Neutrophil Extracellular Traps and Its Implications in Inflammation: An Overview

Frontiers in Immunology, Journal Year: 2017, Volume and Issue: 8

Published: Feb. 6, 2017

In addition to physical barriers, neutrophils are considered a part of the first line immune defense. They can be found in bloodstream, with lifespan 6-8 hours, and tissue, where they last up seven days. The mechanisms that utilize for host defense phagocytosis, degranulation, cytokine production and, most recently described, neutrophil extracellular trap (NET) production. NETs DNA structures released due chromatin decondensation spreading, thus occupy 3-5 times volume condensed chromatin. Several proteins adhere NETs, including histones over 30 components primary secondary granules, among them bactericidal activity such as elastase, myeloperoxidase, cathepsin G, lactoferrin, pentraxin 3, gelatinase, proteinase LL-37, peptidoglycan-binding others able destroy virulence factors. Three models NETosis known date. a) Suicidal NETosis, duration 2-4 is best described model. b) vital nuclear release, release without exhibiting loss or plasma membrane within 5-60 minutes, it independent ROS Raf/MERK/ERK pathway. c). final type Vital mitochondrial dependent on produced after stimuli GM-CSF LPS. Recent research has revealed more sophisticated cells precisely regulate their granular enzymes by ion fluxes immunomodulatory cytokines chemokines interact various system. Therefore, play key role autoimmunity autoinflammatory metabolic diseases. this review, we intend show two roles played neutrophils: against microorganisms contributor pathogenesis illnesses, autoimmune,

Language: Английский

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Delivery strategies of the CRISPR-Cas9 gene-editing system for therapeutic applications

Journal of Controlled Release, Journal Year: 2017, Volume and Issue: 266, P. 17 - 26

Published: Sept. 11, 2017

Language: Английский

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CRISPR/Cas9-Based Genome Editing for Disease Modeling and Therapy: Challenges and Opportunities for Nonviral Delivery

Chemical Reviews, Journal Year: 2017, Volume and Issue: 117(15), P. 9874 - 9906

Published: June 22, 2017

Genome editing offers promising solutions to genetic disorders by DNA sequences or modulating gene expression. The clustered regularly interspaced short palindromic repeats (CRISPR)/associated protein 9 (CRISPR/Cas9) technology can be used edit single multiple genes in a wide variety of cell types and organisms vitro vivo. Herein, we review the rapidly developing CRISPR/Cas9-based technologies for disease modeling correction recent progress toward Cas9/guide RNA (gRNA) delivery based on viral nonviral vectors. We discuss relative merits delivering genome elements form DNA, mRNA, protein, opportunities combining transgene encoding Cas9 with gRNA. highlight lessons learned from past three decades consider their applicability CRISPR/Cas9 delivery. also include discussion bioinformatics tools gRNA design chemical modifications Finally, extracellular intracellular barriers propose strategies that may overcome these realize clinical potential editing.

Language: Английский

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Naturally Occurring Off-Switches for CRISPR-Cas9

Cell, Journal Year: 2016, Volume and Issue: 167(7), P. 1829 - 1838.e9

Published: Dec. 1, 2016

Language: Английский

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CRISPR/Cas9 editing of endogenous banana streak virus in the B genome of Musa spp. overcomes a major challenge in banana breeding

Communications Biology, Journal Year: 2019, Volume and Issue: 2(1)

Published: Jan. 31, 2019

Abstract Presence of the integrated endogenous banana streak virus (eBSV) in B genome plantain (AAB) is a major challenge for breeding and dissemination hybrids. As eBSV activates into infectious viral particles under stress, progenitor Musa balbisiana its derivants, having at least one genome, cannot be used as parents crop improvement. Here, we report strategy to inactivate by editing sequences. The regenerated genome-edited events Gonja Manjaya showed mutations targeted sites with potential prevent proper transcription or/and translational functional proteins. Seventy-five percent edited remained asymptomatic comparison non-edited control plants water stress conditions, confirming inactivation particles. This study paves way improvement germplasm use programs produce hybrids that can globally disseminated.

Language: Английский

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In Vivo Excision of HIV-1 Provirus by saCas9 and Multiplex Single-Guide RNAs in Animal Models

Molecular Therapy, Journal Year: 2017, Volume and Issue: 25(5), P. 1168 - 1186

Published: March 31, 2017

CRISPR-associated protein 9 (Cas9)-mediated genome editing provides a promising cure for HIV-1/AIDS; however, gene delivery efficiency in vivo remains an obstacle to overcome. Here, we demonstrate the feasibility and of excising HIV-1 provirus three different animal models using all-in-one adeno-associated virus (AAV) vector deliver multiplex single-guide RNAs (sgRNAs) plus Staphylococcus aureus Cas9 (saCas9). The quadruplex sgRNAs/saCas9 outperformed duplex integrated cultured neural stem/progenitor cells from Tg26 transgenic mice. Intravenously injected AAV-DJ/8 excised proviral DNA significantly reduced viral RNA expression several organs/tissues In EcoHIV acutely infected mice, intravenously systemic infection, as determined by live bioluminescence imaging. Additionally, this induced efficient excision, PCR genotyping liver, lungs, brain, spleen. Finally, humanized bone marrow/liver/thymus (BLT) mice with chronic successful excision was detected spleen, heart, colon, brain after single intravenous injection AAV-DJ/8. conclusion, solid tissues/organs can be achieved via AAV delivery, significant step toward human clinical trials.Graphical abstract

