bims-drudre 2021-01-24 papers

bims-drudre

Biomed News

on Targeted drug delivery and programmed release mechanisms

Issue of 2021–01–24
twenty-two papers selected by
Ceren Kimna, Technical University of Munich

Evolution of antibody immunity to SARS-CoV-2.
Distinct metabolic programs established in the thymus control effector functions of γδ T cell subsets in tumor microenvironments.
Host succinate is an activation signal for Salmonella virulence during intracellular infection.
Regenerating Antithrombotic Surfaces through Nucleic Acid Displacement.
Ionic shape-morphing microrobotic end-effectors for environmentally adaptive targeting, releasing, and sampling.
Design of SARS-CoV-2 hFc-Conjugated Receptor-Binding Domain mRNA Vaccine Delivered via Lipid Nanoparticles.
Immunogenic Hybrid Nanovesicles of Liposomes and Tumor-Derived Nanovesicles for Cancer Immunochemotherapy.
Biomimetic Design of Mitochondria-Targeted Hybrid Nanozymes as Superoxide Scavengers.
Metabolic adaptation of acute lymphoblastic leukemia to the central nervous system microenvironment is dependent on Stearoyl CoA desaturase.
Integration of Multitargeted Polymer-Based Contrast Agents with Photoacoustic Computed Tomography: An Imaging Technique to Visualize Breast Cancer Intratumor Heterogeneity.
Developmental cell programs are co-opted in inflammatory skin disease.
Low-dose decitabine priming endows CAR T cells with enhanced and persistent antitumour potential via epigenetic reprogramming.
Recent Advances of DNA Nanostructure-Based Cell Membrane Engineering.
Core transcription regulatory circuitry orchestrates corneal epithelial homeostasis.
A Biomimetic Drug Delivery System by Integrating Grapefruit Extracellular Vesicles and Doxorubicin-Loaded Heparin-Based Nanoparticles for Glioma Therapy.
Rapid Development of SARS-CoV-2 Spike Protein Receptor-Binding Domain Self-Assembled Nanoparticle Vaccine Candidates.
CRISPR technologies and the search for the PAM-free nuclease.
Adeno-associated virus-mediated delivery of anti-miR-199a tough decoys attenuates cardiac hypertrophy by targeting PGC-1alpha.
Boosting anti-PD-1 therapy with metformin-loaded macrophage-derived microparticles.
Oral Administration of Starting Materials for In Vivo Synthesis of Antibacterial Gold Nanoparticles for Curing Remote Infections.
Hydrogel microspheres for spatiotemporally controlled delivery of RNA and silencing gene expression within scaffold-free tissue engineered constructs: Spatiotemporally controlled gene expression within MSC aggregates.
How Functionalized Surfaces Can Inhibit Bacterial Adhesion and Viability.

Nature. 2021 Jan 18.

Evolution of antibody immunity to SARS-CoV-2.

Christian Gaebler, Zijun Wang, Julio C C Lorenzi, Frauke Muecksch, Shlomo Finkin, Minami Tokuyama, Alice Cho, Mila Jankovic, Dennis Schaefer-Babajew, Thiago Y Oliveira, Melissa Cipolla, Charlotte Viant, Christopher O Barnes, Yaron Bram, Gaëlle Breton, Thomas Hägglöf, Pilar Mendoza, Arlene Hurley, Martina Turroja, Kristie Gordon, Katrina G Millard, Victor Ramos, Fabian Schmidt, Yiska Weisblum, Divya Jha, Michael Tankelevich, Gustavo Martinez-Delgado, Jim Yee, Roshni Patel, Juan Dizon, Cecille Unson-O'Brien, Irina Shimeliovich, Davide F Robbiani, Zhen Zhao, Anna Gazumyan, Robert E Schwartz, Theodora Hatziioannou, Pamela J Bjorkman, Saurabh Mehandru, Paul D Bieniasz, Marina Caskey, Michel C Nussenzweig.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has infected 78 million individuals and is responsible for over 1.7 million deaths to date. Infection is associated with development of variable levels of antibodies with neutralizing activity that can protect against infection in animal models1,2. Antibody levels decrease with time, but the nature and quality of the memory B cells that would be called upon to produce antibodies upon re-infection has not been examined. Here we report on the humoral memory response in a cohort of 87 individuals assessed at 1.3 and 6.2 months after infection. We find that IgM, and IgG anti-SARS-CoV-2 spike protein receptor binding domain (RBD) antibody titres decrease significantly with IgA being less affected. Concurrently, neutralizing activity in plasma decreases by fivefold in pseudotype virus assays. In contrast, the number of RBD-specific memory B cells is unchanged. Memory B cells display clonal turnover after 6.2 months, and the antibodies they express have greater somatic hypermutation, increased potency and resistance to RBD mutations, indicative of continued evolution of the humoral response. Analysis of intestinal biopsies obtained from asymptomatic individuals 4 months after the onset of coronavirus disease-2019 (COVID-19), using immunofluorescence, or polymerase chain reaction, revealed persistence of SARS-CoV-2 nucleic acids and immunoreactivity in the small bowel of 7 out of 14 volunteers. We conclude that the memory B cell response to SARS-CoV-2 evolves between 1.3 and 6.2 months after infection in a manner that is consistent with antigen persistence.

