A novel Approach to Target Trypanosome, Leishmania and Superbugs to the Lysosomal Compartment

一种将锥虫、利什曼原虫和超级细菌靶向溶酶体区室的新方法

• 批准号: RGPIN-2017-03896
• 负责人: Morales, Carlos
• 金额: $ 2.04万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=710601
• 关键词: novel; Approach; Target; Trypanosome; Leishmania

The long term objective of this research is to develop a novel therapeutic approach that will revolutionize the way transmissible diseases are treated, by offering an alternative therapy to antiviral drugs, antibiotics and vaccines. Trypanosomiasis (Trypanosoma spp.), leishmaniosis (Leishmania spp.), brucellosis (Brucella spp.), and tuberculosis (Mycobacterium spp.), for which there are no effective cures or treatments are major infectious agents in wild species, domestic animals and humans. It is well known that the majority of pathogens invade the cell via endocytosis, by attaching to existing cell surface receptors. Once inside the cell, the pathogens trick the phagosomes and lysosomes and preferentially divide within them. In eukaryotic cells the delivery of newly synthesized proteins to endosomes, phagosomes and lysosomes is dependent on the Golgi apparatus, i.e., the organelle that sorts and targets proteins to these destinations. Proteins delivered to endosomes, phagosomes and lysosomes include enzymes and activator proteins. They are directed therein via special sorting receptors. We found that a region in the lysosomal activator protein “Prosaposin” (PSAP) contains a motif required for its binding to a sorting receptor called “Sortilin” and consequently for its transport to the endolysosomal compartment. During our last “NSERC Grant” we demonstrated that the antimicrobial proteins -Defensin and Tachyplesin linked to the lysosomal targeting motif of PSAP were delivered to endosomes and lysosomes. In this proposal, we would like to test the following hypotheses: 1) -Defensin-PSAP and Tachyplesin-PSAP are directed to the endolysosomal system via Sortilin; 2)Tachyplesin-PSAP has trypanocidal and leishmanocidal effects in the lysosomal compartment of infected cells; 3) -Defensin-PSAP has a bactericidal effect in the lysosomal compartment of infected cells. Thus, the present study will assess the effectiveness of these antimicrobial fusion proteins to kill trypanosomes, leishmania and bacteria within phagosomes and lysosomes. Our study will permit to establish a proof of concept of a new strategy for the elimination of pathogens in the endo/lysosomal compartment. The outcome of our research will be relevant for the treatment of infectious diseases in wild, endangered, and domestic animal species, as well as in humans and will open innumerable avenues to examine the delivery and effects of antipathogenic peptides in viral, bacterial and protozoal infections.

这项研究的长期目标是开发一种新颖的治疗方法，通过提供抗病毒药物、抗生素和疫苗的替代疗法，彻底改变传染性疾病的治疗方式。锥虫病（Trypanosoma spp.）、利什曼病（Leishmania spp.）、布鲁氏菌病（Brucella spp.）和结核病（分枝杆菌属 spp.）尚无有效的治愈或治疗方法，是野生物种、家畜和人类的主要传染源。众所周知，大多数病原体通过附着于现有的细胞表面受体，通过内吞作用侵入细胞。一旦进入细胞，病原体就会欺骗吞噬体和溶酶体，并优先在其中分裂。在真核细胞中，新合成的蛋白质向内体、吞噬体和溶酶体的递送依赖于高尔基体，即对蛋白质进行分类和靶向这些目的地的细胞器。递送至内体、吞噬体和溶酶体的蛋白质包括酶和激活蛋白。它们通过特殊的分选受体被引导至其中。我们发现溶酶体激活蛋白“Prosaposin”（PSAP）中的一个区域包含其与称为“Sortilin”的分选受体结合所需的基序，从而将其转运至内溶酶体区室。在我们上次的“NSERC 资助”期间，我们证明了与 PSAP 的溶酶体靶向基序相连的抗菌蛋白 - 防御素和鲎素被递送到内体和溶酶体。在本提案中，我们想测试以下假设： 1) -Defensin-PSAP 和 Tachyplesin-PSAP 通过 Sortilin 定向至内溶酶体系统； 2) Tachyplesin-PSAP 在感染细胞的溶酶体区室中具有杀锥虫和杀利什曼病的作用； 3) -Defensin-PSAP 在受感染细胞的溶酶体区室中具有杀菌作用。因此，本研究将评估这些抗菌融合蛋白杀死锥虫、利什曼原虫以及吞噬体和溶酶体内细菌的有效性。我们的研究将允许建立消除内/溶酶体区室中病原体的新策略的概念证明。我们的研究成果将与野生、濒危和家养动物物种以及人类感染性疾病的治疗相关，并将开辟无数途径来检查抗病原体肽在病毒、细菌和原虫感染中的传递和作用。