Language: Английский

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Sequential LASER ART and CRISPR Treatments Eliminate HIV-1 in a Subset of Infected Humanized Mice

Nature Communications, Journal Year: 2019, Volume and Issue: 10(1)

Published: July 2, 2019

Abstract Elimination of HIV-1 requires clearance and removal integrated proviral DNA from infected cells tissues. Here, sequential long-acting slow-effective release antiviral therapy (LASER ART) CRISPR-Cas9 demonstrate viral in latent infectious reservoirs humanized mice. subgenomic fragments, spanning the long terminal repeats Gag gene, are excised vivo, resulting elimination DNA; virus is not detected blood, lymphoid tissue, bone marrow brain by nested digital-droplet PCR as well RNAscope tests. No mediated off-target effects detected. Adoptive transfer human immunocytes dual treated, virus-free animals to uninfected mice fails produce progeny virus. In contrast, readily following sole LASER ART or treatment. These data provide proof-of-concept that permanent possible.

Language: Английский

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CRISPR/Cas9-Mediated Genome Editing of Herpesviruses Limits Productive and Latent Infections

PLoS Pathogens, Journal Year: 2016, Volume and Issue: 12(6), P. e1005701 - e1005701

Published: June 30, 2016

Herpesviruses infect the majority of human population and can cause significant morbidity mortality. Herpes simplex virus (HSV) type 1 causes cold sores herpes keratitis, whereas HSV-2 is responsible for genital herpes. Human cytomegalovirus (HCMV) most common viral congenital defects serious disease in immuno-compromised individuals. Epstein-Barr (EBV) associated with infectious mononucleosis a broad range malignancies, including Burkitt’s lymphoma, nasopharyngeal carcinoma, Hodgkin’s disease, post-transplant lymphomas. persist their host life by establishing latent infection that interrupted periodic reactivation events during which replication occurs. Current antiviral drug treatments target clinical manifestations this productive stage, but they are ineffective at eliminating these viruses from infected host. Here, we set out to combat both herpesvirus infections exploiting CRISPR/Cas9 system genetic elements important fitness. We show effective abrogation HCMV HSV-1 targeting gRNAs essential genes. Simultaneous multiple completely abolished production particles cells. Using same approach, EBV be almost cleared latently EBV-transformed tumor Our studies indicate effectively targeted genomes as potent prophylactic therapeutic anti-viral strategy may used impair clear infection.

Language: Английский

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Magnetically guided non-invasive CRISPR-Cas9/gRNA delivery across blood-brain barrier to eradicate latent HIV-1 infection

Scientific Reports, Journal Year: 2019, Volume and Issue: 9(1)

Published: March 8, 2019

Abstract CRISPR-Cas9/gRNA exhibits therapeutic efficacy against latent human immunodeficiency virus (HIV) genome but the delivery of this cargo to brain remains as a challenge. In research, for first time, we demonstrated magnetically guided non-invasive nano-formulation (NF), composed Cas9/gRNA bound with magneto-electric nanoparticles (MENPs), across blood-brain barrier (BBB) inhibit HIV-1 infection in microglial (hμglia)/HIV (HC69) cells. An optimized ac-magnetic field 60 Oe was applied on NF release from MENPs surface and facilitate cell uptake resulting intracellular inhibition HIV. The outcomes suggested that developed reduced HIV-LTR expression significantly comparison unbound HIV hμglia/HIV These findings were also validated qualitatively using fluorescence microscopy assess microglia We believe CNS (CRISPR/Cas9-gRNA-MENPs) BBB certainly will have clinical utility future personalized nanomedicine manage neuroHIV/AIDS.

Language: Английский

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Monocytes and Macrophages as Viral Targets and Reservoirs

International Journal of Molecular Sciences, Journal Year: 2018, Volume and Issue: 19(9), P. 2821 - 2821

Published: Sept. 18, 2018

Viruses manipulate cell biology to utilize monocytes/macrophages as vessels for dissemination, long-term persistence within tissues and virus replication. enter cells through endocytosis, phagocytosis, macropinocytosis or membrane fusion. These processes play important roles in the mechanisms contributing pathogenesis of these agents establishing viral genome latency. Upon infection, monocytes respond with an elevated expression proinflammatory signalling molecules antiviral responses, is shown case influenza, Chikungunya, human herpes Zika viruses. Human immunodeficiency initiates acute inflammation on site during early stages infection but there a shift M1 M2 at later infection. Cytomegalovirus creates balance between pro- anti-inflammatory by inducing specific phenotype M1/M2 continuum. Despite facilitating inflammation, infected macrophages generally display abolished apoptosis restricted cytopathic effect, which sustains production. The majority viruses discussed this review employ repository certain use productive This focuses adaptations monocytes/macrophages, immune escape, reprogramming response host cells.

Language: Английский

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