DOI: https://doi.org/10.1038/s41586-021-03207-w
Nat Immunol. 2021 Jan 18.

Distinct metabolic programs established in the thymus control effector functions of γδ T cell subsets in tumor microenvironments.

Noella Lopes, Claire McIntyre, Stefania Martin, Mathilde Raverdeau, Nital Sumaria, Ayano C Kohlgruber, Gina J Fiala, Leandro Z Agudelo, Lydia Dyck, Harry Kane, Aaron Douglas, Stephen Cunningham, Hannah Prendeville, Roisin Loftus, Colleen Carmody, Philippe Pierre, Manolis Kellis, Michael Brenner, Rafael J Argüello, Bruno Silva-Santos, Daniel J Pennington, Lydia Lynch.

Metabolic programming controls immune cell lineages and functions, but little is known about γδ T cell metabolism. Here, we found that γδ T cell subsets making either interferon-γ (IFN-γ) or interleukin (IL)-17 have intrinsically distinct metabolic requirements. Whereas IFN-γ+ γδ T cells were almost exclusively dependent on glycolysis, IL-17+ γδ T cells strongly engaged oxidative metabolism, with increased mitochondrial mass and activity. These distinct metabolic signatures were surprisingly imprinted early during thymic development and were stably maintained in the periphery and within tumors. Moreover, pro-tumoral IL-17+ γδ T cells selectively showed high lipid uptake and intracellular lipid storage and were expanded in obesity and in tumors of obese mice. Conversely, glucose supplementation enhanced the antitumor functions of IFN-γ+ γδ T cells and reduced tumor growth upon adoptive transfer. These findings have important implications for the differentiation of effector γδ T cells and their manipulation in cancer immunotherapy.

DOI: https://doi.org/10.1038/s41590-020-00848-3
Science. 2021 01 22. 371(6527): 400-405

Host succinate is an activation signal for Salmonella virulence during intracellular infection.

Gili Rosenberg, Dror Yehezkel, Dotan Hoffman, Camilla Ciolli Mattioli, Moran Fremder, Hadar Ben-Arosh, Leia Vainman, Noa Nissani, Shelly Hen-Avivi, Shirley Brenner, Maxim Itkin, Sergey Malitsky, Ehud Ohana, Noa Bossel Ben-Moshe, Roi Avraham.

Key to the success of intracellular pathogens is the ability to sense and respond to a changing host cell environment. Macrophages exposed to microbial products undergo metabolic changes that drive inflammatory responses. However, the role of macrophage metabolic reprogramming in bacterial adaptation to the intracellular environment has not been explored. Here, using metabolic profiling and dual RNA sequencing, we show that succinate accumulation in macrophages is sensed by intracellular Salmonella Typhimurium (S. Tm) to promote antimicrobial resistance and type III secretion. S Tm lacking the succinate uptake transporter DcuB displays impaired survival in macrophages and in mice. Thus, S Tm co-opts the metabolic reprogramming of infected macrophages as a signal that induces its own virulence and survival, providing an additional perspective on metabolic host-pathogen cross-talk.

DOI: https://doi.org/10.1126/science.aba8026
ACS Biomater Sci Eng. 2020 Apr 13. 6(4): 2159-2166

Regenerating Antithrombotic Surfaces through Nucleic Acid Displacement.

Stephanie L McNamara, Yevgeny Brudno, Alex B Miller, Hyun Oki Ham, Michael Aizenberg, Elliot L Chaikof, David J Mooney.

  Blood-contacting devices are commonly coated with antithrombotic agents to prevent clot formation and to extend the lifespan of the device. However, in vivo degradation of these bioactive surface agents ultimately limits device efficacy and longevity. Here, a regenerative antithrombotic catheter surface treatment is developed using oligodeoxynucleotide (ODN) toehold exchange. ODN strands modified to carry antithrombotic payloads can inhibit the thrombin enzyme when bound to a surface and exchange with rapid kinetics over multiple cycles, even while carrying large payloads. The surface-bound ODNs inhibit thrombin activity to significantly reduce fibrinogen cleavage and fibrin formation, and this effect is sustained after ODN exchange of the surface-bound strands with a fresh antithrombotic payload. This study presents a unique strategy for achieving a continuous antithrombotic state for blood-contacting devices using an ODN-based regeneration method.

Keywords:  antithrombotic; blood-contacting devices; oligonucleotides; refillable surface coatings; toehold exchange

DOI:  https://doi.org/10.1021/acsbiomaterials.0c00038
Nat Commun. 2021 01 18. 12(1): 411

Ionic shape-morphing microrobotic end-effectors for environmentally adaptive targeting, releasing, and sampling.

Zhiqiang Zheng, Huaping Wang, Lixin Dong, Qing Shi, Jianing Li, Tao Sun, Qiang Huang, Toshio Fukuda.

