (suggesting that liver delivery is straightforward), we observed an unexpected behavior when comparing two similar LNP formulations (5A2-SC8 and 3A5-SC14 LNPs) that resulted in distinct RNA delivery within the organ. Despite both LNPs possessing similar physical properties, ability to silence gene expression in vitro, strong accumulation within the liver, and a shared pKa of 6.5, only 5A2-SC8 LNPs were able to functionally deliver RNA to hepatocytes. Factor VII (FVII) activity was reduced by 87%, with 5A2-SC8 LNPs carrying FVII siRNA (siFVII), while 3A5-SC14 LNPs carrying siFVII produced baseline FVII activity levels comparable to the nontreatment control at a dosage of 0.5 mg/kg. Protein corona analysis indicated that 5A2-SC8 LNPs bind apolipoprotein E (ApoE), which can drive LDL-R receptor-mediated endocytosis in hepatocytes. In contrast, the surface of 3A5-SC14 LNPs was enriched in albumin but depleted in ApoE, which likely led to Kupffer cell delivery and detargeting of hepatocytes. In an aggressive MYC-driven liver cancer model relevant to hepatocytes, 5A2-SC8 LNPs carrying let-7g miRNA were able to significantly extend survival up to 121 days. Since disease targets exist in an organ- and cell-specific manner, the clinical development of RNA LNP therapeutics will require an improved understanding of LNP cellular tropism within organs. The results from our work illustrate the importance of understanding the cellular localization of RNA delivery and incorporating further checkpoints when choosing nanoparticles beyond biochemical and physical characterization, as small changes in the chemical composition of LNPs can have an impact on both the biofate of LNPs and therapeutic outcomes.

Keywords:  RNA delivery; cancer therapy; cell tropism; lipid nanoparticles; protein corona

DOI:  https://doi.org/10.1021/acs.molpharmaceut.2c00442
Small Methods. 2022 Sep 30. e2201091

Spatial Transcriptomics-Based Identification of Molecular Markers for Nanomedicine Distribution in Tumor Tissue.

Jeongbin Park, Jinyeong Choi, Jae Eun Lee, Hongyoon Choi, Hyung-Jun Im.

  The intratumoral accumulation of nanomedicine has been considered a passive process, referred to as the enhanced permeability and retention effect. Recent studies have suggested that the tumor uptake of nanomedicines follows an energy-dependent pathway rather than being a passive process. Herein, to explore the factor candidates that are associated with nanomedicine tumor uptake, a molecular marker identification platform is developed by integrating microscopic fluorescence images of a nanomedicine distribution with spatial transcriptomics information. When this approach is applied to PEGylated liposomes, molecular markers related to hypoxia, glycolysis, and apoptosis can be identified as being related to the intratumoral distribution of the nanomedicine. It is expected that the method can be applied to explain the distribution of a wide range of nanomedicines and that the data obtained from this analysis can enhance the precise utilization of nanomedicines.

Keywords:  apoptosis; enhanced permeability and retention; glycolysis; hypoxia; lipid nanoplatforms; molecular markers; spatial transcriptomics

DOI:  https://doi.org/10.1002/smtd.202201091
Nano Lett. 2022 Sep 28.

Macrophage-Membrane-Camouflaged Nonviral Gene Vectors for the Treatment of Multidrug-Resistant Bacterial Sepsis.

Hongmei Cao, Yang Gao, Haixue Jia, Liping Zhang, Jinjian Liu, Ganen Mu, Han Gui, Yuebing Wang, Cuihong Yang, Jianfeng Liu.

  Sepsis is a life-threatening disease caused by systemic bacterial infections, with high morbidity and mortality worldwide. As the standard treatment for sepsis, antibiotic therapy faces the challenge of impaired macrophages and drug-resistant bacteria. In this study, we developed a membrane-camouflaged metal-organic framework (MOF) system for plasmid DNA (pDNA) delivery to combat sepsis. The antimicrobial gene LL37 was efficiently encapsulated in the pH-sensitive MOF, and the nanoparticles were decorated with macrophage membranes in a compatible manner. Macrophage membrane coating allows targeted delivery of LL37 to macrophages and creates macrophage factories for the continuous generation of antimicrobial peptides. Compared to naked nanoparticles, primary bone marrow mesenchymal macrophage membrane-modified nanoparticles greatly improved the survival rate of immunodeficient septic mice through the synergistic effect of efficient gene therapy and inflammatory cytokine sequestration. This study demonstrates an effective membrane biomimetic strategy for efficiently delivering pDNA, offering an excellent option for overcoming sepsis.

