ROLE OF Rac AND REACTIVE OXYGEN SPECIES IN KAPOSI'S SARCOMA VIRAL ONCOGENESIS

Rac 和活性氧在卡波西肉瘤病毒癌发生中的作用

• 批准号: 8063961
• 负责人: Pascal J. Goldschmidt-Clermont
• 金额: $ 43.86万
• 依托单位: UNIVERSITY OF MIAMI SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2015-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8063961
• 关键词: Acetylcysteine; Acquired Immunodeficiency Syndrome; Affect; Animal Disease Models; Animal Model; Anti-Retroviral Agents; Antioxidants; Biopsy; Blood; Cell Proliferation; Cells; Chemopreventive Agent; Collaborations; Comprehensive Cancer Center; Data; Development; Down-Regulation; Drug Delivery Systems; Endothelial Cells; FDA approved; Family; Gastrointestinal tract structure; Gender; Gene Expression; Gene Expression Profile; Genes; Genetic; Guanosine Triphosphate Phosphohydrolases; Highly Active Antiretroviral Therapy; Human Herpesvirus 8; Human Identifications; In Vitro; Kaposi Sarcoma; Laboratories; Lead; Lesion; Ligands; Link; Local Therapy; Lung; Malignant Neoplasms; Mediator of activation protein; Modeling; Morbidity - disease rate; Mucous Membrane; Multifocal Lesion; Mus; Mutation; NADPH Oxidase; Names; Oxidative Stress; Pathogenesis; Patients; Pharmaceutical Preparations; Platelet-Derived Growth Factor; Platelet-Derived Growth Factor Receptor; Prevention; Production; Public Health; Reactive Oxygen Species; Role; Signal Transduction; Skin; Smooth Muscle Actin Staining Method; Testing; Transgenic Mice; Universities; Up-Regulation; Vascular Endothelial Growth Factors; Viral; Viral Genes; Viscera; Work; angiogenesis; base; c-myc Genes; cancer type; carcinogenesis; cell transformation; chemotherapy; drinking water; efficacy testing; global health; in vivo; insight; latent gene expression; male; medical schools; member; mortality; mouse model; mutant; new therapeutic target; prevent; promoter; public health relevance; therapeutic target; therapy design; tumor; tumorigenesis

DESCRIPTION (provided by applicant): Kaposi's sarcoma (KS), caused by the Kaposi's sarcoma-associated herpes virus (KSHV), is a major cancer associated with AIDS and a global health challenge. The tumor is characterized by intense angiogenesis and the proliferation of spindle cells that can affect the skin, mucosa and viscera, causing significant morbidity. Understanding the role of viral and cellular genes leading to KS carcinogenesis is paramount to developing rationally designed therapies for KS. A collaboration between the Mesri and Goldschmidt labs has led to the identification of the Rac1 GTPase, a signaling mediator that triggers production of reactive oxygen species (ROS) by non-phagocytic NADPH-oxidase (NOX), as a potential major player in KS. We have found that expression of a constitutively-active Rac1 mutant (RacCA) driven by -smooth muscle actin (-SMA) promoter in transgenic mice led to the formation of lesions that strongly resemble those of Kaposi's sarcoma. Significantly, RacCA--SMA tumors revealed major transcriptome overlap with KS tumor biopsies. RacCA tumorigenesis was linked to male gender, and involved ROS activation of angiogenesis and cell proliferation. Furthermore, we found that AIDS-KS lesions and KSHV-infected tumors from our KS mouse model (mouse endothelial cell KSHV Bac36- mECK36) over-express Rac1 in all KSHV-infected (LANA+ve) cells. Moreover, we found that KS lesions and mECK36 lesions over-express key members of the NOX family and that mECK36 tumors upregulate NOX members in a KSHV dependent fashion. This led us to test the ability of N-acetyl cysteine (NAC), a well characterized antioxidant, to suppress mECK36 tumors in mice. We found that NAC prevented KSHV-induced tumor formation. Interestingly, we also found that NAC inibited VEGF, c-myc and viral gene expression in the mECK36 tumors through a mechanism involving platelet derived growth factor (PDGF) receptor and ligand downregulation. These data indicate that Rac1, NOX, ROS, and their downstream effectors are molecules actively involved in KS viral oncogenesis, and suggest that Rac1 signaling and oxidative stress could be attractive KS chemopreventive and therapeutic targets. We proposse to: Study mechanisms and role of Rac1 activation in KSHV oncogenesis (Aim 1), study the role of NADPH oxidase induction of ROS in KSHV oncogenesis (Aim 2) and to test the efficacy of pharmacologic ROS inhibition on prevention and treatment of RacCA and KSHV-induced tumors (Aim 3). PUBLIC HEALTH RELEVANCE: Kaposi's sarcoma (KS) is the most common type of cancer associated with AIDS (AIDS-KS). KS arises as multifocal lesions in the skin, lungs and gastrointestinal tract characterized by intense blood microvessel and cellular proliferation. Early KS lesions are treated with local and non-toxic therapies; however, advanced KS generally characterized as an advanced disseminated cancer with increased morbidity and mortality is treated with systemic chemotherapy. This treatment is difficult to tolerate for AIDS patients who are receiving Anti Retroviral regimes such as HAART. Thus, the development of rational therapies based on KS pathogenesis is critical to fill this gap. The identification of human and viral genes that are the cause of KS carcinogenesis is important because it could lead to the identification of new therapeutic targets. Essential to this work is the creation of animal models of the disease. Two laboratories of UM/SCCC we have succeeded in generating such a models and identifying a host gene named Rac that by itself can cause in mice many aspects of Kaposi's sarcoma. These models and insights are a very good combination to study mechanisms of Kaposi's sarcoma carcinogenesis. We will study how the Rac gene may participate in cell transformation and tumorigenesis. One of the ways this gene may participate in carcinogenesis is by promoting oxidative stress that can lead to cellular damage and genetic damage that could lead to carcinogenic mutations that may contribute to cancer. So we are going to explore ways of preventing this oxidative genetic damage as a means of preventing and possibly curing Kaposi's sarcoma. So our work will allow us to not only to identify mechanisms of KS carcinogenesis but also will help to identify potential new target and new drugs for anti-KS therapy.

