Evaluation of Synergy Between Host and Pathogen-directed Therapies Against Intracellular Leishmania Donovani

Overview

Journal Int J Parasitol Drugs Drug Resist

Publisher Elsevier

Specialty Parasitology

Date 2019 Sep 8

PMID 31493763

Citations 9

Authors

M Shamim Hasan Zahid

Monica M Johnson

Robert J Tokarski 2nd

Abhay R Satoskar

James R Fuchs

Eric M Bachelder

Kristy M Ainslie

Affiliations

Soon will be listed here.

Abstract

Visceral leishmaniasis (VL) is associated with treatment complications due to the continued growth of resistant parasites toward currently available pathogen-directed therapeutics. To limit the emergence and combat resistant parasites there is a need to develop new anti-leishmanial drugs and alternative treatment approaches, such as host-directed therapeutics (HDTs). Discovery of new anti-leishmanial drugs including HDTs requires suitable in vitro assay systems. Herein, we modified and evaluated a series of resazurin assays against different life-stages of the VL causing parasite, Leishmania donovani to identify novel HDTs. We further analyzed the synergy of combinatorial interactions between traditionally used pathogen-directed drugs and HDTs for clearance of intracellular L. donovani. The inhibitory concentration at 50% (IC) of the five evaluated therapies [amphotericin B (AMB), miltefosine, paromomycin, DNER-4, and AR-12 (OSU-03012)] was determined against promastigotes, extracellular amastigotes, and intracellular amastigotes of L. donovani via a resazurin-based assay and compared to image-based microscopy. Using the resazurin-based assay, all evaluated therapies showed reproducible anti-leishmanial activity against the parasite's different life-stages. These results were consistent to the traditional image-based technique. The gold standard of therapy, AMB, showed the highest potency against intracellular L. donovani, and was further evaluated for combinatorial effects with the HDTs. Among the combinations analyzed, pathogen-directed AMB and host-directed AR-12 showed a synergistic reduction of intracellular L. donovani compared to individual treatments. The modified resazurin assay used in this study demonstrated a useful technique to measure new anti-leishmanial drugs against both intracellular and extracellular parasites. The synergistic interactions between pathogen-directed AMB and host-directed AR-12 showed a great promise to combat VL, with the potential to reduce the emergence of drug-resistant strains.

Citing Articles

The Potential Use of Peptides in the Fight against Chagas Disease and Leishmaniasis.

Berhe H, Kumar Cinthakunta Sridhar M, Zerihun M, Qvit N Pharmaceutics. 2024; 16(2).

PMID: 38399281 PMC: 10892537. DOI: 10.3390/pharmaceutics16020227.

Label-Free Mass Spectrometry Proteomics Reveals Different Pathways Modulated in THP-1 Cells Infected with Therapeutic Failure and Drug Resistance Clinical Isolates.

Tagliazucchi L, Perea-Martinez A, Fiorini G, Manzano J, Genovese F, Garcia-Hernandez R ACS Infect Dis. 2023; 9(3):470-485.

PMID: 36762976 PMC: 10012269. DOI: 10.1021/acsinfecdis.2c00457.

The Anti- and Anti- Action of Copper(II) and Silver(I) 1,10-Phenanthroline-5,6-dione Coordination Compounds.

Oliveira S, Santos V, Devereux M, McCann M, Santos A, Branquinha M Pathogens. 2023; 12(1).

PMID: 36678418 PMC: 9865435. DOI: 10.3390/pathogens12010070.

Synthesis and Anti-Leishmanial Properties of Quinolones Derived from Zanthosimuline.

Jezequel G, Cardoso L, Olivon F, Dennemont I, Apel C, Litaudon M Molecules. 2022; 27(22).

PMID: 36431992 PMC: 9693141. DOI: 10.3390/molecules27227892.

Challenges and Tools for In Vitro Exploratory Screening in the Drug Development Process: An Updated Review.

Cohen A, Azas N Pathogens. 2021; 10(12).

PMID: 34959563 PMC: 8703296. DOI: 10.3390/pathogens10121608.

References

Sarker S, Nahar L, Kumarasamy Y . Microtitre plate-based antibacterial assay incorporating resazurin as an indicator of cell growth, and its application in the in vitro antibacterial screening of phytochemicals. Methods. 2007; 42(4):321-4. PMC: 1895922. DOI: 10.1016/j.ymeth.2007.01.006. View

Ephros M, Waldman E, Zilberstein D . Pentostam induces resistance to antimony and the preservative chlorocresol in Leishmania donovani promastigotes and axenically grown amastigotes. Antimicrob Agents Chemother. 1997; 41(5):1064-8. PMC: 163851. DOI: 10.1128/AAC.41.5.1064. View

Mishra J, Singh S . Miltefosine resistance in Leishmania donovani involves suppression of oxidative stress-induced programmed cell death. Exp Parasitol. 2013; 135(2):397-406. DOI: 10.1016/j.exppara.2013.08.004. View

Murray H, Brooks E, DeVecchio J, Heinzel F . Immunoenhancement combined with amphotericin B as treatment for experimental visceral leishmaniasis. Antimicrob Agents Chemother. 2003; 47(8):2513-7. PMC: 166064. DOI: 10.1128/AAC.47.8.2513-2517.2003. View