Keywords:  biomimetic nanoparticles; gene therapy; multidrug-resistant bacterial sepsis; pDNA intracellular delivery; primary bone marrow mesenchymal macrophage membrane

DOI:  https://doi.org/10.1021/acs.nanolett.2c02560
J Control Release. 2022 Sep 21. pii: S0168-3659(22)00635-6. [Epub ahead of print]

Molecularly engineered siRNA conjugates for tumor-targeted RNAi therapy.

Jong Won Lee, Jiwon Choi, Yeonho Choi, Kwangmeyung Kim, Yoosoo Yang, Sun Hwa Kim, Hong Yeol Yoon, Ick Chan Kwon.

  RNA interference (RNAi) is a major cellular mechanism regulating gene expression in which short double-stranded RNA molecules called small interfering RNA (siRNA) mediate sequence-specific mRNA degradation. RNAi technology has recently emerged as a promising therapeutic platform for the effective treatment of various diseases caused by inappropriate gene activity, such as cancer. However, the clinical translation of siRNA therapeutics has been hampered by the major hurdles associated with biological instability and limited delivery efficiency. Based on the various efforts, recent siRNA delivery strategies using cationic lipids and polymers allowed to enhance pharmacokinetics and delivery efficiency, resulting in potent and liver-targeted RNAi therapy. However, non-specific protein adsorption, high liver accumulation, and severe toxicity of cationic nanocarriers still limit the possibility of transfer of siRNA therapeutics from the laboratory to the clinic. One of the promising delivery strategies to overcome the limitations of siRNA therapeutics is carrier-free bioconjugation which is chemically modified and connected with biocompatible molecules such as lipids, peptides, antibodies, aptamers, and polymers. These molecularly engineered siRNA conjugates can be utilized for RNAi delivery to tissues beyond the liver, providing opportunities for clinical translation. This review focused on introducing the recent progress in molecularly engineered siRNA conjugates and their applications toward overcoming the limitations of siRNA for tumor-targeted delivery and therapy.

Keywords:  Gene delivery; RNA interference; Small interfering RNA; cancer therapy; siRNA conjugates

DOI:  https://doi.org/10.1016/j.jconrel.2022.09.040
J Control Release. 2022 Sep 26. pii: S0168-3659(22)00641-1. [Epub ahead of print]

Brain-targeted CRISPR/Cas9 nanomedicine for effective glioblastoma therapy.

Weimin Ruan, Mingzhu Jiao, Sen Xu, Muhammad Ismail, Xuan Xie, Yang An, Haixing Guo, Rongjun Qian, Bingyang Shi, Meng Zheng.

  CRISPR/Cas9 gene-editing technology shows great potential for treating a variety of diseases, such as glioblastoma multiforme (GBM). However, CRISPR components suffer from inherent delivery challenges, such as poor in vivo stability of Cas9 protein and gRNA, low blood-brain barrier (BBB) permeability and non-specific tissue or cell targeting. These defects have limited the application of Cas9/gRNA ribonucleoprotein (RNP) complexes for GBM therapy. Here, we developed a brain-targeted CRISPR/Cas9 based nanomedicine by fabricating an angiopep-2 decorated, guanidinium and fluorine functionalized polymeric nanoparticle with loading Cas9/gRNA RNP for the treatment of GBM. The guanidinium and fluorine domains of our polymeric nanoparticles were both capable of interacting with Cas9/gRNA RNP to stabilize it in blood circulation, without impairing its activity. Moreover, by leveraging angiopep-2 peptide functionality, the RNP nanoparticles efficiently crossed the BBB and accumulated in brain tumors. In U87MG cells, we achieved approximately 32% gene knockout and 67% protein reduction in the targeted proto-oncogene polo-like kinase 1 (PLK1). This was sufficient to suppress tumor growth and significantly improved the median survival time of mice bearing orthotopic glioblastoma to 40 days, while inducing negligible side or off-target effects. These results suggest that the developed brain-targeted CRISPR/Cas9 based nanomedicine shows promise for effective human glioblastoma gene therapy.

