Transcriptome analysis uncovers the key pathways and candidate genes related to the treatment of Echinococcus granulosus protoscoleces with the repurposed drug pyronaridine

- Transcriptome analysis uncovers the key pathways and candidate genes related to the treatment of Echinococcus granulosus protoscoleces with the repurposed drug pyronaridine.
- Background: Cystic echinococcosis (CE) is a life-threatening zoonosis caused by the larval form of Echinococcus granulosus tapeworm.
- Interestingly, several heat shock protein (HSP) genes were greatly downregulated including stress-inducible HSPs and their constitutive cognates, and some of them belong to Echinococcus-specific expansion of HSP70..
- Keywords: Echinococcus granulosus, pyronaridine, RNA sequencing, Protoscoleces, MAPK, Heat shock protein, ABC transporter.
- 2021 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made.
- The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material.
- If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder.
- The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data..
- 1 NHC Key Laboratory of Parasite and Vector Biology, WHO Collaborating Centre for Tropical Diseases, National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, 200025 Shanghai, China Full list of author information is available at the end of the article.
- Cystic echinococcosis (CE) is a parasitic zoonosis caused by the larval stage of the dog tapeworm Echinococcus granulosus.
- granulosus leads to the development of one or multiple cysts located mostly in the liver and lungs, which triggers clinical signs in the late stage, including abdominal and chest pain, chronic cough, vomiting, even death [1, 2].
- Oral administration of PND showed high concentrations of the drug in the liver and lungs, which are the most affected organs in CE.
- However, the anti-CE mechanism of action of PND is not clear..
- Previous studies showed that the primary anti-malarial mode of action of PND is inhibition of β-hematin forma- tion, enhancement of hematin-induced red blood cells lysis, and inducing the formation of abnormal vesicles in the food vacuole of plasmodium [9, 10].
- In addition, PND has been char- acterized as a potential anticancer agent, which reverses the multi-drug‐resistance (MDR) phenotype in MDR cancer cell lines by inhibiting the function of the efflux P‐glycoprotein (Pgp) [12, 13].
- In this study, to obtain a comprehensive understanding of the anti-CE mechanism of PND, the global gene expression in E.
- Global gene expression of PND-treated E.
- The clean reads were mapped to the E.
- nlm.nih.gov/genome/?term=Echinococcus+granulosus).
- 1), a total of 1,321 genes were significantly differen- tially expressed in the PND-treated group compared to the control group, including 541 upregulated and 780 downregulated E.
- In addition, we found that PND treatment in- duced a significant downregulation in a large number of HSP genes including heat shock proteins (HSPs) and heat shock constitutive cognates (HSCs) (Table 4).
- Most of the genes are of HSP70 family and involved in the top two enriched pathways.
- To validate the results of transcriptome sequencing, quantitative PCR (qRT-PCR) was used for confirmation Table 1 Summary of read mapping results of the sequences generated from E.
- The qRT-PCR expression patterns of nine out of eleven DEGs were in agreement with the results of the transcriptome analysis, despite the variation of drug concentration or treatment time..
- 0.05), while in the case of EgMKK2, it was the treatment of LC 50 (p <.
- PND killed 100 % of the cysts in a mouse infection model by intraperitoneal injection at 57 mg/kg/day for three days.
- However, the anti-parasitic mechanism of PND remains unclear..
- Using a suitable low dose, our study showed that PND treatment induced changes in the expression of a large number of genes, including 541 E.
- The X-axis shows the differences in gene expression (FDR: adjusted p-value), while the Y-axis indicates expression changes (log2 fold change) of the genes in different groups.
- According to KEGG enrichment analysis, the MAPK pathway comes in second in the top-changed pathways affected by PND.
- In the last decade, a few components of the MAPK pathway have been identified in E.
- the key nodes of the MAPK pathway was not observed..
- In the genome of E.
- In this study, PND treatment Table 2 GO enrichment analysis of the DEGs of E.
- HSPs are originally identified because of their roles in response to heat shock (or other stressors) and these molecules are also molecular chaperones involved in protein folding and maturation [32].
- The differentially expressed HSP genes (Table 4) included five downregulated and three upregulated heat shock proteins and also eleven downregulated constitutive cog- nates, indicating that not all the DEGs observed in tran- scriptome analysis were necessarily induced by stress (e.g.
- In addition, most of the HSP genes are of the.
- The rich factor refers to the ratio of the DEGs annotated in the pathway versus the total number of genes annotated.
- Dot size is positively correlated to the number of DEGs.
- The colors of the dots represent the p values for the enrichment.
- Table 3 Differentially expressed ABC transporter genes following the PND treatment.
- HSPs are implicated in the cause and progression of various diseases, such as infections [35], cancer [36, 37], and neurodegeneration [38, 39].
