Malaria Parasite Genomics, Development, Drug Resistance, Pathogenesis, and host-parasite interaction

在2022-2023年期间，我们继续利用约氏疟原虫和伯氏疟原虫研究疟疾发病的分子机制和信号转导途径。这是忙碌的一年，专注于修改两份手稿，致力于正在进行的项目，并启动新的研究项目。 我们在几个项目上取得了良好的进展： 1.完成了疟疾所致贫血的分子机制研究。我们向一家期刊提交了一份描述这项工作的手稿。在收到评论者对论文的评论后，我们进行了许多实验来回应评论者的建议和评论。修改后的手稿再次提交。 2.我们发现一些olfr基因的过度表达可以刺激I型干扰素(INF-I)反应，并增加IRF3和TBK1的磷酸化水平。再一次，提交了一份手稿，我们正在进行额外的实验，以回应评审员的评论。 3.我们正在研究一种新的E3泛素连接酶，它可能在寄生虫的发展和耐药性中发挥重要作用。由于参与这个项目的博士后离职，我们不得不在几个月前重新开始这个项目，换了一个新的博士后。 4.我们继续研究在疟疾寄生虫感染中高表达的小鼠Mmp3基因。我们在CBA背景下建立了MMP3基因敲除小鼠，并进行了一些实验，以研究MMP3在宿主对疟疾寄生虫感染的免疫反应中的作用。我们决定在不同的遗传背景下产生第二个MMP3基因敲除(KO)小鼠，并在C57BL/6J背景下获得新的MMP3基因敲除小鼠。目前，我们正试图培育出同源的MMP3 KO小鼠。 5、我们对另一株约氏疟原虫(N67C)的基因组进行了测序和组装，该菌株是N67的同源寄生虫。我们在2021年发表了N67的基因组。我们还对N67和N67C寄生虫的mRNA进行了测序，以改进基因组注释。目标是确定导致这两种寄生虫不同疾病表型的基因。 我们正在进行一个新的项目，研究感染四个约氏疟原虫菌株和伯氏疟原虫Anka后小鼠的代谢物变化。目前正在对数据进行分析，准备发布。我们还在根据代谢图谱的观察结果设计治疗严重疟疾的实验。 7，在另一个新的项目中，我们进行了约氏疟原虫17XNL和N67C的遗传杂交，克隆了寄生虫的后代，感染了小鼠，并从感染的小鼠中提取了RNA进行测序。我们想要定位和识别在感染了这两种寄生虫的小鼠中导致贫血的寄生虫基因。已经对感染了28个后代的小鼠的RNA样本进行了测序，目前正在分析数据。 8，我们发起了一个项目，研究寄生虫在卵子和子孢子通过基质和体内移动时所施加的力。我们已经获得了这个项目的一些初步数据。 9，我们与肖宇博士合作，完成了一个名为SHIP1的分子的研究项目，该分子可以通过连接I型干扰素信号和自噬之间的串扰来调节抗疟疾免疫(mBio 2023，e03512-22)。

During the year 2022-2023, we continued to focus on studying the molecular mechanisms of malaria pathogenesis and signaling pathways using Plasmodium yoelii and Plasmodium berghei parasites. It was a busy year focusing on revisions of two manuscripts, working on ongoing projects, and initiating new research projects. We have made good progress in several projects: 1. We have finished a study on the molecular mechanism of malaria-induced anemia. We submitted a manuscript describing the work to a journal. After receiving the reviewers comments on the paper, we performed many experiments to address the suggestions and comments from reviewers. The revised manuscript was submitted again. 2. We have shown that over-expression of some Olfr genes could stimulate type I interferon (INF-I) responses and increase phosphorylation levels of IRF3 and TBK1. Again, a manuscript was submitted, and we are working on additional experiments in response to the reviewers comments. 3. We are working on a new E3 ubiquitin ligase that may play an important role in parasite development and drug resistance. Due to the departure of the postdoc who worked on this project, we had to restart this project with a new postdoc a few months ago. 4. We continued to study the mouse mmp3 gene that was shown to be highly expressed in malaria parasite infections. We have generated an mmp3 knockout mouse in the CBA background and performed some experiments to study the functions of MMP3 in host immune response to malaria parasite infections. We decided to generate a second MMP3 knockout (KO) mouse in a different genetic background and have obtained new MMP3 knockout mice in C57BL/6j background. We are trying to generate homologous MMP3 KO mice currently. 5, We have sequenced and assembled the genome of another P. yoelii strain (N67C), an isogenic parasite of N67. We published the genome of N67 in 2021. We have also sequenced the mRNA from the N67 and N67C parasites to improve genome annotation. The goal is to identify genes that contribute to the different disease phenotypes between the two parasites. 6, We are working on a new project studying metabolite changes in mice after infection with four Plasmodium yoelii strains and P. berghei ANKA. The data are being analyzed for publication. We are also designing experiments to treat severe malaria based on the observations from the metabolic profiling. 7, In another new project, we performed genetic crosses of P. yoelii 17XNL and N67C, cloned parasite progenies, infected mice, and extracted RNAs from infected mice for sequencing. We would like to map and identify parasite genes that cause anemia in mice infected with the two parasites. RNA samples from mice infected with 28 progenies have been sequenced, and data are being analyzed currently. 8, We initiated a project to study the forces exerted by the parasites during ookinete and sporozoite movement through the matrix and in vivo. We have obtained some preliminary data on this project. 9, In collaboration with Dr. Xiao Yu, we finished a project studying a molecule called SHIP1 that could modulate antimalarial immunity by bridging the crosstalk between type I IFN signaling and autophagy (mBio 2023, e03512-22).