Keywords:  Brain-targeting; CRISPR/Cas9; Gene-editing; Glioblastoma; Nanoparticle

DOI:  https://doi.org/10.1016/j.jconrel.2022.09.046
ACS Nano. 2022 Sep 27.

Polarization of Tumor-Associated Macrophages Promoted by Vitamin C-Loaded Liposomes for Cancer Immunotherapy.

Zhaoyu Ma, Mingkun Yang, Mohamed Frahat Foda, Kai Zhang, Shuting Li, Huageng Liang, Yanli Zhao, Heyou Han.

  While checkpoint blockade immunotherapy as a promising clinical modality has revolutionized cancer treatment, it is of benefit to only a subset of patients because of the tumor immunosuppressive microenvironment. Herein, we report that the specified delivery of vitamin C at the tumor site by responsive lipid nanoparticles can efficiently induce oxidative toxicity and the polarization of M1 macrophages, promoting the infiltration of activating cytotoxic T lymphocytes in the tumor microenvironment for intensive immune checkpoint blocking therapy. Both in vitro and in vivo assays demonstrate successful vitamin C-induced polarization of M2 macrophages to M1 macrophages. In vivo transcriptome analysis also reveals the activation mechanism of vitamin C immunity. More importantly, the combination approach displays much better immune response and immune process within the tumor microenvironment than clinical programmed cell death ligand 1 (Anti-PD-L1) alone. This work provides a powerful therapeutic application of vitamin C to amplify Anti-PD-L1 immunotherapy in cancer treatment, which brings hope to patients with clinically insensitive immunity.

Keywords:  Anti-PD-L1; checkpoint blockade immunotherapy; macrophage polarization; tumor immunosuppressive microenvironment; vitamin C

DOI:  https://doi.org/10.1021/acsnano.2c08446
Nat Commun. 2022 Sep 29. 13(1): 5731

Deciphering microbial gene function using natural language processing.

Danielle Miller, Adi Stern, David Burstein.

Revealing the function of uncharacterized genes is a fundamental challenge in an era of ever-increasing volumes of sequencing data. Here, we present a concept for tackling this challenge using deep learning methodologies adopted from natural language processing (NLP). We repurpose NLP algorithms to model "gene semantics" based on a biological corpus of more than 360 million microbial genes within their genomic context. We use the language models to predict functional categories for 56,617 genes and find that out of 1369 genes associated with recently discovered defense systems, 98% are inferred correctly. We then systematically evaluate the "discovery potential" of different functional categories, pinpointing those with the most genes yet to be characterized. Finally, we demonstrate our method's ability to discover systems associated with microbial interaction and defense. Our results highlight that combining microbial genomics and language models is a promising avenue for revealing gene functions in microbes.

DOI: https://doi.org/10.1038/s41467-022-33397-4
Nature. 2022 Sep 28.

Self-assembly of emulsion droplets through programmable folding.

Angus McMullen, Maitane Muñoz Basagoiti, Zorana Zeravcic, Jasna Brujic.

In the realm of particle self-assembly, it is possible to reliably construct nearly arbitrary structures if all the pieces are distinct1-3, but systems with fewer flavours of building blocks have so far been limited to the assembly of exotic crystals4-6. Here we introduce a minimal model system of colloidal droplet chains7, with programmable DNA interactions that guide their downhill folding into specific geometries. Droplets are observed in real space and time, unravelling the rules of folding. Combining experiments, simulations and theory, we show that controlling the order in which interactions are switched on directs folding into unique structures, which we call colloidal foldamers8. The simplest alternating sequences (ABAB...) of up to 13 droplets yield 11 foldamers in two dimensions and one in three dimensions. Optimizing the droplet sequence and adding an extra flavour uniquely encodes more than half of the 619 possible two-dimensional geometries. Foldamers consisting of at least 13 droplets exhibit open structures with holes, offering porous design. Numerical simulations show that foldamers can further interact to make complex supracolloidal architectures, such as dimers, ribbons and mosaics. Our results are independent of the dynamics and therefore apply to polymeric materials with hierarchical interactions on all length scales, from organic molecules all the way to Rubik's Snakes. This toolbox enables the encoding of large-scale design into sequences of short polymers, placing folding at the forefront of materials self-assembly.

DOI: https://doi.org/10.1038/s41586-022-05198-8
Adv Healthc Mater. 2022 Sep 27. e2201591

3D Printed Hybrid Aerogel Gauzes Enable Highly Efficient Hemostasis.

