Vaginal microbiota transplant to promote Lactobacillus-dominant cervicovaginal communities

阴道微生物群移植促进以乳杆菌为主的宫颈阴道群落

• 批准号: 10366355
• 负责人: Caroline M Mitchell
• 金额: $ 83.96万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-20 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10366355
• 关键词: 16S ribosomal RNA sequencing; Adverse event; Anaerobic Bacteria; Animal Model; Antibiotic Therapy; Antibiotics; Area; Bacterial Vaginosis; Biological Response Modifier Therapy; Cervical dysplasia; Characteristics; Chemicals; Clinical; Clostridium difficile; Collection; Communities; Complex; Data; Databases; Development; Donor person; Dose; Engraftment; Enrollment; Experimental Models; Formulation; Future; Genes; Genetic; Genital; Genitalia; Goals; HIV; Health; Human; Human Papillomavirus; In Vitro; Individual; Infection; Inflammation; Intervention; Intervention Studies; Kinetics; Lactobacillus; Light; Link; Metabolic; Metronidazole; Mucosal Immune Responses; Mucositis; Oral; Organism; Outcome; Placebos; Premature Birth; Premenopause; Prevalence; Prevention; Probiotics; Public Health; Randomized; Randomized Clinical Trials; Recording of previous events; Recurrence; Relapse; Reproductive Health; Risk; Safety; Saline; Shotguns; Spontaneous abortion; Testing; Transplant Recipients; Transplantation; Treatment Efficacy; Vagina; Woman; Women' s Health; adverse event monitoring; adverse pregnancy outcome; arm; candidate selection; cervicovaginal; comparative genomics; deep sequencing; design; dysbiosis; efficacy evaluation; fecal transplantation; follow-up; genital microbiome; genome sequencing; gut microbiota; immune activation; improved; in vivo; infection risk; inflammatory marker; insight; interest; metabolic profile; metabolomics; metagenomic sequencing; microbial; microbial colonization; microbial community; microbiome; microbiota; microbiota transplantation; multi-component intervention; novel; novel strategies; post intervention; prebiotics; primary outcome; randomized trial; recruit; restoration; safety testing; scale up; secondary outcome; success; therapy design; treatment strategy; vaginal fluid; vaginal lactobacilli; vaginal microbiome; vaginal microbiota; whole genome

PROJECT SUMMARY Vaginal colonization with a Lactobacillus-dominant microbial community is associated with lower risk for preterm birth, HIV acquisition, HPV persistence and cervical dysplasia. A diverse, Lactobacillus-deficient vaginal microbiota such as that seen in bacterial vaginosis (BV) is associated with mucosal inflammation, which is likely the mechanistic link to the adverse health outcomes seen with dysbiosis. Antibiotic treatment for BV achieves short term cure, but recurrence is common. Probiotic treatment to restore healthy lactobacilli is only moderately successful, even with daily treatment. These results emphasize the need for novel strategies to manipulate the genital microbiome and produce sustainable shifts away from dysbiosis. We propose a randomized clinical trial of vaginal microbiota transplant (VMT) with extensive characterization of donors, transplant material, recipients and engraftment, to identify determinants of vaginal microbial colonization and stability. Our team has already obtained an IND, successfully recruited donors, and generated preliminary data demonstrating stability of Lactobacillus in donated material. In Aim 1 we will enroll up to 25 healthy donors and 126 recipients with a history of recurrent BV and an abnormal Nugent score. Recipients receive 1 week of oral metronidazole and are randomized to VMT or saline placebo, two doses given on non-consecutive days in a single week. The primary outcome is prevalence of a Lactobacillus-dominant vaginal microbiota by 16S rRNA sequencing 1 month after intervention. Secondary outcomes include adverse events, Lactobacillus dominance over the entire 6 month follow up, and prevalence of BV by Nugent score at 1, 3 & 6 months. In Aim 2 we will characterize the impact of vaginal fluid transplantation on recipient microbiome (Aim 2.1) and mucosal inflammation (Aim 2.2). For Aim 2.1 we will use 16S rRNA sequencing, qPCR and shotgun metagenomic sequencing (SMS) to define the kinetics of Lactobacillus colonization and community diversity over the 6 months following study intervention. In Aim 2.2 we will assess the impact of VMT vs. placebo on soluble markers of inflammation in vaginal fluid and endocervical immune cell activation. In Aim 3 we will identify genetic characteristics of both successful donations and Lactobacillus isolates cultivated from successful VMT donors and recipients (Aim 3.1). We will compare isolates in vitro to test functional metrics for future selection of strains for novel products (Aim 3.2). We will also compare metabolic profiles of successful vs. unsuccessful donations (Aim 3.3). The resulting isolate collection and database of genes and metabolites associated with achieving Lactobacillus dominance will provide a basis for design of a novel live biotherapeutic for prevention of BV and its associated sequelae. Execution of these three aims will also provide novel insights into determinants of vaginal Lactobacillus colonization and the causal relationship between Lactobacillus and protection from BV, moving the field forward whether or not our trial demonstrates a clinical benefit for VMT. Data obtained will provide guidance on what features are important for designing a synthetic intervention, which would be easier to scale up for widespread use than VMT.

