Amphotericin B-Loaded Extracellular Vesicles Derived from Leishmania major Enhancing Cutaneous Leishmaniasis Treatment through In Vitro and In Vivo Studies
-
Afshin Davari
,
Homa Hajjaran
,
Ali Khamesipour
,
Mehdi Mohebali
,
Fatemeh Mehryab
,
Saeed Shahsavari
,
Faezeh Shekari
Abstract
Background: Recent studies have shown an increasing number of patients with cutaneous leishmaniasis (CL) who do not respond to pentavalent antimonials as the first line of treatment for CL. Nanocarriers such as extracellular vesicles (EVs) are efficient vehicles that might be used as drug delivery systems for the treatment of diseases. Therefore, we aimed to isolate and characterize the EVs of Leishmania major, load them with Amphotericin B (AmB), and investigate the toxicity and efficacy of the prepared drug form.
Methods: The EVs of L. major were isolated, characterized, and loaded with amphotericin B (AmB), and the EVs-Amphotericin B (EVs-AmB) form was synthesized. Relevant in vitro and in vivo methods were performed to evaluate the toxicity and efficacy of EVs-AmB compared to the control.
Results: The anti-leishmanial activity of the EVs-AmB showed a higher percentage inhibition (PI%) (P = 0.023) compared to the AmB at different concentrations and time points. Obtained data showed a significant increase in the lesion size and parasite load in the lesion, PBS, and EVs mice groups in comparison with EVs-AmB, AmB, and Glucantime groups (P < 0.05), EVs-AmB had a significant decrease in lesion sizes in comparison with AmB (P < 0.05). Results showed that EVs-AmB decreased its toxicity to the kidneys and liver (P < 0.05).
Conclusion: EVs-AmB improved the efficacy of AmB in mouse skin lesions and reduced hepatorenal toxicity. Furthermore, EVs could be a promising nanoplatform for the delivery of AmB in CL caused by L. major.
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2. Alvar J, Vélez ID, Bern C, et al. Leishmaniasis worldwide and global estimates of its incidence. PloS One. 2012;7(5):e35671.
3. Aronson N, Herwaldt BL, Libman M, et al. Diagnosis and treatment of leishmaniasis: clinical practice guidelines by the Infectious Diseases Society of America (IDSA) and the American Society of Tropical Medicine and Hygiene (ASTMH). Clin Infect Dis. 2016;63(12):e202-e64.
4. Nemati S, Fazaeli A, Hajjaran H, et al. Genetic diversity and phylogenetic analysis of the Iranian Leishmania parasites based on HSP70 gene PCR-RFLP and sequence analysis. Korean J Parasitol. 2017; 55(4):367-374.
5. Rostamian M, Bashiri H, Yousefinejad V, et al. Prevalence of human visceral leishmaniasis in Iran: A systematic review and meta-analysis. Comp Immunol Microbiol Infect Dis. 2021; 75:101604.
6. Madusanka RK, Silva H, Karunaweera ND. Treatment of cutaneous leishmaniasis and insights into species-specific responses: a narrative review. Infect Dis Therapy. 2022;11(2):695-711.
7. WHO. Leishmaniasis, Neglected tropical diseases. 2022. https://www.who.int/data/gho/data/themes/topics/gho-ntd-leishmaniasis. (accessed 2021).
8. Hajjaran H, Saberi R, Borjian A, et al. The geographical distribution of human cutaneous and visceral Leishmania species identified by molecular methods in Iran: a systematic review with meta-analysis. Front Public Health. 2021;9: 661674.
9. Razavi MR, Shirzadi MR, Mohebali M, et al. Human Cutaneous Leishmaniosis in Iran, Up to Date-2019. J Arthropod Borne Dis. 2021;15(2):143-151.
10. Sabzevari S, Teshnizi SH, Shokri A, et al. Cutaneous leishmaniasis in Iran: A systematic review and meta-analysis. Microb Pathog. 2021;152:104721.
11. López L, Robayo M, Vargas M. Thermotherapy. An alternative for the treatment of American cutaneous leishmaniasis. Trials. 2012;13:58.
12. Mohammad N, Sagheer F, Paracha MM, et al. Comparison Of Efficacy Of Miltefosine Versus Meglumine Antimonate In The Treatment Of Cutaneous Leishmaniasis. J Ayub Med College, JAMC. 2022;34(4):849-53.
13. Ponte-Sucre A, Gamarro F, Dujardin J-C, et al. Drug resistance and treatment failure in leishmaniasis: A 21st century challenge. PLoS Negl Trop Dis. 2017;11(12):e0006052.
14. Eskandari SE, Khamesipour A, Jaafari MR, et al. Combination of topical liposomal amphotericin B and Glucantime in comparison with glucantime alone for the treatment of anthroponotic cutaneous leishmaniasis (ACL). Iranian J Microb. 2021;13(5):718-23.
15. Mohtasebi S, Mohebali M, Elikaee S, et al. In vitro and in vivo anti-parasitic activity of biogenic antimony sulfide nanoparticles on Leishmania major (MRHO/IR/75/ER). Parasitol Res. 2019;118(9):2669-78.
16. Bayat F, Hosseinpour R, Mehryab F, et al. Potential application of liposomal nanodevices for non-cancer diseases: an update on design, characterization and biopharmaceutical evaluation. Adv Colloid Interface Sci. 2020;277:102121.
17. Ghorbani M, Farhoudi R. Leishmaniasis in humans: drug or vaccine therapy? Drug Des Devel Ther. 2018; 12:25-40.
18. Akbari M, Oryan A, Hatam G. Application of nanotechnology in treatment of leishmaniasis: a review. Acta Trop. 2017;172:86-90.