Xiaoxu Yang, Nan Shi, Jian Liu, Qingqing Cheng, Guangyong Li, Jing Lyu, Fengguo Ma, Xuetong Zhang.

  Hemostatic materials have played a significant role in mitigating traumatic injury by controlling the bleeding, however the fabrication of desirable material's structure to enhance the accumulation of blood cells and platelets for highly efficient hemostasis is still a great challenge. In this work, directed assembly of poly(vinyl alcohol) (PVA) macromolecules covering the rigid Kevlar nanofiber (KNF) network during 3D printing process, is utilized to fabricate hydrophilic, biocompatible, and mechanically stable KNF-PVA aerogel filaments for effective enriching blood components by fast water absorption. As such, KNF-PVA aerogel gauzes demonstrate remarkable water permeability (338 mL cm-2 s-1 bar-1 ), water absorption speed (as high as 9.64 g g-1 min-1 ) and capacity (more than 10 times of self-weight), and ability to enrich micron-sized particles when contacting aqueous solution. All these properties favor efficient hemostasis and the resulting KNF-PVA aerogel gauzes significantly outperform the commercial product Quikclot Gauze (QCG, Z-Medica) during in-vivo experiments with the rat liver laceration model, reducing the hemostasis time by half (60 ± 4 s) and the blood loss by two thirds (0.07 ± 0.01 g). These results demonstrate a robust strategy to design various aerogel gauzes for hemostasis applications. This article is protected by copyright. All rights reserved.

Keywords:  3D printing; Aerogel gauze; Kevlar nanofiber; hemostatic materials; poly(vinyl alcohol)

DOI:  https://doi.org/10.1002/adhm.202201591
ACS Appl Mater Interfaces. 2022 Sep 29.

Microencapsulation of Ionic Liquid by Interfacial Self-Assembly of Metal-Phenolic Network for Efficient Gastric Absorption of Oral Drug Delivery.

Lanbo Shen, Yaping Zhang, Junkun Feng, Wenxiu Xu, Yi Chen, Kai Li, Xiaoru Yang, Yajun Zhao, Shaohua Ge, Jianhua Li.

  Improving bioavailability of orally delivered drugs is still challenging, as conventional drug delivery systems suffer from non-specific drug delivery in the gastrointestinal (GI) tract and limited drug absorption efficiency. Gastric drug delivery is even more difficult due to the harsh microenvironment, short retention time, and physiologic barriers in the stomach. Here, an oral drug delivery microcapsule system was developed for gastric drug delivery, which consists of ionic liquid (IL) as the inner carrier and metal-phenolic network (MPN) as the microcapsule shell. The IL@MPN microcapsules are prepared by interfacial self-assembly of FeIII and quercetin at the interface of hydrophobic IL ([EMIM][NTf2]) and water. The formation of MPN shell could improve the stability of IL droplets in water and endow the system with pH-response drug release properties, while the encapsulated IL core could efficiently load the drug and enhance the drug tissue permeability. The IL@MPN microcapsules showed enhanced drug absorption in the stomach after oral administration in a rat model, where the microcapsules are disassembled in gastric acid, and the released IL could reduce the viscosity of mucus gel and increase the drug transport rate across endothelial cells. This work presents a simple yet efficient strategy for oral drug delivery to the stomach. Given the diversity and versatility of both MPN and IL, the proposed self-assembled microcapsules could expand the toolbox of drug delivery systems with enhanced oral drug bioavailability.

Keywords:  interfacial self-assembly; ionic liquids; metal-phenolic network; microcapsule; oral drug delivery

DOI:  https://doi.org/10.1021/acsami.2c15599
Mol Cancer. 2022 Sep 28. 21(1): 186

The reversion of DNA methylation-induced miRNA silence via biomimetic nanoparticles-mediated gene delivery for efficient lung adenocarcinoma therapy.

Lu Liang, Huiyu Cen, Jionghua Huang, Aiping Qin, Wenyan Xu, Siran Wang, Zhijun Chen, Lin Tan, Qiqi Zhang, Xiyong Yu, Xin Yang, Lingmin Zhang.

