Deciphering Host–Parasite Interplay in <i>Leishmania</i> Infection through a One Health View of Proteomics Studies on Drug Resistance

Recent efforts in the study of vector-borne parasitic diseases (VBPDs) have emphasized an increased consideration for preventing drug resistance and promoting the environmental safety of drugs, from the beginning of the drug discovery pipeline. The intensive use of the few available antileishmanial drugs has led to the spreading of hyper-resistant Leishmania infantum strains, resulting in a chronic burden of the disease. In the present work, we have investigated the biochemical mechanisms of resistance to antimonials, paromomycin, and miltefosine in three drug-resistant parasitic strains from human clinical isolates, using a whole-cell mass spectrometry proteomics approach. We identified 14 differentially expressed proteins that were validated with their transcripts. Next, we employed functional association networks to identify parasite-specific proteins as potential targets for novel drug discovery studies. We used SeqAPASS analysis to predict susceptibility based on the evolutionary conservation of protein drug targets across species. MATH-domain-containing protein, adenosine triphosphate (ATP)-binding cassette B2, histone H4, calpain-like cysteine peptidase, and trypanothione reductase emerged as top candidates. Overall, this work identifies new biological targets for designing drugs to prevent the development of Leishmania drug resistance, while aligning with One Health principles that emphasize the interconnected health of people, animals, and ecosystems.近年来，针对媒介传播寄生虫病（vector-borne parasitic diseases, VBPDs）的研究愈发强调，需从药物研发管线伊始便兼顾耐药性防控与药物环境安全性。现有抗利什曼病药物的大量使用，导致高耐药性婴儿利什曼原虫（Leishmania infantum）毒株的传播，给该疾病带来了长期的流行负担。本研究针对三株来源于人类临床分离株的耐药寄生虫毒株，采用全细胞质谱蛋白质组学方法，探究了其对锑剂、巴龙霉素与米替福新的耐药生化机制。我们共鉴定出14种差异表达蛋白，并通过转录组数据对其进行了验证。随后，我们借助功能关联网络筛选出可作为新型药物研发潜在靶点的寄生虫特异性蛋白。我们采用SeqAPASS分析，基于蛋白药物靶点在不同物种间的进化保守性预测药物敏感性。含MATH结构域蛋白、三磷酸腺苷（ATP）结合盒B2、组蛋白H4、钙蛋白酶样半胱氨酸肽酶及锥虫硫酮还原酶，成为最具潜力的候选靶点。综上，本研究鉴定出可用于研发预防利什曼原虫耐药性新药的新型生物学靶点，同时契合了强调人类、动物与生态系统健康互联互通的同一健康（One Health）原则。