Gene Therapy for Liver Diseases

Ｍａｎｙｖｅｃｔｏｒｓｈａｖｅｂｅｅｎｓｔｕｄｉｅｄｆｏｒｄｅｌｉｖｅｒｙｏｆｎｕ ｃｌｅｉｃａｃｉｄｓｔｏｔｈｅｌｉｖｅｒ .Ｔｈｅｓｅｗｉｌｌｎｏｔｂｅｒｅｖｉｅｗｅｄｈｅｒｅ .Ｉｔｉｓｃｌｅａｒｔｈａｔｎｏｎｅｏｆｔｈｅｓｅｖｅｃｔｏｒｓａｒｅｉｄｅａｌ.Ｔｏｄａｙ ,ｔｈｉｓｐｒｅｓｅｎｔａｔｉｏｎｗｉｌｌｆｏｃｕｓｏｎｗｏｒｋｏｎｒｅｃｅｐｔｏｒ ｍｅｄｉａｔ ｅｄＤＮＡｄｅｌｉｖｅｒｙ .Ｉｎｔｈｉｓａｐｐｒｏａｃｈ ,ｎａｔｕｒａｌｃｅｌｌ ｓｕｒｆａｃｅｒｅｃｅｐｔｏｒｓａｒｅｕｓｅｄｔｏｉｎｔｅｒｎａｌｉｚｅｎｕｃｌｅｉｃａｃｉｄｓｂｙｃｏｕｐｌｉｎｇａｎａｐｐｒｏｐｒｉａｔｅｌｉｇａｎｄｗｉｔｈｔｈｅＤＮＡｏｆｉｎｔｅｒｅｓｔ .ＴｈｉｓｉｎｖｏｖｌｅｓｔｈｅｆｏｒｍａｔｉｏｎｏｆａｃｏｍｐｌｅｘｂｅｔｗｅｅｎＤＮＡａｎｄａｐｏｌｙｃａｔｉｏｎｗｈｉｃｈｂｅａｒｓａｃｏｖａｌｅｎｔｌｙｌｉｎｋｅｄｌｉｇａｎｄｍｏｉｅｔｙｓｐｅｃｉｆｉｃｆｏｒａｇｉｖｅｎｒｅｃｅｐｔｏｒｏｎｔｈｅｓｕｒｆａｃｅｏｆｃｅｌｌｓｉｎｔｈｅｔａｒｇｅｔｔｉｓｓｕｅ .Ｗｅｈａｖｅｕｓｅｄａｓｉａｌｏｇｌｙｃｏｐ...     

Gene Therapy for Heart Failure: New Perspectives

Purpose of Review

The current knowledge of pathophysiological and molecular mechanisms responsible for the genesis and development of heart failure (HF) is absolutely vast. Nonetheless, the hiatus between experimental findings and therapeutic options remains too deep, while the available pharmacological treatments are mostly seasoned and display limited efficacy. The necessity to identify new, non-pharmacological strategies to target molecular alterations led investigators, already many years ago, to propose gene therapy for HF. Here, we will review some of the strategies proposed over the past years to target major pathogenic mechanisms/factors responsible for severe cardiac injury developing into HF and will provide arguments in favor of the necessity to keep alive research on this topic.

Recent Findings

After decades of preclinical research and phases of enthusiasm and disappointment, clinical trials were finally launched in recent years. The first one to reach phase II and testing gene delivery of sarcoendoplasmic reticulum calcium ATPase did not yield encouraging results; however, other trials are ongoing, more efficient viral vectors are being developed, and promising new potential targets have been identified. For instance, recent research is focused on gene repair, in vivo, to treat heritable forms of HF, while strong experimental evidence indicates that specific microRNAs can be delivered to post-ischemic hearts to induce regeneration, a result that was previously thought possible only by using stem cell therapy.

Summary

Gene therapy for HF is aging, but exciting perspectives are still very open.