- Some of the HSP genes have already been identi- fied and studied in E.
- Table 4 Differentially expressed HSP genes following the PND treatment Gene ID Log2 fold change (PND/.
- -3.81 Heat shock cognate.
- -3.29 Heat shock cognate.
- -3.77 Heat shock cognate.
- -4.96 Heat shock protein 70.
- -3.06 Heat shock cognate.
- -4.73 Heat shock cognate.
- -2.8 Heat shock cognate.
- -3.88 Heat shock cognate.
- 2.45 Heat shock protein.
- -5.86 Heat shock protein 70.
- 1.75 Small heat shock protein p36.
- -1.47 Heat shock cognate.
- -1.45 Heat shock protein.
- -2.31 Heat shock cognate.
- -2.71 Heat shock protein beta- 11.
- -2.1 Heat shock cognate.
- 1.93 Heat shock protein 70.
- -1.33 Heat shock cognate.
- -1.03 Heat shock 10 kda protein 1.
- Drug treatment of E.
- RNA-seq bioinformatics analysis.
- The high-quality clean reads were aligned to the reference genome using the HIASAT (https://ccb.jhu.edu/software/hisat2/index..
- nine downregulated genes: HSP72, ECSIT, SRF, SYF, LSM4, U2AF, and ABCB1) were selected from the top three enriched path- ways, and their gene expression levels in the control and PND-treated groups were evaluated.
- 3 Verification of the RNA-seq data by qRT-PCR.
- Bars represent the mean fold changes in the expression of eleven genes in E.
- PSCs used for RNA- seq were treated with PND at the concentration of LC 50 (49.0 µM) for 24 h.
- CE: Cystic echinococcosis.
- HSPs: Heat shock proteins.
- MAPK: Mitogen-activated protein kinase.
- RNA-seq: RNA sequencing.
- We appreciate the generous gift of the EgMKK1/EgMKK2 anti-serums pro- vided by Dr.
- WZ and LPD reviewed and revised the drafts of the manuscript.
- The RNA-seq data obtained in this study were deposited in the National Center for Biotechnology Information (NCBI) Sequence Read Archive (SRA) database (https://www.ncbi.nlm.nih.gov/sra) under accession number of PRJNA667188..
- 4 Western blot analysis of the effect of PND on EgMKK1 and EgMKK2.
- Cystic echinococcosis.
- Advances in the 21st Century.
- Old drug repurposing for neglected disease: Pyronaridine as a promising candidate for the treatment of Echinococcus granulosus infections.
- Global gene expression profiling of Plasmodium falciparum in response to the anti- malarial drug pyronaridine.
- The genome of the hydatid tapeworm Echinococcus granulosus.
- In vitro and in vivo efficacies of novel carbazole aminoalcohols in the treatment of cystic echinococcosis.
- Identification of functional MKK3/6 and MEK1/2 homologs from Echinococcus granulosus and investigation of protoscolecidal activity of mitogen-activated protein kinase signaling pathway inhibitors in vitro and in vivo.
- Molecular characterization of a signal-regulated kinase homolog from Echinococcus granulosus.
- Molecular cloning and characterization of a p38-like mitogen-activated protein kinase from Echinococcus granulosus.
- In vitro effects of SB202190 on Echinococcus granulosus.
- P-glycoprotein expression and pharmacological modulation in larval stages of Echinococcus granulosus.
- Increased expression of ABC transport proteins is associated with ivermectin resistance in the model nematode Caenorhabditis elegans.
- Expression of ATP-binding cassette multidrug transporters in the giant liver fluke Fasciola gigantica and their possible involvement in the transport of bile salts and anthelmintics.
- Heat shock protein (HSP) drug discovery and development: Targeting heat shock proteins in disease.
- Characterization of a putative hsp70 pseudogene transcribed in protoscoleces and adult worms of Echinococcus granulosus.
- Heat shock protein 90 inhibitors as broad spectrum anti-infectives.
- Association of heat-shock proteins in various neurodegenerative disorders: is it a master key to open the therapeutic door? Mol Cell Biochem.
- Heat shock proteins in neurodegenerative diseases: pathogenic roles and therapeutic implications.
- The Plasmodium falciparum Hsp70- x chaperone assists the heat stress response of the malaria parasite.
- A chemical inhibitor of heat shock protein 78 (HSP78) from Leishmania donovani represents a potential antileishmanial drug candidate.
- Molecular and biochemical mining of heat-shock and 14-3-3 proteins in drug-induced protoscolices of Echinococcus granulosus and the detection of a candidate gene for anthelmintic resistance.
- Molecular and immunological characterization of the C-terminal region of a new Echinococcus granulosus Heat Shock Protein 70.

Xem thử không khả dụng, vui lòng xem tại trang nguồn
hoặc xem Tóm tắt