描述（由申请人提供）：卡波西肉瘤（KS），由卡波西肉瘤相关疱疹病毒（KSHV）引起，是一种与艾滋病相关的主要癌症，是全球健康挑战。该肿瘤的特征是强烈的血管生成和梭形细胞的增殖，其可影响皮肤、粘膜和内脏，导致显著的发病率。了解病毒和细胞基因在KS致癌中的作用对于开发合理设计的KS治疗方法至关重要。Mesri实验室和Goldschleman实验室之间的合作导致了Rac 1 GTdR的鉴定，Rac 1 GTdR是一种信号传导介质，可触发非吞噬细胞NADPH氧化酶（NOX）产生活性氧（ROS），作为KS的潜在主要参与者。我们已经发现，在转基因小鼠中由-平滑肌肌动蛋白（-SMA）启动子驱动的组成性活性Rac 1突变体（RacCA）的表达导致了与卡波西肉瘤非常相似的病变的形成。值得注意的是，RacCA-SMA肿瘤显示与KS肿瘤活检的主要转录组重叠。RacCA肿瘤发生与男性有关，并涉及ROS激活血管生成和细胞增殖。此外，我们发现，AIDS-KS病变和KSHV感染的肿瘤从我们的KS小鼠模型（小鼠内皮细胞KSHV Bac 36-mECK 36）过表达Rac 1在所有KSHV感染（拉娜+ve）细胞。此外，我们发现KS病变和mECK 36病变过表达NOX家族的关键成员，并且mECK 36肿瘤以KSHV依赖性方式上调NOX成员。这使我们测试了N-乙酰半胱氨酸（NAC）（一种充分表征的抗氧化剂）抑制小鼠mECK 36肿瘤的能力。我们发现NAC阻止了KSHV诱导的肿瘤形成。有趣的是，我们还发现NAC通过涉及血小板衍生生长因子（PDGF）受体和配体下调的机制抑制mECK 36肿瘤中的VEGF、c-myc和病毒基因表达。这些数据表明，Rac 1，NOX，ROS，和他们的下游效应分子积极参与KS病毒肿瘤发生，并建议Rac 1信号和氧化应激可能是有吸引力的KS化学预防和治疗的目标。我们提议：研究Rac 1激活在KSHV肿瘤发生中的机制和作用（目标1），研究NADPH氧化酶诱导ROS在KSHV肿瘤发生中的作用（目标2），并检测药物ROS抑制对预防和治疗RacCA和KSHV诱导的肿瘤的疗效（目标3）。 公共卫生相关性：卡波西肉瘤（KS）是与艾滋病相关的最常见癌症类型（AIDS-KS）。KS作为皮肤、肺和胃肠道中的多灶性病变出现，其特征在于强烈的血管和细胞增殖。早期KS病变采用局部和无毒疗法治疗;然而，晚期KS通常以发病率和死亡率增加的晚期播散性癌症为特征，采用全身化疗治疗。这种治疗对于正在接受抗逆转录病毒治疗（如HAART）的艾滋病患者难以耐受。因此，开发基于KS发病机制的合理治疗方法是填补这一空白的关键。鉴定人类和病毒基因是KS致癌的原因是重要的，因为它可以导致新的治疗靶点的鉴定。这项工作的关键是建立这种疾病的动物模型。UM/SCCC的两个实验室已经成功地产生了这样的模型，并确定了一个名为Rac的宿主基因，它本身可以在小鼠中引起卡波西肉瘤的许多方面。这些模型和见解是一个很好的结合，研究卡波西肉瘤致癌机制。我们将研究Rac基因如何参与细胞转化和肿瘤发生。这种基因可能参与致癌作用的方式之一是促进氧化应激，这可能导致细胞损伤和遗传损伤，从而导致可能导致癌症的致癌突变。因此，我们将探索预防这种氧化遗传损伤的方法，作为预防并可能治愈卡波西肉瘤的一种手段。因此，我们的工作不仅将使我们能够确定KS致癌机制，而且将有助于确定潜在的新靶点和新的抗KS治疗药物。