Shape-morphing uses a single actuation source for complex-task-oriented multiple patterns generation, showing a more promising way than reconfiguration, especially for microrobots, where multiple actuators are typically hardly available. Environmental stimuli can induce additional causes of shape transformation to compensate the insufficient space for actuators and sensors, which enriches the shape-morphing and thereby enhances the function and intelligence as well. Here, making use of the ionic sensitivity of alginate hydrogel microstructures, we present a shape-morphing strategy for microrobotic end-effectors made from them to adapt to different physiochemical environments. Pre-programmed hydrogel crosslinks were embedded in different patterns within the alginate microstructures in an electric field using different electrode configurations. These microstructures were designed for accomplishing tasks such as targeting, releasing and sampling under the control of a magnetic field and environmental ionic stimuli. In addition to structural flexibility and environmental ion sensitivity, these end-effectors are also characterized by their complete biodegradability and versatile actuation modes. The latter includes global locomotion of the whole end-effector by self-trapping magnetic microspheres as a hitch-hiker and the local opening and closing of the jaws using encapsulated nanoparticles based on local ionic density or pH values. The versatility was demonstrated experimentally in both in vitro environments and ex vivo in a gastrointestinal tract. Global locomotion was programmable and the local opening and closing was achieved by changing the ionic density or pH values. This 'structural intelligence' will enable strategies for shape-morphing and functionalization, which have attracted growing interest for applications in minimally invasive medicine, soft robotics, and smart materials.

DOI: https://doi.org/10.1038/s41467-020-20697-w
ACS Nano. 2021 Jan 22.

Design of SARS-CoV-2 hFc-Conjugated Receptor-Binding Domain mRNA Vaccine Delivered via Lipid Nanoparticles.

Uri Elia, Srinivas Ramishetti, Ronit Rosenfeld, Niels Dammes, Erez Bar-Haim, Gonna Somu Naidu, Efi Makdasi, Yfat Yahalom-Ronen, Hadas Tamir, Nir Paran, Ofer Cohen, Dan Peer.

  Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the causal agent of COVID-19 and stands at the center of the current global human pandemic, with death toll exceeding one million. The urgent need for a vaccine has led to the development of various immunization approaches. mRNA vaccines represent a cell-free, simple, and rapid platform for immunization, and therefore have been employed in recent studies toward the development of a SARS-CoV-2 vaccine. Herein, we present the design of an mRNA vaccine, based on lipid nanoparticles (LNPs)-encapsulated SARS-CoV-2 human Fc-conjugated receptor-binding domain (RBD-hFc). Several ionizable lipids have been evaluated in vivo in a luciferase (luc) mRNA reporter assay, and two leading LNPs formulations have been chosen for the subsequent RBD-hFc mRNA vaccine strategy. Intramuscular administration of LNP RBD-hFc mRNA elicited robust humoral response, a high level of neutralizing antibodies and a Th1-biased cellular response in BALB/c mice. The data in the current study demonstrate the potential of these lipids as promising candidates for LNP-based mRNA vaccines in general and for a COVID19 vaccine in particular.

Keywords:  COVID-19; SARS-CoV-2; ionizable lipids; lipid nanoparticles; mRNA vaccine

DOI:  https://doi.org/10.1021/acsnano.0c10180
ACS Nano. 2021 Jan 20.

Immunogenic Hybrid Nanovesicles of Liposomes and Tumor-Derived Nanovesicles for Cancer Immunochemotherapy.

Mei Hu, Jiao Zhang, Li Kong, Yulin Yu, Qian Hu, Ting Yang, Yi Wang, Kun Tu, Qi Qiao, Xianya Qin, Zhiping Zhang.

  Exploring a rational delivery system of integrating chemotherapy with immunotherapy to broaden benefits of cancer immunochemotherapy is still under challenge. Herein, we developed doxorubicin (DOX)-loaded biomimetic hybrid nanovesicles (DOX@LINV) via fusing artificial liposomes (LIPs) with tumor-derived nanovesicles (TNVs) for combinational immunochemotherapy. DOX@LINV with a homologous targeting ability could deliver DOX to tumor tissue and elicit an effective immunogenic cell death response to improve the immunogenicity of a tumor. Meanwhile, the preserved tumor antigens and endogenous danger signals in DOX@LINV activated dendritic cells and induced a subsequent antigen-specific T cell immune response. DOX@LINV displayed a specific antitumor effect on murine melanoma, Lewis lung cancer, and 4T1 breast cancer based on the infiltration of effector immune cells and improvement of the immunosuppressive tumor microenvironment. Furthermore, the combination of DOX@LINV with immune checkpoint inhibitor amplified antitumor efficacy with 33.3% of the mice being tumor-free. Therefore, the hybrid LINV is a promising drug delivery platform with a boosted antitumor immune response for effective immunochemotherapy.

Keywords:  hybrid nanovesicles; immunochemotherapy; immunogenic cell death; liposomes; tumor-derived nanovesicles

DOI:  https://doi.org/10.1021/acsnano.0c09681
Adv Mater. 2021 Jan 22. e2006570

Biomimetic Design of Mitochondria-Targeted Hybrid Nanozymes as Superoxide Scavengers.

