Novel approaches to propagate molluscum contagiosum virus in cell culture

在细胞培养中繁殖传染性软疣病毒的新方法

• 批准号: 9089855
• 负责人: Stuart N. Isaacs
• 金额: $ 24.62万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9089855
• 关键词: 3-Dimensional; Adult; Antiviral Response; Attenuated; Benign; Biology; Cell Communication; Cell Culture System; Cell Culture Techniques; Cell Line; Cell model; Cells; Child; Collection; Communities; Complement; Cultured Cells; Development; Distress; Elements; Environment; Epidermis; Extramural Activities; Family; Funding; Future; Genes; Goals; Growth; HIV; Health; Health Care Costs; Human; Immune; Immunocompromised Host; In Vitro; Individual; Infection; Infectious Skin Diseases; Knowledge; Lesion; Life Cycle Stages; Link; Medical; Modeling; Molluscum Contagiosum; Molluscum contagiosum virus; Office Visits; Orthopoxvirus; Parents; Pathway interactions; Patients; Physiology; Population; Poxviridae; Property; Proteins; Research; Research Proposals; Role; Scientist; Signal Pathway; Signal Transduction; Skin; Skin Tissue; Staging; System; Technical Expertise; Testing; Therapeutic; Tissues; United States National Institutes of Health; Viral; Viral Genome; Virus; Virus Diseases; Virus Replication; Work; base; burden of illness; cellular targeting; cost; emotional distress; gene product; immortalized cell; immunosuppressed; in vivo; insight; keratinocyte; novel strategies; pathogen; skin barrier; skin disorder; skin disorder diagnosis; skin lesion; therapeutic development; transmission process; two-dimensional

 DESCRIPTION (provided by applicant): Molluscum contagiosum (MC) is a common skin infection caused by molluscum contagiosum virus (MoCV), a dermatotropic poxvirus. MC presents as single to multiple skin lesions that last from months to years. Transmission is by direct skin-to-skin contact or by indirect contact with fomites. MC lesions are solely restricted t human skin and occur in children, adults, and immunosuppressed individuals. While the lesions are benign, they are often unsightly and thus are psychologically distressful to patients and parents. MC occurs in 2-10% of the worldwide population. In the USA, it constitutes about 1% of all diagnosed skin disorders and occurs in almost 5% of children. In immune-compromised individuals, MC is severe and protracted. Between 5- 18% of HIV patients have MC. Despite the huge burden of disease and the associated significant healthcare costs, there are no extramural NIH-funded projects that specifically study this virus. The major impediment in researching MoCV and finding potential therapeutic cures for MC has been the inability of MoCV to propagate in any type of cultured cells. The objective of this R21 project is to use various novel strategies to identify a system that would allow replication of MoCV in cell culture. We hypothesize that the inability to propagate MoCV in cells in culture is linked to the inability of MoCV to downregulate a cell's innate anti-viral responses. It also reflects unique properties of the human epidermis in vivo that are not reproduced during culture in vitro. We hypothesize that this block in productive infection can be overcome by manipulation of cells in culture to create an intracellular environment that would allow the full MoCV life cycle to be carried out. By establishing models for the growth of MoCV in cell culture and identifying the factors that regulate its replication in keratinocytes, we will gain insights into unique features of skin cells that allow dermatotropic viruses like MoCV to grow and spread in the skin, which will, in the future, serve as a basis for further study and potential development of therapeutics. To test these concepts we will: * Determine at what stage MoCV infection is blocked in both primary human keratinocytes and immortalized cells * Manipulate the host cell microenvironment with poxviral proteins not expressed by MoCV * Manipulate the host cell innate-antiviral responses by targeting cellular proteins By combining Penn's on-campus research expertise in poxvirology and human epidermal biology, we believe that we are poised to make a major advance in poxvirology and discover a way to allow the propagation and spread of MoCV in cell culture systems. Support through this R21 application will allow us begin the steps to reach this important and significant goal.

 描述（由适用提供）：小软体动物（MC）是由皮肤病病毒（MOCV）（MOCV）引起的常见皮肤感染。 MC表现为持续数月到几年的单身至多种皮肤病变。传播是通过直接皮肤接触或与人们间接接触的传播。 MC病变仅受到人类皮肤的限制，发生在儿童，成人和免疫抑制的个体中。虽然病变是良性的，但它们通常是难看的，因此在心理上对患者和父母感到痛苦。 MC发生在2-10％的全球人口中。在美国，它占所有被诊断的皮肤疾病的约1％，并且发生在几乎5％的儿童中。在免疫受损的个体中，MC严重且持久。在5-18％的艾滋病毒患者中患有MC。尽管疾病遭受了巨大的烧伤和相关的重大医疗费用，但没有专门研究该病毒的壁外NIH资助的项目。 MOCV研究MOCV和发现潜在的治疗方法的主要障碍是MOCV无法在任何类型的培养细胞中传播。该R21项目的目的是使用各种新型策略来确定可以在细胞培养中复制MOCV的系统。我们假设无法在培养中传播MOCV与MOCV下调细胞的先天抗病毒反应有关。它还反映了体内人类表皮的独特特性，这些特性未在体外培养过程中再现。我们假设，通过操纵培养物中的细胞创造细胞内环境，可以克服生产感染中的这种阻滞，从而可以实现完整的MOCV生命周期。通过建立MOCV在细胞培养中生长的模型，并确定调节其在角质形成细胞中复制的因素，我们将深入了解皮肤细胞的独特特征 这样一来，像MOCV这样的皮肤病病毒能够在皮肤中生长和扩散，这将来将是进一步研究和潜在治疗发展的基础。要测试这些概念，我们将： *确定在主要人角质形成细胞和永生细胞中，MOCV感染在哪个阶段被阻塞 * *通过MOCV来操纵宿主细胞微环境，并通过MOCV表达poxviral蛋白。生物学，我们认为我们被毒死了，可以在痘病毒学方面取得重大进步，并发现一种允许MOCV在细胞培养系统中传播和传播的方法。通过此R21应用程序的支持将使我们开始采取步骤，以实现这一重要目标。