Intra-arterial administration of cell-based biological agents for ischemic stroke therapy

Introduction: Ischemic stroke is becoming a primary cause of disability and death worldwide. To date, therapeutic options remain limited focusing on mechanical thrombolysis or administration of thrombolytic agents. However, these therapies do not promote neuroprotection and neuro-restoration of the ischemic area of the brain.

Areas covered: This review highlights the option of minimal invasive, intra-arterial, administration of biological agents for stroke therapy. The authors provide an update of all available studies, discuss issues that influence outcomes and describe future perspectives which aim to improve clinical outcomes. New therapeutic options based on cellular and molecular interactions following an ischemic brain event, will be highlighted.

Expert opinion: Intra-arterial administration of biological agents during trans-catheter thrombolysis or thrombectomy could limit neuronal cell death and facilitate regeneration or neurogenesis following ischemic brain injury. Despite the initial progress, further meticulous studies are needed in order to establish the clinical use of stem cell-induced neuroprotection and neuroregeneration.     

Molecular xenomonitoring and host identification of Leishmania sand fly vectors in a Mediterranean periurban wildlife park

The epidemiological cycle of zoonotic phlebotomine‐borne Leishmania infantum is a complex system in which domestic animals and wildlife interact and participate in its maintenance and transmission. In this study, we combined entomological surveillance, xenomonitoring of L. infantum and identification of host feeding sources of engorged females to investigate the potential contribution of a periurban wildlife park to leishmaniosis in neighbouring residential areas. Overall, 7,309 sand flies were collected in 111 trap‐days during the summers of 2016–2018 in an endemic area in south‐east Spain. Five different sand fly species were captured, with Phlebotomus perniciosus, the main L. infantum vector in this region, representing the most common species. Sand fly distribution was spatially heterogeneous in terms of species, sexes and female physiological stage (unfed, gravid and engorged females) and related to host distribution and management, and environmental features. None of the 602 sand flies analysed for L. infantum infection by kinetoplast real‐time PCR were positive. We used molecular tools to identify the vertebrate hosts of sand flies and identified 17 host species, mainly mammals. Human DNA was not identified in engorged sand flies. This study provides evidence that wildlife parks in south‐east Spain are ideal grounds for sand fly vectors but do not necessarily increase L. infantum infection risk to humans and dogs living in surrounding residential areas. This is probably because vectors feed mostly on non‐L. infantum competent hosts and this should be investigated for a better understanding of the contribution of wildlife parks to the local epidemiology of L. infantum.     

慢病毒沉默PRDX4抑制肺癌细胞A549迁移和侵袭及其机制的探讨北大核心

目的:研究慢病毒沉默过氧化物还原酶4(peroxiredoxin 4,PRDX4)对肺癌细胞迁移和侵袭能力的影响,并探讨其分子机制。方法:采用shRNA干扰技术敲低肺癌细胞系A549中PRDX4的表达,Western blot和qRT-PCR验证转染效果;划痕愈合实验检测细胞迁移能力;Transwell细胞侵袭实验检测细胞侵袭能力;Western blot分析p-ERK1/2、N-cadherin、Vimentin蛋白表达。结果:成功构建沉默PRDX4的肺癌细胞系;慢病毒感染后A549细胞PRDX4蛋白和mRNA表达水平显著低于对照组(P<0.05);与BC组和NC组相比,Sh组细胞迁移、侵袭能力显著降低(P<0.05);与BC组和NC组相比,Sh组细胞p-ERK1/2、N-cadherin、Vimentin蛋白表达水平显著降低。结论:沉默PRDX4可能通过ERK信号通路抑制肺癌细胞的迁移和侵袭。     

Full-length dystrophin restoration via targeted exon integration by AAV-CRISPR in a humanized mouse model of Duchenne muscular dystrophy

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Progress on ocular siRNA gene‐silencing therapy and drug delivery systems

Age‐related macular degeneration (AMD) and glaucoma are global ocular diseases with high blindness rate. RNA interference (RNAi) is being increasingly used in the treatment of these disorders with siRNA drugs, bevasiranib, AGN211745 and PF‐04523655 for AMD, and SYL040012 and QPI‐1007 for glaucoma. Administration routes and vectors of gene drugs affect their therapeutic effect. Compared with the non‐viral vectors, viral vectors have limited payload capacity and potential immunogenicity. This review summarizes the progress of the ocular siRNA gene‐silencing therapy by focusing on siRNA drugs for AMD and glaucoma already used in clinical research, the main routes of drug delivery and the non‐viral vectors for siRNA drugs.     

