A unique approach to identify markers for congenital syphilis and neurosyphilis

识别先天性梅毒和神经梅毒标记物的独特方法

• 批准号: 7812566
• 负责人: Nikhat Parveen
• 金额: $ 36.76万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-15 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7812566
• 关键词: Adherence; Africa; Animal Model; Animals; Antibodies; Area; Bacteria; Binding; Biological Models; Borrelia burgdorferi; Brain; Cell Line; Cells; Central Asia; Chronic; Chronic Disease; Congenital Syphilis; Country; Development; Diagnostic; Disease; Early Diagnosis; Eastern Europe; Epithelial; European; Euthanasia; Evaluation; Exhibits; Fibroblasts; Gene Expression; Genes; Genetic; Genetic Techniques; Genome; Globus Pallidus; Homologous Protein; Human; Image; In Vitro; Infection; Lead; Life; Low Birth Weight Infant; Lyme Disease; Mammalian Cell; Marker Vaccines; Membrane Proteins; Molecular; Mus; Neuraxis; Neurons; Neurosyphilis; Order Spirochaetales; Organ; Organism; Pathogenesis; Phase; Pilot Projects; Placenta; Play; Premature Birth; Prevention; Proteins; Rare Diseases; Recovery; Rodent; Role; Serologic tests; Shuttle Vectors; Site; Specificity; Surface; Syphilis; System; Testing; Time; Tissues; Translational Research; Treponema pallidum; Virulence Factors; abortion; base; experience; extracellular; gain of function; in vivo; innovation; novel diagnostics; novel strategies; novel vaccines; numb protein; optical imaging; pathogen; promoter; protein expression; public health relevance; research study; stillbirth; tool; transmission process; vaccine candidate

DESCRIPTION (provided by applicant): Broad Challenge Area-Translational Science (15) Specific Challenge Topic- 15-OD(ORDR)-101*: Pilot projects for prevention, early detection and treatment of rare diseases, Syphilis is a chronic, multisystemic disease caused by the spirochete, Treponema pallidum subspecies pallidum (T. pallidum). The spirochete is acquired by sexual contact and is prevalent worldwide. Congenital syphilis and neurosyphillis manifestation of the disease have become rare in the Western countries but they remain a problem in underdeveloped countries of Africa, Central Asia and Eastern Europe. Congenital syphilis is devastating and can result in stillbirths, abortions, low birth weights or premature births. Neurosyphillis is also re-emerging around the world. T. pallidum sequence revealed itself to be one of the smallest genomes (1.1Mb) among bacteria with a few potential surface proteins that can determine the spirochete's ability to colonize various tissues during infection. Spirochetes are extracellular pathogens and their adherence to mammalian cells appears to play a critical role in tissue colonization. However, several challenges have resulted in the poor understanding of T. pallidum pathogenesis. (i) T. pallidum cannot be grown in vitro. This limits the ability to investigate this organism thoroughly. (ii) Genetic techniques are not feasible for this spirochete. (iii) T. pallidum exhibits extreme host specificity with humans as its only natural host. Therefore, there is a desperate need to develop novel strategies to investigate the molecular basis of T. pallidum pathogenesis especially relevant to congenital syphilis and neurosyphillis using a small animal model system. T. pallidum is closely related to Borrelia burgdorferi, which causes Lyme disease. Both of these spirochetes are structurally and physiologically similar organisms and share several homologous proteins. T. pallidum and B. burgdorferi exhibit different phases of disease in their hosts, including disseminated infection of various tissues followed by latent and chronic disease. Development of genetic tools for B. burgdorferi in the past decade has led to a significant progress in the understanding of Lyme disease pathogenesis. In this proposal, we will employ a highly innovative and unconventional approach of using bioluminescent B. burgdorferi as a surrogate system to study the role of selected T. pallidum molecules in cell-specific interactions and pathogenesis. Our hypothesis is that expression of one or more T. pallidum proteins in bioluminescent B. burgdorferi will promote invasion and colonization of specific sites. Invasion of the placenta, which may lead to congenital transmission and of the central nervous system will be detectable in mice by an in vivo imaging system (IVIS 200). The following studies will test this hypothesis. Specific Aim 1: To examine the expression and localization of T. pallidum proteins in B. burgdorferi and assess their roles in adherence to specific mammalian cell lines in vitro. Specific Aim 2: To determine if B. burgdorferi acquires the ability to colonize brain and placenta of mice efficiently after expression of T. pallidum protein(s) and also promotes congenital transmission of Lyme spirochetes from placenta to the litter. Significance Development of a new bioluminescent B. burgdorferi surrogate model system in this proposal will help visualize disseminated colonization of the mouse placenta and neuronal tissues facilitated by the expression of T. pallidum proteins. This gain of function approach will help fill the gap in the understanding of this historic, uncultivable spirochete. Furthermore, there are no promising vaccine candidates identified against syphilis to date. We expect that our experience in B. burgdorferi and expertise of our collaborators, Drs. Sheila Lukehart and Arturo Centurion, in T. pallidum pathogenesis will help us achieve our objective to identify critical virulence factors of T. pallidum and determine their potential as diagnostic markers and novel vaccine candidates. PUBLIC HEALTH RELEVANCE: Syphilis is a chronic multisystemic disease and congenital syphilis and neurosyphillis manifestation of the disease are rare in the Western countries. Congenital syphilis is devastating and can result in stillbirths, abortions, low birth weights or premature births and neurosyphillis is also re-emerging around the world. Using bioluminescent bacteria as surrogate system to express uncultivable syphilis-causing spirochete proteins in this proposal will facilitate examination of colonization of mouse placenta and neuronal tissues and will also help in identification of new diagnostic markers and vaccine candidates.