Yue Zhang, Anila Khalique, Xinchen Du, Zhanxia Gao, Jin Wu, Xiangyun Zhang, Ran Zhang, Zhiyuan Sun, Qiqi Liu, Zhelong Xu, Adam C Midgley, Lianyong Wang, Xiyun Yan, Jie Zhuang, Deling Kong, Xinglu Huang.

  Development of enzyme mimics for the scavenging of excessive mitochondrial superoxide (O2 •- ) can serve as an effective strategy in the treatment of many diseases. Here, protein reconstruction technology and nanotechnology is taken advantage of to biomimetically create an artificial hybrid nanozyme. These nanozymes consist of ferritin-heavy-chain-based protein as the enzyme scaffold and a metal nanoparticle core as the enzyme active center. This artificial cascade nanozyme possesses superoxide dismutase- and catalase-like activities and also targets mitochondria by overcoming multiple biological barriers. Using cardiac ischemia-reperfusion animal models, the protective advantages of the hybrid nanozymes are demonstrated in vivo during mitochondrial oxidative injury and in the recovery of heart functionality following infarction via systemic delivery and localized release from adhesive hydrogels (i.e., cardiac patch), respectively. This study illustrates a de novo design strategy in the development of enzyme mimics and provides a promising therapeutic option for alleviating oxidative damage in regenerative medicine.

Keywords:  de novo design; mitochondria; nanozymes; protein scaffolds; superoxide scavengers

DOI:  https://doi.org/10.1002/adma.202006570
Nat Cancer. 2020 Oct;1(10): 998-1009

Metabolic adaptation of acute lymphoblastic leukemia to the central nervous system microenvironment is dependent on Stearoyl CoA desaturase.

Angela Maria Savino, Sara Isabel Fernandes, Orianne Olivares, Anna Zemlyansky, Antony Cousins, Elke K Markert, Shani Barel, Ifat Geron, Liron Frishman, Yehudit Birger, Cornelia Eckert, Sergey Tumanov, Gillian MacKay, Jurre J Kamphorst, Pawel Herzyk, Jonatan Fernández-García, Ifat Abramovich, Inbal Mor, Michela Bardini, Ersilia Barin, Sudha Janaki-Raman, Justin R Cross, Michael G Kharas, Eyal Gottlieb, Shai Izraeli, Christina Halsey.

  Metabolic reprogramming is a key hallmark of cancer, but less is known about metabolic plasticity of the same tumor at different sites. Here, we investigated the metabolic adaptation of leukemia in two different microenvironments, the bone marrow and the central nervous system (CNS). We identified a metabolic signature of fatty-acid synthesis in CNS leukemia, highlighting Stearoyl-CoA desaturase (SCD1) as a key player. In vivo SCD1 overexpression increases CNS disease, whilst genetic or pharmacological inhibition of SCD1 decreases CNS load. Overall, we demonstrated that leukemic cells dynamically rewire metabolic pathways to suit local conditions and that targeting these adaptations can be exploited therapeutically.

Keywords:  SCD1; acute lymphoblastic leukemia; central nervous system; fatty acid synthesis; metabolic reprogramming

DOI:  https://doi.org/10.1038/s43018-020-00115-2
ACS Nano. 2021 Jan 19.

Integration of Multitargeted Polymer-Based Contrast Agents with Photoacoustic Computed Tomography: An Imaging Technique to Visualize Breast Cancer Intratumor Heterogeneity.

Lei Li, Deepanjali Patil, Greg Petruncio, Kathleen K Harnden, Jisha V Somasekharan, Mikell Paige, Lihong V Wang, Carolina Salvador-Morales.

  One of the primary challenges in breast cancer diagnosis and treatment is intratumor heterogeneity (ITH), i.e., the coexistence of different genetically and epigenetically distinct malignant cells within the same tumor. Thus, the identification of ITH is critical for designing better treatments and hence to increase patient survival rates. Herein, we report a noninvasive hybrid imaging technology that integrates multitargeted and multiplexed patchy polymeric photoacoustic contrast agents (MTMPPPCAs) with single-impulse panoramic photoacoustic computed tomography (SIP-PACT). The target specificity ability of MTMPPPCAs to distinguish estrogen and progesterone receptor-positive breast tumors was demonstrated through both fluorescence and photoacoustic measurements and validated by tissue pathology analysis. This work provides the proof-of-concept of the MTMPPPCAs/SIP-PACT system to identify ITH in nonmetastatic tumors, with both high molecular specificity and real-time detection capability.

Keywords:  breast cancer; intratumor heterogeneity; multitargeted; patchy particles; single-impulse panoramic photoacoustic computed tomography

DOI:  https://doi.org/10.1021/acsnano.0c05893
Science. 2021 01 22. pii: eaba6500. [Epub ahead of print]371(6527):

Developmental cell programs are co-opted in inflammatory skin disease.

