转录因子ApiAP2在恶性疟原虫对青蒿素抗性中的作用机制研究

疟原虫对青蒿素的抗性给全球疟疾防控工作带来巨大的挑战，已有研究发现其抗性与K13基因突变有关，但仍存在诸多疑问。有K13突变的“抗性株”大部分仍能被青蒿素杀死，而现场发现没有K13突变的虫株对青蒿素也有耐药。本课题组在前期研究中也发现，恶性疟原虫抗性株在实验室多次传代培养后对青蒿素抗性水平逐步下降，对转录水平有显著差异的转录因子ApiAP2家族成员进行敲除后，对青蒿素敏感性下降。因此我们推测：除K1突变外，恶性疟原虫的青蒿素抗性机制应该还存在非K13依赖的表观遗传学调控途径，而ApiAP2可能是该途径中的关键转录调控因子。本研究拟利用高效的恶性疟原虫CRISPR/Cas9基因编辑技术，并结合转录组、蛋白质-核酸互作和深度测序等技术，进一步验证青蒿素抗性相关AP2家族成员，并分析其调控的青蒿素抗性相关信号通路，从而阐明恶性疟原虫对青蒿素抗性的新机制，为疟原虫青蒿素抗性的监测和应对提供科学依据

Artemisinin resistance of Plasmodium falciparum posed great challenge to global malaria control and elimination. It has been found that the resistance is related to the mutation of K13 gene, but there are still many questions remained unknown. Most parasites of the K13 mutant isolate still can be killed by artemisinin, meanwhile the field studies in Africa found that some isolates without K13 mutant were resistant to artemisinin. Our group also found that the artemisinin resistance level of P. falciparum decreased gradually after repeated passages in the laboratory in vitro culture. Transcriptome analysis revealed that the genes with different transcriptional levels included transcription factors ApiAP2 family members, and artemisinin resistance level of P. falciparum increased after one of PfAp2 knocked out. Therefore, we speculate that artemisinin resistance of P. falciparum is not only related to the mutation of K13, but may also be related to epigenetic pathway, and transcription factors AP2 could be the key player. This study intends to further identify the ApiAP2 family members of P. falciparum, and analysis the pathway related to artemisinin resistance, using CRISPR/Cas9, transcriptomic, protein-nucleic acid interaction, and deep sequencing technology, to elucidate the new mechanism of P. falciparum resistance to artemisinin, providing scientific evidence for surveillance and response of artemisinin resistance of malaria parasite.