     

平滑肌细胞基因治疗血管再狭窄的新进展

血管平滑肌细胞在再狭窄形成的病理过程中发挥了重要作用,现从不同目的基因及转录因子诱骗策略方面总结了近几年来针对平滑肌细胞的基因治疗,重点介绍了诱骗治疗策略的原理、特征、应用及前景,并对载体的选择和输送转染方式进行了回顾,指出了基因治疗领域存在的问题和发展方向。     

白细胞介素12对小鼠肝癌基因治疗的实验研究

目的：研究白细胞介素１２对小鼠肝癌细胞基因治疗效果。方法：利用脑心肌炎病毒（ＥＭＣＶ）及脊髓灰质炎（Ｐｏ－ｌｉｏ）病毒内核糖体进入位点（ＩＲＥＳ）,连接ｍＩＬ－１２ Ｐ４０及ｐ３５ ｃＤＮＡｓ和筛选基因新霉术磷酸转移酶（ＮｅｏＲ）,克隆至逆转录病毒载体ｐＧＣＥＮ中,使三个基因同时受逆转录病毒载体５’端ＬＴＲ启动子控制,转录至同一ｍＲＮＡ转录本上,从而构建成多顺反子逆转录病毒载体,即ｐＧＣＥＮ／ｍＩＬ－１２。在ＬｉｐｏｆｅｃｔＡＭＩＮＥ介导下将ｐＧＣＥＮ／ｍＩＬ－１２转染包装细胞ＰＡ３１７,Ｇ４１８筛选,直至出现阳性克隆（命名为：Ｍ４５／ｍＩＬ－１２）,挑取抗性克隆,扩大培养,收集上清,用小鼠成纤维细胞ＮＩＨ３Ｔ３测定病毒滴度。然后用重组逆转录病毒感染小鼠肝癌细胞ＭＭ４５Ｔ．Ｌｉ,Ｇ４１８筛选,直至出现阳性克隆,扩大培养,对阳性克隆进行鉴定。将６０Ｃｏ照射（６０ Ｇｙ）Ｍ４５／ｍＩＬ－１２细胞对荷瘤小鼠进行瘤内接种,每周一次,连续治疗三次,观察其治疗效果。结果：病毒上清中重组逆转录病毒滴度为５×１０~５ＣＦＵ／ｍｌ。 ＰＣＲ及ＲＴ－ＰＣＲ证明外源基因已整合至小鼠肝癌细胞基因组中,以及外源基因在ｍＲＮＡ水平上的表达。     

高血压病的基因治疗

多年来 ,高血压病的治疗一直依赖于药物 ,但现有的降压药物均存在不同程度的副作用和并发症 ,严重的副作用迫使部分病人放弃治疗 [1]。由于高血压病为终生性疾病 ,有效的降压治疗必须每天服用降压药 ,这也使部分病人难以坚持治疗 [1] 。而且据调查仅有 2 0 %～ 35 %的高血压病人药物治疗取得令人满意的效果[2 ] 。此外 ,对于人体生理功能而言药物均是外来人工合成品 ,从生理角度出发 ,人们更欢迎用生物途径来解决生物问题 ,例如 :基因治疗高血压病。基因治疗具有高度的靶组织特异性和生物活性 ,以及疗效的长久性(数天、数周、数月 ) ,而且副…     

Alphaviruses in Gene Therapy

Alphavirus vectors present an attractive approach for gene therapy applications due to the rapid and simple recombinant virus particle production and their broad range of mammalian host cell transduction. Mainly three types of alphavirus vectors, namely naked RNA, recombinant particles and DNA/RNA layered vectors, have been subjected to preclinical studies with the goal of achieving prophylactic or therapeutic efficacy, particularly in oncology. In this context, immunization with alphavirus vectors has provided protection against challenges with tumor cells. Moreover, alphavirus intratumoral and systemic delivery has demonstrated substantial tumor regression and significant prolonged survival rates in various animal tumor models. Recent discoveries of the strong association of RNA interference and disease have accelerated gene therapy based approaches, where alphavirus-based gene delivery can play an important role.     

心力衰竭的基因治疗

心力衰竭是多种心脏疾病的终末阶段,一般常规治疗预后不佳。近年来不断涌现的基因治疗方法,补充缺失的正常功能蛋白质、抑制体内某些基因的过度表达等,可以从根本上达到治疗心力衰竭的目的,为心力衰竭治疗提供了新的契机。现就心力衰竭基因治疗的作用靶点以及心脏基因转运技术的研究进展等进行综述。     

Maggot Therapy Takes Us Back to the Future of Wound Care: New and Improved Maggot Therapy for the 21st Century

In the 21st century, eighty years after William Baer presented his groundbreaking work treating bone and soft tissue infections with live maggots, thousands of therapists around the globe have rediscovered the benefits of maggot therapy. The renaissance in maggot therapy is due in large part to recent technological advancements that have solved or minimized many of the treatment''s earlier drawbacks: the need for reliable access to this perishable medical device, simplified application, and low-cost production. Modern dressing materials have simplified the procedure and minimized the risk of escaping maggots. The establishment of dozens of laboratories throughout the world, along with access to overnight courier services in many regions, has made medicinal maggots readily available to millions of people in need. Studies show that fears of patient nonacceptance are unfounded. The medical literature is rapidly growing with scientific evidence demonstrating the efficacy and safety of maggot therapy for a variety of problematic wounds. This article examines how these and other technologies are optimizing the study and application of maggot therapy for wound care.     