   BACKGROUND: Lung cancer is one of the fatal cancers worldwide, and over 60% of patients are lung adenocarcinoma (LUAD). Our clinical data demonstrated that DNA methylation of the promoter region of miR-126-3p was upregulated, which led to the decreased expression of miR-126-3p in 67 cases of lung cancer tissues, implying that miR-126-3p acted as a tumor suppressor. Transduction of miR-126-3p is a potential therapeutic strategy for treating LUAD, yet the physiological environment and properties of miRNA challenge current transduction approaches.
METHODS: We evaluated the expression of miR-126-3p in 67 pairs of lung cancer tissues and the corresponding adjacent non-tumorous tissues by Reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The relationship between the overall survival of lung cancer patients and miR-126-3p was analyzed by the Cancer Genome Atlas cohort database (Oncolnc, http://www.oncolnc.org ). We analyzed DNA methylation Methylation-specific PCR (MSP) analysis. To determine whether ADAM9 is the direct target of miR-126-3p, we performed the 3'-UTR luciferase reporter assay. The protein levels in the cells or tissues were evaluated with western blotting (WB) analysis. The biodistribution of nanoparticles were monitored by in vivo tracking system.
RESULTS: We describe the development of novel stealth and matrix metalloproteinase 2 (MMP2)-activated biomimetic nanoparticles, which are constructed using MMP2-responsive peptides to bind the miR-126-3p (known as MAIN), and further camouflaged with red blood cell (RBC) membranes (hence named REMAIN). REMAIN was able to effectively transduce miRNA into lung cancer cells and release them via MMP2 responsiveness. Additionally, REMAIN possessed the advantages of the natural RBC membrane, including extended circulation time, lower toxicity, better biocompatibility, and immune escape. Moreover, in vitro and in vivo results demonstrated that REMAIN effectively induced apoptosis of lung cancer cells and inhibited LUAD development and progression by targeting ADAM9.
CONCLUSION: The novel style of stealth and MMP2-activated biomimetic nanoparticles show great potential in miRNA delivery.

Keywords:  Biomimetic nanoparticles; DNA methylation; Lung adenocarcinoma; MMP2; MicroRNA

DOI:  https://doi.org/10.1186/s12943-022-01651-4
Nat Commun. 2022 Sep 27. 13(1): 5668

In situ small-angle X-ray scattering reveals strong condensation of DNA origami during silicification.

Martina F Ober, Anna Baptist, Lea Wassermann, Amelie Heuer-Jungemann, Bert Nickel.

Silicification of DNA origami structures increases their stability and provides chemical protection. Yet, it is unclear whether the whole DNA framework is embedded or if silica just forms an outer shell and how silicification affects the origami's internal structure. Employing in situ small-angle X-ray scattering (SAXS), we show that addition of silica precursors induces substantial condensation of the DNA origami at early reaction times by almost 10 %. Subsequently, the overall size of the silicified DNA origami increases again due to increasing silica deposition. We further identify the SAXS Porod invariant as a reliable, model-free parameter for the evaluation of the amount of silica formation at a given time. Contrast matching of the DNA double helix Lorentzian peak reveals silica growth also inside the origami. The less polar silica forming within the origami structure, replacing more than 40 % of the internal hydration water, causes a hydrophobic effect: condensation. DNA origami objects with flat surfaces show a strong tendency towards aggregation during silicification, presumably driven by the same entropic forces causing condensation. Maximally condensed origami displayed thermal stability up to 60 °C. Our studies provide insights into the silicification reaction allowing for the formulation of optimized reaction protocols.

DOI: https://doi.org/10.1038/s41467-022-33083-5
ACS Nano. 2022 Sep 26.

A Smart Nanoreactor Based on an O2-Economized Dual Energy Inhibition Strategy Armed with Dual Multi-stimuli-Responsive "Doorkeepers" for Enhanced CDT/PTT of Rheumatoid Arthritis.

Shang Qiu, Xiunan Wu, Zheng Li, Xinyu Xu, Juan Wang, Yawei Du, Wenzhen Pan, Ruqi Huang, Yafei Wu, Zhi Yang, Qi Zhou, Bing Zhou, Xuren Gao, Yaozeng Xu, Wenguo Cui, Fenglei Gao, Dechun Geng.