描述（由申请人提供）：广泛的挑战领域 - 跨性科学（15）特定挑战主题-15-OD（ORDR）-101*：预防，早期检测和治疗罕见疾病的试点项目，梅毒是一种由螺旋藻，Treponema pallidum pallidum subspecies pallidum（T。pallidum（T。pallidum）引起的慢性，多系统疾病。螺旋体是通过性接触获得的，并且在全球范围很普遍。该疾病的先天性梅毒和神经淋巴结的表现在西方国家已经很少见，但在非洲，中亚和东欧的欠发达国家中，它们仍然是一个问题。先天性梅毒是毁灭性的，可能导致死产，流产，低出生体重或早产。 Neurosyphillis也在世界各地重新出现。 T. pallidum序列表明自己是细菌中最小的基因组之一（1.1MB），具有一些潜在的表面蛋白质，可以确定螺旋体在感染过程中定居各种组织的能力。螺旋体是细胞外病原体，它们对哺乳动物细胞的粘附似乎在组织定植中起关键作用。然而，一些挑战导致对pallidum发病机理的理解不足。 （i）T。pallidum不能在体外生长。这限制了对这种生物进行彻底研究的能力。 （ii）对于这种螺旋体而言，遗传技术是不可行的。 （iii）T。pallidum与人类作为其唯一的天然宿主表现出极端的宿主特异性。因此，迫切需要制定新的策略来研究使用小型动物模型系统，研究与先天性梅毒和神经淋巴细胞的分子基础。 T. pallidum与引起莱姆病的Borrelia burgdorferi密切相关。这两种螺旋体在结构和生理上相似的生物都具有相似的生物，并共享几种同源蛋白。 Pallidum和B. burgdorferi在其宿主中表现出不同的疾病阶段，包括对各种组织的传播感染，随后是潜在和慢性疾病。在过去的十年中，开发了伯格多菲利芽孢杆菌的遗传工具已导致对莱姆病发病机理的理解取得了重大进展。在此提案中，我们将采用一种高度创新的和非常规的方法，将生物发光的B. burgdorferi用作替代系统来研究选定的pallidum分子在细胞特异性相互作用和发病机理中的作用。我们的假设是，在生物发光的B. burgdorferi中，一种或多种pallidum蛋白的表达将促进特定部位的侵袭和定植。胎盘的侵袭可能导致先天性传播和中枢神经系统的侵袭在小鼠中可以通过体内成像系统检测到（IVIS 200）。以下研究将检验这一假设。具体目的1：检查B. burgdorferi中颗粒蛋白的表达和定位，并评估其在体外遵守特定哺乳动物细胞系中的作用。具体目的2：确定B. burgdorferi是否获得了在表达T. pallidum蛋白（S）表达后有效地定居小鼠的大脑和胎盘的能力，并促进了莱姆螺旋体从胎盘向垃圾到垃圾的先天性传播。在该提案中，新的生物发光B. burgdorferi替代模型系统的显着性发展将有助于可视化小鼠胎盘和神经元组织的传播定殖，并通过T. pallidum蛋白的表达促进。这种功能方法的增益将有助于填补对这种历史性，无法培养的螺旋体的理解。此外，迄今为止，还没有针对梅毒的疫苗候选者。我们希望我们在B. Burgdorferi的经验以及合作者Drs的专业知识。 Sheila Lukehart和Arturo Centurion在T. pallidum发病机理中将有助于我们实现我们的目标，以鉴定Pallidum的关键毒力因子，并确定其作为诊断标记和新型疫苗候选物的潜力。 公共卫生相关性：梅毒是一种慢性多系统疾病，先天性梅毒和神经性疾病的表现在西方国家很少见。先天性梅毒是毁灭性的，可能导致死产，堕胎，低出生体重或早产，而神经淋巴细胞也在世界各地重新出现。在本提案中，使用生物发光细菌作为替代系统来表达不可养殖的梅毒引起的螺旋蛋白，将促进小鼠胎盘和神经元组织的定殖，也将有助于鉴定新的诊断标记物和疫苗候选者。