原发性肝癌放射治疗进展

原发性肝癌（PLC）是常见的恶性肿瘤之一，以往主要的治疗方法是手术切除。随着新的放疗技术，如三维适形放射治疗、立体定向放射治疗、束流调强放射治疗等的发展，放射治疗在PLC综合治疗中的作用日益突出。本文对PLC放射治疗的进展，包括PLC放疗的理论基础、放疗技术的发展、放疗与其他方法的联合应用、放疗并发症的发生和处理作一概述。     

Heart Failure Therapy at the Turn of the Century

Major changes in the treatment of heart failure have occurred in the last fifty years that have had a dramatic effect on its morbidity and mortality. Over two hundred years have passed since the demonstration of the benefit of digitalis in heart failure to the development of potent loop diuretics. The observation that vasodilators could improve both cardiac function and mortality led to the investigation of the Angiotensin Converting Enzyme Inhibitors (ACEI). Although these agents had significant vasodilator properties, their major benefit appears to be related to their ability to effect remodeling of the failing left ventricle. The most recent randomized clinical trials demonstrate that Beta Adrenergic Blocking agents can provide an incremental effect on both mortality and morbidity when added to therapy with ACEI. Although these agents have improved the outlook for the heart failure patient, they have had very little effect on the improvement of left ventricular function. Future research must be directed at methods to deal with this issue by either changing the contractile properties of the cardiomyocyte by pharmacologic or electrical therapy or by transplanting functional cells that can increase the number of functioning contractile units.     

Cell-Type Specific Interferon Stimulated Gene Staining in Liver Underlies Response to Interferon Therapy in Chronic HBV Infected Patients

Background

Interferon-α is approved as one of the main therapeutic treatments for chronic hepatitis B virus infection, but only a small number of patients achieve sustained virological response. The molecular mechanisms underlying IFN-α resistance in those patients who do not respond remain elusive. Previous work in our laboratory identified the pre-activation of IFN signaling leading to increased expression of a subset of interferon stimulated genes in the pretreatment liver tissues of chronic HBV infected patients correlated with treatment non-response.

Aims

We studied the cell-type specific gene expression of interferon stimulated genes in the liver of chronic HBV infected patients and the cellular basis of the phenotype through ISG15 and MxA protein expression.

Methods

Immunohistochemical analysis was used to detect the expression of ISG15 and MxA protein in the pretreatment liver tissues of chronic HBV infected patients and the expression patterns were correlated with treatment outcomes.

Results

In the non-responders, ISG15 and MxA protein expression in the pretreatment liver tissues was more pronounced in hepatocytes while in the responders, ISG15 and MxA protein expression was more focused in macrophages. ISG15 and MxA proteins were occasionally expressed in hepatocytes in normal livers.

Conclusion

There were significant differences in the cell-type specific protein expression of ISG15 and MxA in the pretreatment liver tissues of chronic HBV infected patients between treatment responders and non-responders. An easy prediction method based on immunohistochemical stains of a subset of interferon stimulated genes may be developed to predict treatment outcomes of IFN therapy in chronic HBV infected patients.     

心肌血管新生的基因治疗

尽管冠状动脉粥样硬化性心脏病的防治取得了很大进步 ,并广泛开展了冠状动脉成形术和冠状动脉搭桥术 ,但仍有许多冠心病患者未能获得有效治疗。为了改善缺血心肌的血流 ,分子生物学领域的研究已成为当今的热门 ,用基因转移技术促进缺血心肌组织的血管新生也将成为一种极有前途的治疗方法。1　基因治疗的背景研究用转基因技术治疗缺血心肌的实验背景主要来源于对大量重组的促血管生长因子的研究。 2 0多年前 ,Folkman首先提出血管生长因子的治疗作用 ,这种观点是基于对肿瘤的研究 ,人们发现肿瘤的发展 ,依赖于血管的新生 ,与这些肿瘤相关的…     

