Di Maggio Lucia S, Fischer Kerstin, Rosa Bruce A, Yates Devyn, Cho Byoung-Kyu, Lukowski Jessica, Calderon Antonia Zamacona, Son Minsoo, Goo Young Ah, Opoku Nicholas O, Weil Gary J, Mitreva Makedonka, Fischer Peter U
Infectious Diseases Division, Department of Medicine, Washington University School of Medicine, St. Louis, MO 63110, USA.
Mass Spectrometry Technology Access Center at McDonnell Genome Institute, Washington University School of Medicine, St. Louis MO 63110.
bioRxiv. 2024 Oct 17:2024.10.15.618383. doi: 10.1101/2024.10.15.618383.
is the agent of onchocerciasis (river blindness) and targeted by WHO for elimination though mass drug administration with ivermectin. A small percentage of adult worms develop pleomorphic neoplasms (PN) that are positively associated with the frequency of ivermectin treatment. Worms with PN have a lower life expectancy and a better understanding about the proteins expressed in PN, and how PN affect protein expression in different tissues could help to elucidate the mechanisms of macrofilaricidal activity of ivermectin. Within a clinical trial of drug combinations that included ivermectin, we detected 24 (5.6%) females with PN by histology of paraffin embedded nodules. To assess the protein inventory of the neoplasms and to identify proteins that may be associated with tumor development, we used laser capture microdissection and highly sensitive mass spectrometry analysis. Neoplasm tissue from three female worms was analyzed, and compared to normal tissues from the body wall, uterus and intestine from the same worms, and to tissues from three females without PN. The healthy females showed all intact embryogenesis. In PN worms, 151 proteins were detected in the body wall, 215 proteins in the intestine, 47 proteins in the uterus and 1,577 proteins in the neoplasms. Only the uterus of one PN female with some stretched intrauterine microfilariae had an elevated number of proteins (601) detectable, while in the uteri of the healthy females 1,710 proteins were detected. Even in tissues that were not directly affected by PN (intestine, body wall), fewer proteins were detected compared to the corresponding tissue of the healthy controls. Immunolocalization of the calcium binding protein OvDig-1 (OVOC8391) confirmed the detection in PN by mass spectrometry. In conclusion we identified proteins that are potentially linked to the development of PN, and systemic dysregulation of protein expression may contribute to worm mortality.
是盘尾丝虫病（河盲症）的病原体，世界卫生组织旨在通过大规模使用伊维菌素进行药物治疗来消除该病。一小部分成年蠕虫会发展为多形性肿瘤（PN），这与伊维菌素治疗的频率呈正相关。患有PN的蠕虫预期寿命较短，更好地了解PN中表达的蛋白质以及PN如何影响不同组织中的蛋白质表达，有助于阐明伊维菌素的杀成虫活性机制。在一项包含伊维菌素的药物组合临床试验中，我们通过石蜡包埋结节的组织学检测到24只（5.6%）患有PN的雌性蠕虫。为了评估肿瘤的蛋白质组成并鉴定可能与肿瘤发展相关的蛋白质，我们使用了激光捕获显微切割和高灵敏度质谱分析。对三只雌性蠕虫的肿瘤组织进行了分析，并与同一只蠕虫的体壁、子宫和肠道的正常组织以及三只没有PN的雌性蠕虫的组织进行了比较。健康的雌性蠕虫显示出完整的胚胎发育过程。在患有PN的蠕虫中，体壁检测到151种蛋白质，肠道检测到215种蛋白质，子宫检测到47种蛋白质，肿瘤检测到1577种蛋白质。只有一只子宫内有一些伸展的微丝蚴的患有PN的雌性蠕虫的子宫中可检测到的蛋白质数量增加（601种），而健康雌性蠕虫的子宫中检测到1710种蛋白质。即使在没有直接受到PN影响的组织（肠道、体壁）中检测到的蛋白质也比健康对照的相应组织少。钙结合蛋白OvDig-1（OVOC8391）的免疫定位证实了质谱在PN中的检测结果。总之，我们鉴定出了可能与PN发展相关的蛋白质，蛋白质表达的系统性失调可能导致蠕虫死亡。