  Activated fibroblast-like synovial (FLS) cells are regarded as an important target for rheumatoid arthritis (RA) treatment via starvation therapy mediated by glucose oxidase (GOx). However, the hypoxic RA-FLS environment greatly reduces the oxidation process of glucose and leads to a poor therapeutic effect of the GOx-based starvation therapy. In this work, we designed a hollow mesoporous copper sulfide nanoparticles (CuS NPs)-based smart GOx/atovaquone (ATO) codelivery system (named as V-HAGC) targeting RA-FLS cells to realize a O2-economized dual energy inhibition strategy to solve the limitation of GOx-based starvation therapy. V-HAGC armed with dual multi-stimuli-responsive "doorkeepers" can guard drugs intelligently. Once under the stimulation of photothermal and acidic conditions at the targeted area, the dual intelligent responsive "doors" would orderly open to realize the controllable release of drugs. Besides, the efficacy of V-HAGC would be much improved by the additional chemodynamic therapy (CDT) and photothermal therapy (PTT) stimulated by CuS NPs. Meanwhile, the upregulated H2O2 and acid levels by starvation therapy would promote the Fenton-like reaction of CuS NPs under O2-economized dual energy inhibition, which could enhance the PTT and CDT efficacy as well. In vitro and in vivo evaluations revealed V-HAGC with much improved efficacy of this combination therapy for RA. In general, the smart V-HAGC based on the O2-economized dual energy inhibition strategy combined with enhanced CDT and PTT has the potential to be an alternative methodology in the treatment of RA.

Keywords:  O2-economy; chemodynamic therapy; dual energy suppression; rheumatoid arthritis; starvation therapy

DOI:  https://doi.org/10.1021/acsnano.2c07338
J Control Release. 2022 Sep 22. pii: S0168-3659(22)00624-1. [Epub ahead of print]351 255-271

Reshaping hypoxia and silencing CD73 via biomimetic gelatin nanotherapeutics to boost immunotherapy.

Cong-Shan Yuan, Zhuang Teng, Shuang Yang, Zheng He, Ling-Yang Meng, Xi-Guang Chen, Ya Liu.

  The ubiquitous hypoxic microenvironment at the tumor site helps to regulate hypoxic inducible factor (HIF-1α), up-regulate downstream CD73-adenosine (CD73-ADO) pathways, and further result in effector T cell function exhaustion, which is regarded as a crucial adverse factor in the poor clinical efficacy of immune checkpoint blockade therapy (ICB). How to reshape hypoxic microenvironment and silence CD73 remains a huge challenge to improve ICB therapeutic outcomes. In this study, cancer cell membrane-camouflaged gelatin nanoparticles (CSG@B16F10) were designed to co-deliver oxygen-generating agent catalase (CAT) and CD73siRNA, thus enhancing tumor oxygenation and alleviating CD73-ADO pathway-mediated T cell immunosuppression. The fabricated biomimetic nanoparticles could efficiently achieve immune evading and homologous targeting by virtue of the retention of cancer cell membrane protein. Matrix metalloproteinases (MMP)-responsive gelatin nanoparticles were gradually disintegrated to accelerate the release of payloads. Rapidly released CAT was found to relieve tumor hypoxia by generating endogenous oxygen, while CD73siRNA effectively silenced target gene, synergically inhibiting CD73 protein expression and facilitating T-cell-specific immunity. Upon introduction of CSG@B16F10 in melanoma-bearing mice, PD-L1 checkpoint blockade achieved optimal tumor suppression (∼83%). The enhanced immune efficacy was mainly manifested by enhanced cytotoxic T cell (CTL), reduced regulatory T cells (Tregs), and increased anti-tumor cytokine secretion. This work presents a new paradigm for the ideal design of biomimetic nanoplatforms and the synergistic treatment of hypoxia alleviation and CD73 silence, greatly promising for enhancing clinical immune potency of PD-1/PD-L1 immune checkpoint blockade.

Keywords:  CD73 silence; Gelatin nanoparticle; Hypoxia alleviation; Immunotherapy; cancer cytomembrane camouflage

DOI:  https://doi.org/10.1016/j.jconrel.2022.09.029
Nat Commun. 2022 Sep 29. 13(1): 5733

MicroRNA-7 regulates melanocortin circuits involved in mammalian energy homeostasis.

Mary P LaPierre, Katherine Lawler, Svenja Godbersen, I Sadaf Farooqi, Markus Stoffel.