Gary Reynolds, Peter Vegh, James Fletcher, Elizabeth F M Poyner, Emily Stephenson, Issac Goh, Rachel A Botting, Ni Huang, Bayanne Olabi, Anna Dubois, David Dixon, Kile Green, Daniel Maunder, Justin Engelbert, Mirjana Efremova, Krzysztof Polański, Laura Jardine, Claire Jones, Thomas Ness, Dave Horsfall, Jim McGrath, Christopher Carey, Dorin-Mirel Popescu, Simone Webb, Xiao-Nong Wang, Ben Sayer, Jong-Eun Park, Victor A Negri, Daria Belokhvostova, Magnus D Lynch, David McDonald, Andrew Filby, Tzachi Hagai, Kerstin B Meyer, Akhtar Husain, Jonathan Coxhead, Roser Vento-Tormo, Sam Behjati, Steven Lisgo, Alexandra-Chloé Villani, Jaume Bacardit, Philip H Jones, Edel A O'Toole, Graham S Ogg, Neil Rajan, Nick J Reynolds, Sarah A Teichmann, Fiona M Watt, Muzlifah Haniffa.

The skin confers biophysical and immunological protection through a complex cellular network established early in embryonic development. We profiled the transcriptomes of more than 500,000 single cells from developing human fetal skin, healthy adult skin, and adult skin with atopic dermatitis and psoriasis. We leveraged these datasets to compare cell states across development, homeostasis, and disease. Our analysis revealed an enrichment of innate immune cells in skin during the first trimester and clonal expansion of disease-associated lymphocytes in atopic dermatitis and psoriasis. We uncovered and validated in situ a reemergence of prenatal vascular endothelial cell and macrophage cellular programs in atopic dermatitis and psoriasis lesional skin. These data illustrate the dynamism of cutaneous immunity and provide opportunities for targeting pathological developmental programs in inflammatory skin diseases.

DOI: https://doi.org/10.1126/science.aba6500
Nat Commun. 2021 01 18. 12(1): 409

Low-dose decitabine priming endows CAR T cells with enhanced and persistent antitumour potential via epigenetic reprogramming.

Yao Wang, Chuan Tong, Hanren Dai, Zhiqiang Wu, Xiao Han, Yelei Guo, Deyun Chen, Jianshu Wei, Dongdong Ti, Zongzhi Liu, Qian Mei, Xiang Li, Liang Dong, Jing Nie, Yajing Zhang, Weidong Han.

Insufficient eradication capacity and dysfunction are common occurrences in T cells that characterize cancer immunotherapy failure. De novo DNA methylation promotes T cell exhaustion, whereas methylation inhibition enhances T cell rejuvenation in vivo. Decitabine, a DNA methyltransferase inhibitor approved for clinical use, may provide a means of modifying exhaustion-associated DNA methylation programmes. Herein, anti-tumour activities, cytokine production, and proliferation are enhanced in decitabine-treated chimeric antigen receptor T (dCAR T) cells both in vitro and in vivo. Additionally, dCAR T cells can eradicate bulky tumours at a low-dose and establish effective recall responses upon tumour rechallenge. Antigen-expressing tumour cells trigger higher expression levels of memory-, proliferation- and cytokine production-associated genes in dCAR T cells. Tumour-infiltrating dCAR T cells retain a relatively high expression of memory-related genes and low expression of exhaustion-related genes in vivo. In vitro administration of decitabine may represent an option for the generation of CAR T cells with improved anti-tumour properties.

DOI: https://doi.org/10.1038/s41467-020-20696-x
Adv Healthc Mater. 2021 Jan 18. e2001718

Recent Advances of DNA Nanostructure-Based Cell Membrane Engineering.

Lingyu Feng, Jiang Li, Jielin Sun, Lihua Wang, Chunhai Fan, Jianlei Shen.

  Materials that can regulate the composition and structure of the cell membrane to fabricate engineered cells with defined functions are in high demand. Compared with other biomolecules, DNA has unique advantages in cell membrane engineering due to its excellent programmability and biocompatibility. Especially, the near-atomic scale precision of DNA nanostructures facilitates the investigation of structure-property relations on the cell membrane. In this review, first the state of the art of functional DNA nanostructures is summarized, and then the overview of the use of DNA nanostructures to engineer the cell membrane is presented. Subsequently, applications of DNA nanostructures in modifying cell membrane morphology, controlling ions transport, and synthesizing high precise liposomes are highlighted. Finally, the challenges and outlook on using DNA nanostructures for cell membrane engineering are discussed.

Keywords:  DNA nanostructures; DNA origami; artificial nanopores; cell membranes; liposomes

DOI:  https://doi.org/10.1002/adhm.202001718
Nat Commun. 2021 01 18. 12(1): 420

Core transcription regulatory circuitry orchestrates corneal epithelial homeostasis.

Mingsen Li, Huaxing Huang, Lingyu Li, Chenxi He, Liqiong Zhu, Huizhen Guo, Li Wang, Jiafeng Liu, Siqi Wu, Jingxin Liu, Tao Xu, Zhen Mao, Nan Cao, Kang Zhang, Fei Lan, Junjun Ding, Jin Yuan, Yizhi Liu, Hong Ouyang.