Gene Transfer Therapy in Vascular Diseases

Somatic gene therapy of vascular diseases is a promising new field in modern medicine. Recent advancements in gene transfer technology have greatly evolved our understanding of the pathophysiologic role of candidate disease genes. With this knowledge, the expression of selective gene products provides the means to test the therapeutic use of gene therapy in a multitude of medical conditions. In addition, with the completion of genome sequencing programs, gene transfer can be used also to study the biologic function of novel genes in vivo. Novel genes are delivered to targeted tissue via several different vehicles. These vectors include adenoviruses, retroviruses, plasmids, plasmid/liposomes, and oligonucleotides. However, each one of these vectors has inherent limitations. Further investigations into developing delivery systems that not only allow for efficient, targeted gene transfer, but also are stable and nonimmunogenic, will optimize the clinical application of gene therapy in vascular diseases. This review further discusses the available mode of gene delivery and examines six major areas in vascular gene therapy, namely prevention of restenosis, thrombosis, hypertension, atherosclerosis, peripheral vascular disease in congestive heart failure, and ischemia. Although we highlight some of the recent advances in the use of gene therapy in treating vascular disease discovered primarily during the past two years, many excellent studies published during that period are not included in this review due to space limitations. The following is a selective review of practical uses of gene transfer therapy in vascular diseases. This review primarily covers work performed in the last 2 years. For earlier work, the reader may refer to several excellent review articles. For instance, Belalcazer et al. (6) reviewed general aspects of somatic gene therapy and the different vehicles used for the delivery of therapeutic genes. Gene therapy in restenosis and stimulation of angiogenesis in the cardiac muscle are discussed in reviews by several investigators (13,26,57, 74,83). In another review, Meyerson et al. (43) discuss advances in gene therapy for vascular proliferative disorders and chronic peripheral and cardiac ischemia.     

缺血性心血管疾病的基因治疗研究进展

基因治疗是缺血性心血管疾病有希望的治疗方法之一,目前缺血性心血管疾病基因治疗的靶点多为内皮细胞、平滑肌细胞、心肌细胞以及与他们有关的细胞和组织等。基因治疗的方向主要包括:治疗冠状动脉的病变、作为其它治疗的辅助治疗手段、缓解或治疗血管疾病导致的并发症。具体措施是:靶向治疗血管病变、治疗性血管生成、心肌保护性治疗、细胞或组织的再生与修复、靶向致病基因的治疗、靶向细胞周期的治疗、靶向治疗导致心血管病变的原发病、冠状动脉再狭窄的预防与治疗等。     

Extracorporeal Liver Support Therapy With Prometheus in Patients With Liver Failure in the Intensive Care Unit

Acute liver failure (ALF) and acute‐on‐chronic liver failure (AoCLF) are associated with a high mortality. In these patients an accumulation of both water‐soluble and water‐insoluble, protein‐bound, metabolic waste products occurs. Conventional extracorporeal blood purification techniques based on diffusion and/or convection such as hemodialysis or hemofiltration may only eliminate small molecular weight, water‐soluble compounds. In recent years, fractionated plasma separation and adsorption (FPSA) with the Prometheus system has been introduced for extracorporeal liver support therapy. To date, however, only limited data is available regarding the effect of this treatment on mortality and outcome of patients with advanced liver disease. Here we report on our experience with 23 patients with severe liver failure who were treated with Prometheus in our medical intensive care unit. Fourteen patients had AoCLF, and nine patients experienced ALF. The median bilirubin level at the start of Prometheus therapy was 30.5 mg/dL and the median Acute Physiology and Chronic Health Evaluation II (APACHE II) score was 26. During 40 individual treatment sessions lasting 5–6 h, Prometheus therapy reduced serum bilirubin levels from 23.7 mg/dL to 15.0 mg/dL (median values) (P < 0.001), and the overall survival was 26%. ALF patients had a better survival compared to AoCLF patients (44% vs. 22%; P = 0.022). Apart from one patient who developed hemodynamic instability during a treatment session, Prometheus therapy was well tolerated without relevant side‐effects. In conclusion, extracorporeal liver support therapy with Prometheus is a novel and safe treatment option in patients with severe liver failure. In this series, patients with ALF showed a significantly better outcome with Prometheus therapy compared to AoCLF patients.