MicroRNAs (miRNAs) modulate physiological responses by repressing the expression of gene networks. We found that global deletion of microRNA-7 (miR-7), the most enriched miRNA in the hypothalamus, causes obesity in mice. Targeted deletion of miR-7 in Single-minded homolog 1 (Sim1) neurons, a critical component of the hypothalamic melanocortin pathway, causes hyperphagia, obesity and increased linear growth, mirroring Sim1 and Melanocortin-4 receptor (MC4R) haplo-insufficiency in mice and humans. We identified Snca (α-Synuclein) and Igsf8 (Immunoglobulin Superfamily Member 8) as miR-7 target genes that act in Sim1 neurons to regulate body weight and endocrine axes. In humans, MIR-7-1 is located in the last intron of HNRNPK, whose promoter drives the expression of both genes. Genetic variants at the HNRNPK locus that reduce its expression are associated with increased height and truncal fat mass. These findings demonstrate that miR-7 suppresses gene networks involved in the hypothalamic melanocortin pathway to regulate mammalian energy homeostasis.

DOI: https://doi.org/10.1038/s41467-022-33367-w
Nat Commun. 2022 Sep 27. 13(1): 5684

Photocatalytic glucose depletion and hydrogen generation for diabetic wound healing.

Shengqiang Chen, Yanxia Zhu, Qingqing Xu, Qi Jiang, Danyang Chen, Ting Chen, Xishen Xu, Zhaokui Jin, Qianjun He.

High-glucose microenvironment in the diabetic foot ulcer (DFU) causes excessive glycation and induces chronic inflammation, leading to the difficulty of DFU healing. Hydrogen-rich water bath can promote the healing of DFU in clinic by virtue of the anti-inflammatory effect of hydrogen molecules, but the long-term daily soaking counts against the formation of a scab and cannot change the high-glucose microenvironment, limiting the outcome of DFU therapy. In this work, photocatalytic therapy of diabetic wound is proposed for sustainable hydrogen generation and local glucose depletion by utilizing glucose in the high-glucose microenvironment as a sacrificial agent. Hydrogen-incorporated titanium oxide nanorods are developed to realize efficient visible light (VIS)-responsive photocatalysis for glucose depletion and hydrogen generation, achieving a high efficacy of diabetic wound healing. Mechanistically, local glucose depletion and hydrogen generation jointly attenuate the apoptosis of skin cells and promote their proliferation and migration by inhibiting the synthesis of advanced glycation end products and the expression of their receptors, respectively. The proposed VIS-photocatalytic strategy provides a solution for facile, safe and efficient treatment of DFU.

DOI: https://doi.org/10.1038/s41467-022-33475-7
Nano Lett. 2022 Sep 29.

Robust Reversible Cross-Linking Strategy for Intracellular Protein Delivery with Excellent Serum Tolerance.

Song Zhang, Echuan Tan, Ruijue Wang, Peng Gao, Hui Wang, Yiyun Cheng.

  Intracellular protein delivery has attracted increasing attentions in biomedical applications. However, current delivery systems usually have poor serum stability due to the competitive binding of serum proteins to the polymers during delivery. Here, we report a reversible cross-linking strategy to improve the serum stability of polymers for robust intracellular protein delivery. In the proposed delivery system, nanoparticles are assembled by cargo proteins and cationic polymers and further stabilized by a glutathione-cleavable and traceless cross-linker. The cross-linked nanoparticles show high stability and efficient cell internalization in serum containing medium and can release the cargo proteins in response to intracellular glutathione and acidic pH in a traceless manner. The generality and versatility of the proposed strategy were demonstrated on different types of cationic polymers, cargo proteins, as well as cell lines. The study provides a facile and efficient method for improving the serum tolerance of cationic polymers in intracellular protein delivery.

Keywords:  intracellular protein delivery; nanoparticles; polymers; reversible cross-linking; serum tolerance

DOI:  https://doi.org/10.1021/acs.nanolett.2c02948
Adv Healthc Mater. 2022 Sep 27. e2201399

Magnetic Nanodroplets for Enhanced Deep Penetration of Solid Tumors and Simultaneous Magnetothermal-Sensitized Immunotherapy against Tumor Proliferation and Metastasis.

Junrui Wang, Wenli Zhang, Zhuoyan Xie, Xingyue Wang, Ying Luo, Weixi Jiang, Yun Liu, Zhigang Wang, Haitao Ran, Weixiang Song, Dajing Guo.