Adult stem cell identity, plasticity, and homeostasis are precisely orchestrated by lineage-restricted epigenetic and transcriptional regulatory networks. Here, by integrating super-enhancer and chromatin accessibility landscapes, we delineate core transcription regulatory circuitries (CRCs) of limbal stem/progenitor cells (LSCs) and find that RUNX1 and SMAD3 are required for maintenance of corneal epithelial identity and homeostasis. RUNX1 or SMAD3 depletion inhibits PAX6 and induces LSCs to differentiate into epidermal-like epithelial cells. RUNX1, PAX6, and SMAD3 (RPS) interact with each other and synergistically establish a CRC to govern the lineage-specific cis-regulatory atlas. Moreover, RUNX1 shapes LSC chromatin architecture via modulating H3K27ac deposition. Disturbance of RPS cooperation results in cell identity switching and dysfunction of the corneal epithelium, which is strongly linked to various human corneal diseases. Our work highlights CRC TF cooperativity for establishment of stem cell identity and lineage commitment, and provides comprehensive regulatory principles for human stratified epithelial homeostasis and pathogenesis.

DOI: https://doi.org/10.1038/s41467-020-20713-z
Nano Lett. 2021 Jan 21.

A Biomimetic Drug Delivery System by Integrating Grapefruit Extracellular Vesicles and Doxorubicin-Loaded Heparin-Based Nanoparticles for Glioma Therapy.

Wenbo Niu, Qian Xiao, Xuejiao Wang, Junqiao Zhu, Jinheng Li, Xiaomei Liang, Yingming Peng, Chentian Wu, Ruojing Lu, Yao Pan, Jiamao Luo, Xinxian Zhong, Haoqi He, Zhili Rong, Jun-Bing Fan, Ying Wang.

  Existing nanoparticle-mediated drug delivery systems for glioma systemic chemotherapy remain a great challenge due to poor delivery efficiency resulting from the blood brain barrier/blood-(brain tumor) barrier (BBB/BBTB) and insufficient tumor penetration. Here, we demonstrate a distinct design by patching doxorubicin-loaded heparin-based nanoparticles (DNs) onto the surface of natural grapefruit extracellular vesicles (EVs), to fabricate biomimetic EV-DNs, achieving efficient drug delivery and thus significantly enhancing antiglioma efficacy. The patching strategy allows the unprecedented 4-fold drug loading capacity compared to traditional encapsulation for EVs. The biomimetic EV-DNs are enabled to bypass BBB/BBTB and penetrate into glioma tissues by receptor-mediated transcytosis and membrane fusion, greatly promoting cellular internalization and antiproliferation ability as well as extending circulation time. We demonstrate that a high-abundance accumulation of EV-DNs can be detected at glioma tissues, enabling the maximal brain tumor uptake of EV-DNs and great antiglioma efficacy in vivo.

Keywords:  biomimetic delivery system; glioma therapy; grapefruit extracellular vesicles; nanoparticles

DOI:  https://doi.org/10.1021/acs.nanolett.0c04753
ACS Nano. 2021 Jan 19.

Rapid Development of SARS-CoV-2 Spike Protein Receptor-Binding Domain Self-Assembled Nanoparticle Vaccine Candidates.

Yin-Feng Kang, Cong Sun, Zhen Zhuang, Run-Yu Yuan, Qingbing Zheng, Jiang-Ping Li, Ping-Ping Zhou, Xin-Chun Chen, Zhe Liu, Xiao Zhang, Xiao-Hui Yu, Xiang-Wei Kong, Qian-Ying Zhu, Qian Zhong, Miao Xu, Nan-Shan Zhong, Yi-Xin Zeng, Guo-Kai Feng, Changwen Ke, Jin-Cun Zhao, Mu-Sheng Zeng.

  The coronavirus disease pandemic of 2019 (COVID-19) caused by the novel SARS-CoV-2 coronavirus resulted in economic losses and threatened human health worldwide. The pandemic highlights an urgent need for a stable, easily produced, and effective vaccine. SARS-CoV-2 uses the spike protein receptor-binding domain (RBD) to bind its cognate receptor, angiotensin-converting enzyme 2 (ACE2), and initiate membrane fusion. Thus, the RBD is an ideal target for vaccine development. In this study, we designed three different RBD-conjugated nanoparticle vaccine candidates, namely, RBD-Ferritin (24-mer), RBD-mi3 (60-mer), and RBD-I53-50 (120-mer), via covalent conjugation using the SpyTag-SpyCatcher system. When mice were immunized with the RBD-conjugated nanoparticles (NPs) in conjunction with the AddaVax or Sigma Adjuvant System, the resulting antisera exhibited 8- to 120-fold greater neutralizing activity against both a pseudovirus and the authentic virus than those of mice immunized with monomeric RBD. Most importantly, sera from mice immunized with RBD-conjugated NPs more efficiently blocked the binding of RBD to ACE2 in vitro, further corroborating the promising immunization effect. Additionally, the vaccine has distinct advantages in terms of a relatively simple scale-up and flexible assembly. These results illustrate that the SARS-CoV-2 RBD-conjugated nanoparticles developed in this study are a competitive vaccine candidate and that the carrier nanoparticles could be adopted as a universal platform for a future vaccine development.