项目摘要 阴道定植以乳酸杆菌为主的微生物群落与早产风险降低相关 出生、艾滋病毒感染、HPV持续存在和宫颈发育不良。一种多样的，缺乏乳酸杆菌的阴道 细菌性阴道病（BV）中的微生物群与粘膜炎症有关，这可能是 生态失调与不良健康结果的机械联系。BV的抗生素治疗 短期治愈，但复发是常见的。益生菌治疗恢复健康的乳酸杆菌只是适度的 即使是日常治疗也是成功的。这些结果强调了需要新的策略来操纵 生殖器微生物组，并产生可持续的转变，远离生态失调。我们提出了一个随机临床 阴道微生物群移植（VMT）试验，对供体，移植材料， 受体和植入，以确定阴道微生物定植和稳定性的决定因素。我们的团队已被 已经获得IND，成功招募了供体，并生成了证明稳定性的初步数据 乳酸杆菌的含量在目标1中，我们将招募多达25名健康供体和126名接受者， BV复发史和Nugent评分异常。接受者接受1周的口服甲硝唑， 随机分配至VMT或生理盐水安慰剂组，在一周内非连续给药两次。主 结果是在治疗后1个月，通过16S rRNA测序发现乳酸杆菌占优势的阴道微生物群的患病率 干预次要结局包括不良事件，整个6个月内乳酸菌占主导地位 随访1个月、3个月和6个月时，根据Nugent评分评估BV患病率。在目标2中，我们将描述 阴道液移植对受体微生物组（Aim 2.1）和粘膜炎症（Aim 2.2）的影响。对于Aim 2.1 我们将使用16S rRNA测序，qPCR和鸟枪宏基因组测序（SMS）来确定 研究干预后6个月内的乳酸菌定植和群落多样性。在目标2.2中，我们 将评估VMT与安慰剂对阴道液和宫颈内可溶性炎症标志物的影响 免疫细胞激活。在目标3中，我们将确定成功捐赠的遗传特征， 从成功的VMT供体和受体中培养的乳杆菌分离株（目的3.1）。我们将比较分离物 在体外测试功能指标，用于未来选择新产品的菌株（目标3.2）。我们还将比较 成功与不成功献血的代谢特征（目标3.3）。所产生的分离株收集和 与实现乳酸杆菌优势相关的基因和代谢物数据库将为 设计一种新的用于预防BV及其相关后遗症的活生物制剂。处决这三个人 aims还将为阴道乳杆菌定植的决定因素和致病因素提供新的见解， 乳酸杆菌和BV保护之间的关系，无论我们的试验是否 证明了VMT的临床获益。所获得的数据将为以下方面提供指导： 设计一种综合干预措施，这将比VMT更容易扩大规模以广泛使用。