  The central cells of solid tumors are more proliferative and metastatic than the marginal cells. Therefore, more intelligent strategies for targeting cells with deep spatial distributions in solid tumors remain to be explored. In this work, a biocompatible nanotheranostic agent with a lipid membrane-coated, Fe3 O4 and perfluoropentane (PFP)-loaded, cRGD peptide (specifically targeting the integrin αvβ3 receptor)-grafted, magnetic nanodroplets (MNDs) was developed. The MNDs exhibited excellent magnetothermal conversion and controllable magnetic hyperthermia (MHT) through alternating magnetic field regulation. Furthermore, MHT-mediated magnetic droplet vaporization (MDV) induced the expansion of the MNDs to transform them into ultrasonic microbubbles, increasing the permeability of tissue and the cell membrane via the ultrasound-targeted microbubble destruction (UTMD) technique and thereby promoting the deep penetration of MNDs in solid tumors. More importantly, MHT not only caused apoptotic damage by downregulating the expression of the HSP70, cyclin D1, and Bcl-2 proteins in tumor cells but also improved the response rate to T-cell-related immunotherapy by upregulating PD-L1 expression in tumor cells, thus inhibiting the growth of both primary and metastatic tumors. Overall, this work introduces a distinct application of nanoultrasonic biomedicine in cancer therapy and provides an attractive immunotherapy strategy for preventing the proliferation and metastasis of deeply distributed cells in solid tumors. This article is protected by copyright. All rights reserved.

Keywords:  immunotherapy; magnetic droplet vaporization, magnetic fluid hyperthermia; nanoultrasonic biomedicine; solid tumor

DOI:  https://doi.org/10.1002/adhm.202201399
Acta Biomater. 2022 Sep 23. pii: S1742-7061(22)00615-8. [Epub ahead of print]

A Multifunctional Non-viral Vector for the Delivery of MTH1-targeted CRISPR/Cas9 System for Non-Small Cell Lung Cancer Therapy.

Yu Wang, Yan Tang, Xiao-Mei Zhao, Gui Huang, Jin-Hong Gong, Shu-di Yang, Hui Li, Wen-Jun Wan, Chang-Hao Jia, Gang Chen, Xue-Nong Zhang.

  Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system adapted from bacteria is a programmable nuclease-based genome editing tool. The long-lasting effect of gene silencing or correction is beneficial in cancer treatment. Considering the need to broaden the practical application of this technology, highly efficient non-viral vectors are urgently required. We prepared a multifunctional non-viral vector that could actively target tumor cells and deliver CRISPR/Cas9 plasmids into nuclei of cancer cells. Protamine sulfate (PS) which contains nuclear localization sequence was utilized to condense plasmid DNA and facilitate nuclei-targeted delivery. Liposome-coated protein/DNA complex avoided the degradation of nuclease in blood circulation. The obtained PS@Lip/pCas9 was further modified with distearoyl phosphoethanolamine-polyethylene glycol-hyaluronic acid (HA) to endow the vector ability to actively target tumor cell. Results suggested that PS@HA-Lip could deliver CRISPR/Cas9 plasmids into nuclei of tumor cells and induce genome editing effect. With the disruption of MTH1 (mutT homolog1) gene, the growth of non-small cell lung cancer was inhibited. Moreover, cell apoptosis in tumor tissue was promoted, and liver metastasis of non-small cell lung cancer (NSCLC) was reduced. Our study has provided a therapeutic strategy targeting MTH1 gene for NSCLC therapy. STATEMENT OF SIGNIFICANCE: CRISPR/Cas9 as a powerful tool for genome editing has drawn much attention. The long-lasting effect possesses unique advantage in cancer treatment. Non-viral vectors have high loading capacity, high safety and low immunogenicity, playing an important role in CRISPR/Cas9 delivery. In our study, a multifunctional non-viral vector for the efficient delivery of CRISPR/Cas9 plasmid was constructed. With the active targeting ligand and nuclei-targeting component, the cargo was efficiently delivered into cell nuclei and exerted genome editing effect. By using this vector, we successfully inhibited the growth and induced the apoptosis of non-small cell lung cancer by disrupting MTH1 expression with good safety. Our work provided an efficient non-vial vector for CRISPR/Cas9 delivery and explored the possibility for cancer treatment.

Keywords:  CRISPR/Cas9; non-viral vector; targeted gene therapy; targeted nano delivery; tumor

DOI:  https://doi.org/10.1016/j.actbio.2022.09.046