Purkait B, Kumar A, Nandi N, Sardar A, Das S, Kumar S . Mechanism of amphotericin B resistance in clinical isolates of Leishmania donovani. Antimicrob Agents Chemother. 2011; 56(2):1031-41. PMC: 3264217. DOI: 10.1128/AAC.00030-11. View

McNicholl B, McGrath J, Quinn J . Development and application of a resazurin-based biomass activity test for activated sludge plant management. Water Res. 2006; 41(1):127-33. DOI: 10.1016/j.watres.2006.10.002. View

Perez-Victoria F, Sanchez-Canete M, Seifert K, Croft S, Sundar S, Castanys S . Mechanisms of experimental resistance of Leishmania to miltefosine: Implications for clinical use. Drug Resist Updat. 2006; 9(1-2):26-39. DOI: 10.1016/j.drup.2006.04.001. View

Pescher P, Blisnick T, Bastin P, Spath G . Quantitative proteome profiling informs on phenotypic traits that adapt Leishmania donovani for axenic and intracellular proliferation. Cell Microbiol. 2011; 13(7):978-91. DOI: 10.1111/j.1462-5822.2011.01593.x. View

Squires K, Rosenkaimer F, Sherwood J, Forni A, Were J, Murray H . Immunochemotherapy for visceral leishmaniasis: a controlled pilot trial of antimony versus antimony plus interferon-gamma. Am J Trop Med Hyg. 1993; 48(5):666-9. DOI: 10.4269/ajtmh.1993.48.666. View

10.

Kulshrestha A, Bhandari V, Mukhopadhyay R, Ramesh V, Sundar S, Maes L . Validation of a simple resazurin-based promastigote assay for the routine monitoring of miltefosine susceptibility in clinical isolates of Leishmania donovani. Parasitol Res. 2012; 112(2):825-8. DOI: 10.1007/s00436-012-3212-3. View

11.

Adler-Moore J, Proffitt R . AmBisome: liposomal formulation, structure, mechanism of action and pre-clinical experience. J Antimicrob Chemother. 2002; 49 Suppl 1:21-30. DOI: 10.1093/jac/49.suppl_1.21. View

12.

Zulfiqar B, Shelper T, Avery V . Leishmaniasis drug discovery: recent progress and challenges in assay development. Drug Discov Today. 2017; 22(10):1516-1531. DOI: 10.1016/j.drudis.2017.06.004. View

13.

Chiu H, Kulp S, Soni S, Wang D, Gunn J, Schlesinger L . Eradication of intracellular Salmonella enterica serovar Typhimurium with a small-molecule, host cell-directed agent. Antimicrob Agents Chemother. 2009; 53(12):5236-44. PMC: 2786354. DOI: 10.1128/AAC.00555-09. View

14.

Sundar S, Rai M, Chakravarty J, Agarwal D, Agrawal N, Vaillant M . New treatment approach in Indian visceral leishmaniasis: single-dose liposomal amphotericin B followed by short-course oral miltefosine. Clin Infect Dis. 2008; 47(8):1000-6. DOI: 10.1086/591972. View

15.

Miguel Neves B, Silvestre R, Resende M, Ouaissi A, Cunha J, Tavares J . Activation of phosphatidylinositol 3-kinase/Akt and impairment of nuclear factor-kappaB: molecular mechanisms behind the arrested maturation/activation state of Leishmania infantum-infected dendritic cells. Am J Pathol. 2010; 177(6):2898-911. PMC: 2993270. DOI: 10.2353/ajpath.2010.100367. View

16.

Mikus J, Steverding D . A simple colorimetric method to screen drug cytotoxicity against Leishmania using the dye Alamar Blue. Parasitol Int. 2001; 48(3):265-9. DOI: 10.1016/s1383-5769(99)00020-3. View

17.

Hudman D, Sargentini N . Resazurin-based assay for screening bacteria for radiation sensitivity. Springerplus. 2013; 2(1):55. PMC: 3586403. DOI: 10.1186/2193-1801-2-55. View

18.

Wadhone P, Maiti M, Agarwal R, Kamat V, Martin S, Saha B . Miltefosine promotes IFN-gamma-dominated anti-leishmanial immune response. J Immunol. 2009; 182(11):7146-54. DOI: 10.4049/jimmunol.0803859. View

19.

Musa A, Khalil E, Hailu A, Olobo J, Balasegaram M, Omollo R . Sodium stibogluconate (SSG) & paromomycin combination compared to SSG for visceral leishmaniasis in East Africa: a randomised controlled trial. PLoS Negl Trop Dis. 2012; 6(6):e1674. PMC: 3378617. DOI: 10.1371/journal.pntd.0001674. View

20.

Paape D, Bell A, Heal W, Hutton J, Leatherbarrow R, Tate E . Using a non-image-based medium-throughput assay for screening compounds targeting N-myristoylation in intracellular Leishmania amastigotes. PLoS Negl Trop Dis. 2014; 8(12):e3363. PMC: 4270692. DOI: 10.1371/journal.pntd.0003363. View