Developing a barcoded malaria parasite panel to assess broadly neutralizing antibodies

开发带条形码的疟原虫面板来评估广泛中和抗体

基本信息

批准号：
10655645
负责人：
Manoj T Duraisingh
金额：
$ 19.94万
依托单位：
HARVARD SCHOOL OF PUBLIC HEALTH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-06-29 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10655645
关键词：
Affect Alleles Antibodies Antibody-mediated protection Antigen Targeting Antigenic Diversity Antigenic Variation Antigens Antimalarials Area Bar Codes Biological Biological Assay Blood COVID-19 pandemic CRISPR/Cas technology Cells Child Clinical Development Disease Drug Controls Epitopes Equilibrium Erythrocytes Evaluation Exhibits Falciparum Malaria Formulation Genetic Polymorphism Genetic Recombination Genomics Growth High-Throughput Nucleotide Sequencing Human Humoral Immunities Immune Targeting Immunization Immunologics In Vitro Individual Infection Insecticides Invaded Investments Malaria Malaria Vaccines Measurement Measures Mediating Methodology Modification Monoclonal Antibodies Natural Immunity Parasite resistance Parasites Pathogenicity Pathway interactions Phenotype Plasmodium falciparum Population Public Health Recommendation Research Resistance Resources Sampling Specificity Sporozoites Testing Vaccine Research Vaccines Variant World Health Organization burden of illness combat cross immunity functional outcomes immunogenicity inhibiting antibody interest malaria infection neutralizing antibody neutralizing vaccine next generation sequencing novel vaccines parasite invasion pathogen prevent reverse genetics success tool vaccine development vector control

项目摘要

PROJECT SUMMARY Malaria remains a grave threat to global public health with concern that the limited drugs and vector control strategies at our disposal are becoming less effective. Recent progress and renewed interest in new vaccines targeting pathogens, including the Plasmodium falciparum malaria parasite, provide an opportunity to expand our toolkit to combat the disease. A highly efficacious vaccine for malaria, as for other eukaryotic parasites, has been elusive. Substantial effort has been invested in development of a pre-erythrocytic vaccine targeting the sporozoite form of the parasite, resulting in the recent recommendation by the World Health Organization for use in endemic areas. During the clinical blood-stage of malaria, many of the functional antigens that provide the parasite pathways for invasion into erythrocytes are known, and some naturally occurring antibodies are protective. Antigenic diversity and variation, among other factors, poses a major challenge for a broadly- neutralizing blood-stage vaccine, because many immune-targeted molecules for host cell entry are functionally redundant and sequence polymorphic. As antigens continue to be tested for immunogenicity, it will become a high-priority to identify those with the best potential for widespread protection against the breadth of antigenically diverse malaria parasites found across the world. We propose here to develop a platform for facile assessment of the broadly-neutralizing potential of anti-parasite antibodies. In Step 1, we will identify culture-adapted P. falciparum clonal lines that represent all major alleles of the parasite globally; and implement state-of-the-art CRISPR-Cas9 methodologies to introduce short, stably expressed barcode sequences to distinguish the lines. In Step 2, we will pool 90 barcoded lines and validate the use of Illumina-based amplicon-sequencing to deconvolute pools and measure the relative levels of the parasite lines. In Step 3, we will use the pooled assay to measure the strain-specificity of invasion inhibition and broadly-neutralizing potential of monoclonal antibodies targeting two antigens of interest for a blood-stage vaccine. The combination of population genomics, reverse genetic modification of parasites, and quantitative high- throughput sequencing, will permit us to analyze the functional outcomes of antigenic diversity at unprecedented scale and complexity; and will transform efforts to identify the most promising antigens for a potent, broadly- neutralizing malaria vaccine. We further anticipate that the approach will also be useful for non-vaccine related studies for P. falciparum, including analysis of parasite resistance to antimalarials.

项目摘要疟疾仍然是对全球公共卫生的严重威胁，担心有限的药物和媒介控制我们可以使用的策略变得越来越有效。最新进展并重新对新疫苗产生了兴趣靶向病原体，包括恶性疟原虫疟原虫，为扩展提供了机会我们的工具包应对这种疾病。对于其他真核生物寄生虫，一种高效的疟疾疫苗具有难以捉摸。已经投入了大量努力，用于开发针对性疫苗的疫苗寄生虫的孢子岩形式，导致世界卫生组织的最新建议在流行地区。在疟疾的临床血液阶段，许多提供的功能性抗原已知寄生虫入侵红细胞，一些天然存在的抗体是保护的。抗原多样性和变异，除其他因素外，还为广泛的挑战带来了主要挑战。中和血阶段疫苗，因为许多用于宿主细胞进入的免疫靶向分子在功能上都是冗余和序列多态性。随着抗原继续测试免疫原性，它将成为高优先级，以确定具有广泛保护抗原广度的最佳潜力的人世界各地发现的多种疟疾寄生虫。我们在这里提议开发一个便利评估的平台抗寄生虫抗体的广泛中和潜力。在步骤1中，我们将确定适应文化的P。恶性克隆线代表全球寄生虫的所有主要等位基因；并实施最新的 CRISPR-CAS9方法论引入简短，稳定表达的条形码序列以区分线条。在步骤2中，我们将池90条形码线，并验证基于光明的扩增子的使用去声池并测量寄生虫线的相对水平。在步骤3中，我们将使用汇总测定测量单克隆抗体的侵袭抑制和广泛中和潜力的应变特异性针对两种感兴趣的血液疫苗感兴趣的抗原。种群基因组学，寄生虫的反向遗传修饰以及定量高 - 吞吐量测序将使我们能够分析前所未有的抗原多样性的功能结果尺度和复杂性；并将改变努力，以确定有效，广泛的最有希望的抗原中和疟疾疫苗。我们进一步预期该方法也对与非疫苗相关的方法也有用恶性疟原虫的研究，包括分析寄生虫对抗疟疾的耐药性。