Keywords:  SARS-CoV-2; SpyTag-SpyCatcher; covalent conjugation; nanoparticles; receptor binding domain; vaccine

DOI:  https://doi.org/10.1021/acsnano.0c08379
Nat Commun. 2021 Jan 22. 12(1): 555

CRISPR technologies and the search for the PAM-free nuclease.

Daphne Collias, Chase L Beisel.

The ever-expanding set of CRISPR technologies and their programmable RNA-guided nucleases exhibit remarkable flexibility in DNA targeting. However, this flexibility comes with an ever-present constraint: the requirement for a protospacer adjacent motif (PAM) flanking each target. While PAMs play an essential role in self/nonself discrimination by CRISPR-Cas immune systems, this constraint has launched a far-reaching expedition for nucleases with relaxed PAM requirements. Here, we review ongoing efforts toward realizing PAM-free nucleases through natural ortholog mining and protein engineering. We also address potential consequences of fully eliminating PAM recognition and instead propose an alternative nuclease repertoire covering all possible PAM sequences.

DOI: https://doi.org/10.1038/s41467-020-20633-y
Mol Ther Nucleic Acids. 2021 Mar 05. 23 406-417

Adeno-associated virus-mediated delivery of anti-miR-199a tough decoys attenuates cardiac hypertrophy by targeting PGC-1alpha.

Hualin Yan, Hong Wang, Xiaoxia Zhu, Jianbo Huang, Yifei Li, Kaiyu Zhou, Yimin Hua, Feng Yan, Da-Zhi Wang, Yan Luo.

  MicroRNAs (miRNAs) are important regulators in the process of cardiac hypertrophy and heart failure. Previous studies have shown that miR-199a is upregulated in pressure-overload cardiac hypertrophy and that inhibition of miR-199a attenuates cardiac hypertrophy in vitro. However, the therapeutic role of anti-miR-199a treatment in the cardiac hypertrophy in vivo model is less known. Here, we show an efficient and useful method to treat mouse cardiac hypertrophy and restore cardiac function through injection of adeno-associated virus (AAV)-mediated anti-miR-199a tough decoys (TuDs). RNA-seq transcriptome analysis indicated that genes related to cytoplasmic translation and mitochondrial respiratory chain complex assembly were upregulated in anti-miR-199a-treated recovered hearts. We further validated that PGC-1α is the direct target of miR-199a involved in the therapeutic effect and the regulation of the PGC-1α/ERRα axis and that the downstream pathway of mitochondrial fatty acid oxidation and oxidative phosphorylation constitute the underlying mechanism of the restored mitochondrial structure and function in our anti-miR-199a-treated mice. Our study highlights the important regulatory role of miR-199a in cardiac hypertrophy and the value of the AAV-mediated miRNA delivery system.

Keywords:  PGC1-alpha; adeno-associated virus; cardiac hypertrophy; heart failure; microRNA-199a

DOI:  https://doi.org/10.1016/j.omtn.2020.11.007
Nat Commun. 2021 01 19. 12(1): 440

Boosting anti-PD-1 therapy with metformin-loaded macrophage-derived microparticles.

Zhaohan Wei, Xiaoqiong Zhang, Tuying Yong, Nana Bie, Guiting Zhan, Xin Li, Qingle Liang, Jianye Li, Jingjing Yu, Gang Huang, Yuchen Yan, Zelong Zhang, Bixiang Zhang, Lu Gan, Bo Huang, Xiangliang Yang.

The main challenges for programmed cell death 1(PD-1)/PD-1 ligand (PD-L1) checkpoint blockade lie in a lack of sufficient T cell infiltration, tumor immunosuppressive microenvironment, and the inadequate tumor accumulation and penetration of anti-PD-1/PD-L1 antibody. Resetting tumor-associated macrophages (TAMs) is a promising strategy to enhance T-cell antitumor immunity and ameliorate tumor immunosuppression. Here, mannose-modified macrophage-derived microparticles (Man-MPs) loading metformin (Met@Man-MPs) are developed to efficiently target to M2-like TAMs to repolarize into M1-like phenotype. Met@Man-MPs-reset TAMs remodel the tumor immune microenvironment by increasing the recruitment of CD8+ T cells into tumor tissues and decreasing immunosuppressive infiltration of myeloid-derived suppressor cells and regulatory T cells. More importantly, the collagen-degrading capacity of Man-MPs contributes to the infiltration of CD8+ T cells into tumor interiors and enhances tumor accumulation and penetration of anti-PD-1 antibody. These unique features of Met@Man-MPs contribute to boost anti-PD-1 antibody therapy, improving anticancer efficacy and long-term memory immunity after combination treatment. Our results support Met@Man-MPs as a potential drug to improve tumor resistance to anti-PD-1 therapy.

DOI: https://doi.org/10.1038/s41467-020-20723-x
Nano Lett. 2021 Jan 16.