Lentiviral Vectors and Adeno‐Associated Virus Vectors: Useful Tools for Gene Transfer in Pain Research

Pain, especially chronic pain, has always been a heated point in both basic and clinical researches since it puts heavy burdens on both individuals and the whole society. A better understanding of the role of biological molecules and various ionic channels involved in pain can shed light on the mechanism under pain and advocate the development of pain management. Using viral vectors to transfer specific genes at targeted sites is a promising method for both research and clinical applications. Lentiviral vectors and adeno‐associated virus (AAV) vectors which allow stable and long‐term expression of transgene in non‐dividing cells are widely applied in pain research. In this review, we thoroughly outline the structure, category, advantages and disadvantages and the delivery methods of lentiviral and AAV vectors. The methods through which lentiviral and AAV vectors are delivered to targeted sites are closely related with the sites, level and period of transgene expression. Focus is placed on the various delivery methods applied to deliver vectors to spinal cord and dorsal root ganglion both of which play important roles in primary nociception. Our goal is to provide insight into the features of these two viral vectors and which administration approach can be chosen for different pain researches. Anat Rec, 301:825–836, 2018. © 2017 The Authors. The Anatomical Record published by Wiley Periodicals, Inc. on behalf of American Association of Anatomists.     

慢病毒载体介导siRNA对裸鼠移植人肺腺癌A549抑瘤作用的实验研究

目的研究慢病毒载体介导siRNA抑制survivin基因表达后对人肺腺癌A549细胞体内增殖能力的影响以及对裸鼠移植人肺腺癌的抑瘤活性。方法应用裸鼠成瘤实验测定致瘤能力的改变，采用RT-PCR和Western-blot测定干扰后survivin基因及其蛋白的表达水平；流式细胞术检测肺癌细胞凋亡指数和增殖指数。结果靶向survivin基因的siRNA可以有效抑制survivin基因的表达，使survivin-mRNA表达减少50％-80％，蛋白表达减少60％；流式细胞术检测细胞凋亡率明显提高（P〈0．01），降低了在裸鼠体内成瘤的能力。结论RNA干扰survivin基因表达后明显抑制了人肺腺癌A549细胞的体内增殖能力；siRNA介导的survivin基因下调可明显抑制肿瘤细胞在体内的生长。     

Systemic gene transfer of binding immunoglobulin protein (BiP) prevents disease progression in murine collagen-induced arthritis

Recombinant human binding immunoglobulin protein (BiP) has previously demonstrated anti-inflammatory properties in multiple models of inflammatory arthritis. We investigated whether these immunoregulatory properties could be exploited using gene therapy techniques. A single intraperitoneal injection of lentiviral vector containing the murine BiP (Lenti-mBiP) or green fluorescent protein (Lenti-GFP) transgene was administered in low- or high-dose studies during early arthritis. Disease activity was assessed by visual scoring, histology, serum cytokine and antibody production measured by cell enzyme-linked immunosorbent assay (ELISA) and ELISA, respectively. Lentiviral vector treatment caused significant induction of interferon (IFN)-γ responses regardless of the transgene; however, further specific effects were directly attributable to the BiP transgene. In both studies Lenti-mBiP suppressed clinical arthritis significantly. Histological examination showed that low-dose Lenti-mBiP suppressed inflammatory cell infiltration, cartilage destruction and significantly reduced pathogenic anti-type II collagen (CII) antibodies. Lenti-mBiP treatment caused significant up-regulation of soluble cytotoxic T lymphocyte antigen-4 (sCTLA-4) serum levels and down-regulation of interleukin (IL)-17A production in response to CII cell restimulation. In-vitro studies confirmed that Lenti-mBiP spleen cells could significantly suppress the release of IL-17A from CII primed responder cells following CII restimulation in vitro, and this suppression was associated with increased IL-10 production. Neutralization of CTLA-4 in further co-culture experiments demonstrated inverse regulation of IL-17A production. In conclusion, these data demonstrate proof of principle for the therapeutic potential of systemic lentiviral vector delivery of the BiP transgene leading to immunoregulation of arthritis by induction of soluble CTLA-4 and suppression of IL-17A production.