Harvard-MIT Department of Health Sciences and Technology, Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Boston, MA 02142, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA; Koch Institute for Integrative Cancer Research, Boston, MA 02142, USA.
Harvard-MIT Department of Health Sciences and Technology, Institute for Medical Engineering and Science, Massachusetts Institute of Technology, Boston, MA 02142, USA; Koch Institute for Integrative Cancer Research, Boston, MA 02142, USA.
Cell Host Microbe. 2018 Mar 14;23(3):395-406.e4. doi: 10.1016/j.chom.2018.01.002. Epub 2018 Feb 22.
The unique relapsing nature of Plasmodium vivax infection is a major barrier to malaria eradication. Upon infection, dormant liver-stage forms, hypnozoites, linger for weeks to months and then relapse to cause recurrent blood-stage infection. Very little is known about hypnozoite biology; definitive biomarkers are lacking and in vitro platforms that support phenotypic studies are needed. Here, we recapitulate the entire liver stage of P. vivax in vitro, using a multiwell format that incorporates micropatterned primary human hepatocyte co-cultures (MPCCs). MPCCs feature key aspects of P. vivax biology, including establishment of persistent small forms and growing schizonts, merosome release, and subsequent infection of reticulocytes. We find that the small forms exhibit previously described hallmarks of hypnozoites, and we pilot MPCCs as a tool for testing candidate anti-hypnozoite drugs. Finally, we employ a hybrid capture strategy and RNA sequencing to describe the hypnozoite transcriptome and gain insight into its biology.
间日疟原虫感染独特的复发性质是消除疟疾的主要障碍。在感染后，休眠的肝期形式，休眠子，潜伏数周至数月，然后复发导致复发性血期感染。关于休眠子生物学知之甚少；缺乏明确的生物标志物，需要支持表型研究的体外平台。在这里，我们使用一种多井格式，结合微图案化原代人肝细胞共培养物（MPCCs），在体外重现间日疟原虫的整个肝期。MPCCs 具有间日疟原虫生物学的关键方面，包括建立持续的小形式和生长的裂殖子、merosome 释放以及随后对网织红细胞的感染。我们发现小形式表现出休眠子的先前描述的特征，并且我们将 MPCCs 作为测试候选抗休眠子药物的工具。最后，我们采用杂交捕获策略和 RNA 测序来描述休眠子转录组，并深入了解其生物学。