Oral Administration of Starting Materials for In Vivo Synthesis of Antibacterial Gold Nanoparticles for Curing Remote Infections.

Le Wang, Junchuan Yang, Sixiang Li, Qizhen Li, Shaoqin Liu, Wenfu Zheng, Xingyu Jiang.

  Oral administration is a facile and safe way for medication. However, most of the reported nanomedicines could not be taken orally, partially due to their unsatisfied stability, poor absorbance, or toxicity in the gastrointestinal tract. Here, we demonstrate that we could robustly synthesize gold nanoparticles (GNPs) in vivo by orally administering two starting materials, tetrachloroauric acid and aminophenyl boronic acid (ABA). The ABA-activated GNPs (A-GNPs) synthesized in vivo could be absorbed by the gastrointestinal tract and reach the remote infection lesions such as peritonitis caused by multidrug resistant (MDR) bacteria in mice. The A-GNPs exhibit excellent antibacterial efficacy (MIC, 3 μg/mL), long half-life (16-17 h), effective clearance (residual concentration is near 0 within 72 h), and high biosafety (safe dose/effective dose, 8 times). Our study is a pioneering attempt for synthesizing and taking nanomedicines orally just like preparing and drinking a cocktail.

Keywords:  Antibacterial; Biosafety; Gold nanoparticles; In vivo synthesis; Oral administration

DOI:  https://doi.org/10.1021/acs.nanolett.0c04578
Acta Biomater. 2021 Jan 16. pii: S1742-7061(21)00040-4. [Epub ahead of print]

Hydrogel microspheres for spatiotemporally controlled delivery of RNA and silencing gene expression within scaffold-free tissue engineered constructs: Spatiotemporally controlled gene expression within MSC aggregates.

Alexandra McMillan, Minh Khanh Nguyen, Cong Truc Huynh, Samantha M Sarett, Peilin Ge, Melanie Chetverikova, Kien Nguyen, David Grosh, Craig L Duvall, Eben Alsberg.

  Delivery systems for controlled release of RNA interference (RNAi) molecules, including small interfering (siRNA) and microRNA (miRNA), have the potential to direct stem cell differentiation for regenerative musculoskeletal applications. To date, localized RNA delivery platforms in this area have focused predominantly on bulk scaffold-based approaches, which can interfere with cell-cell interactions important for recapitulating some native musculoskeletal developmental and healing processes in tissue regeneration strategies. In contrast, scaffold-free, high density human mesenchymal stem cell (hMSC) aggregates may provide an avenue for creating a more biomimetic microenvironment. Here, photocrosslinkable dextran microspheres (MS) encapsulating siRNA-micelles were prepared via an aqueous emulsion method and incorporated within hMSC aggregates for localized and sustained delivery of bioactive siRNA. siRNA-micelles released from MS in a sustained fashion over the course of 28 days, and the released siRNA retained its ability to transfect cells for gene silencing. Incorporation of fluorescently labeled siRNA (siGLO)-laden MS within hMSC aggregates exhibited tunable siGLO delivery and uptake by stem cells. Incorporation of MS loaded with siRNA targeting green fluorescent protein (siGFP) within GFP-hMSC aggregates provided sustained presentation of siGFP within the constructs and prolonged GFP silencing for up to 15 days. This platform system enables sustained gene silencing within stem cell aggregates and thus shows great potential in tissue regeneration applications.

Keywords:  Mesenchymal stem cells; RNA interference; aggregates; gene therapy; sustained delivery

DOI:  https://doi.org/10.1016/j.actbio.2021.01.013
ACS Biomater Sci Eng. 2019 Oct 14. 5(10): 4920-4936

How Functionalized Surfaces Can Inhibit Bacterial Adhesion and Viability.

Fiorela Ghilini, Diego E Pissinis, Alejandro Miñán, Patricia L Schilardi, Carolina Diaz.

  Device-associated infections (DAI) remain a serious concern in modern healthcare. Bacterial attachment to a surface is the first step in biofilm formation, which is one of the main causes of DAIs. The development of materials capable of preventing or inhibiting bacterial attachment constitutes a promising approach to deal with this problem. The multifactorial nature of biofilm maturation and antibiotic resistance directs the research for multitargeted or combinatorial therapeutic approaches. One attractive strategy is the modification or the engineering of surfaces in order to provide antiadhesive and/or antimicrobial properties. Currently, several different approaches that involve physical and chemical surface modification deliver some possible alternatives to achieve this goal. The engineered surfaces can be coated with molecules capable of inhibiting the bacterial adhesion or with active agents that kill microorganisms. In addition, surfaces can also be modified in order to be stimuli-responsive, responding to a particular trigger and then delivering the consequent antimicrobial outcome. Here, we review the prevailing strategies to modify surfaces in order to create an antimicrobial surface and discuss how different surface functionalization can affect bacterial adhesion and/or viability.

Keywords:  antibacterial surfaces; bacterial adhesion; bacterial viability; biofilm; indwelling devices; surface modification

DOI:  https://doi.org/10.1021/acsbiomaterials.9b00849