19. Coakley G, Maizels RM, Buck AH. Exosomes and other extracellular vesicles: the new communicators in parasite infections. Trend Parasitol. 2015;31(10):477-89.
20. Dong G, Filho AL, Olivier M. Modulation of host-pathogen communication by extracellular vesicles (EVs) of the protozoan parasite Leishmania. Front Cell Infect Microb. 2019;9:100.
21. Mehryab F, Rabbani S, Shahhosseini S, et al. Exosomes as a next-generation drug delivery system: an update on drug loading approaches, characterization, and clinical application challenges. Acta Biomater. 2020;113:42-62.
22. Soleymani M, Saberi S, Shekari F. Extracellular vesicles in regenerative medicine, a brief review. Modern Med Lab J. 2019;2(2):118-26.
23. Rai A, Fang H, Fatmous M, et al. A protocol for isolation, purification, characterization, and functional dissection of exosomes. Methods Mol Biol. 2021:2261:105-149.
24. Brennan K, Martin K, FitzGerald S.P, et al. A comparison of methods for the isolation and separation of extracellular vesicles from protein and lipid particles in human serum. Sci Rep. 2020;10(1):1039.
25. Heidari S, Hajjaran H, Kazemi B, et al. Identification of immunodominant proteins of Leishmania infantum by immunoproteomics to evaluate a recombinant multi-epitope designed antigen for serodiagnosis of human visceral leishmaniasis. Exp Parasitol. 2021;222:108065.
26. Mardpour S, Ghanian MH, Sadeghi-Abandansari H, et al. Hydrogel-mediated sustained systemic delivery of mesenchymal stem cell-derived extracellular vesicles improves hepatic regeneration in chronic liver failure. ACS Appl Mater Interfaces. 2019;11(41):37421-33.
27. Seyedrazizadeh S-Z, Poosti S, Nazari A, et al. Extracellular vesicles derived from human ES-MSCs protect retinal ganglion cells and preserve retinal function in a rodent model of optic nerve injury. Stem Cell Res Ther. 2020;11(1):203.
28. Parvez S, Yadagiri G, Gedda MR, et al. Modified solid lipid nanoparticles encapsulated with Amphotericin B and Paromomycin: an effective oral combination against experimental murine visceral leishmaniasis. Sci Rep. 2020;10(1): 12243.
29. Abu Ammar A, Nasereddin A, Ereqat S, et al. Amphotericin B-loaded nanoparticles for local treatment of cutaneous leishmaniasis. Drug Deli Trans Res. 2019;9(1):76-84.
30. Salehi-Sangani G, Mohebali M, Jajarmi V, et al. Immunization against Leishmania major infection in BALB/c mice using a subunit-based DNA vaccine derived from TSA, LmSTI1, KMP11, and LACK predominant antigens. Iran J Basic Med Sci. 2019;22(12):1493-1501.
31. Mehrizi TZ, Ardestani MS, Molla Hoseini MH, et al. Novel nano-sized chitosan amphotericin B formulation with considerable improvement against Leishmania major. Nanomedicine. 2018;13(24):3129-47.
32. Shirzadi MR. Lipsosomal amphotericin B: a review of its properties, function, and use for treatment of cutaneous leishmaniasis. Res Rep Trop Med. 2019; 10:11-18.
33. WHO. Regional Office for the Eastern Mediterranean. Manual for case management of cutaneous leishmaniasis in the WHO Eastern Mediterranean Region. https://apps.who.int/iris/handle/10665/120002. (accessed 2014).
34. Asad M, Bhattacharya P, Banerjee A. Therapeutic and immunomodulatory activities of short-course treatment of murine visceral leishmaniasis with KALSOME™ 10, a new liposomal amphotericin B. BMC Infect Dis. 2015;15:188.
35. Sundar S, Chakravarty J, Meena LP. Leishmaniasis: treatment, drug resistance and emerging therapies. Expert Opin Orphan Drug. 2019;7(1):1-10.
36. Bunggulawa EJ, Wang W, Yin T, et al. Recent advancements in the use of exosomes as drug delivery systems. J Nanobiotechnology. 2018;16(1):18.
37. Carrera-Bravo C, Koh EY, Tan KS. The roles of parasite-derived extracellular vesicles in disease and host-parasite communication. Parasitol Int. 2021;83:102373.
38. Pérez-Cabezas B, Santarém N, Cecílio P, et al. More than just exosomes: distinct Leishmania infantum extracellular products potentiate the establishment of infection. J Extracell Vesicles. 2018;7(1):1541708.
39. Kanchanapally R, Deshmukh SK, Chavva SR, et al. Drug-loaded exosomal preparations from different cell types exhibit distinctive loading capability, yield, and antitumor efficacies: a comparative analysis. Int J Nanomedicine. 2019;11:531-41.
40. Ebrahimian M, Hashemi M, Etemad L, et al. Thymoquinone-loaded mesenchymal stem cell-derived exosome as an efficient nano-system against breast cancer cells. Iran J Basic Med Sci. 2022;25(6):723-31.
41. Mishra J, Dey A, Singh N, Somvanshi R, et al. Evaluation of toxicity & therapeutic efficacy of a new liposomal formulation of amphotericin B in a mouse model. Indian J Med Res. 2013;137(4):767-76.
2. Alvar J, Vélez ID, Bern C, et al. Leishmaniasis worldwide and global estimates of its incidence. PloS One. 2012;7(5):e35671.
3. Aronson N, Herwaldt BL, Libman M, et al. Diagnosis and treatment of leishmaniasis: clinical practice guidelines by the Infectious Diseases Society of America (IDSA) and the American Society of Tropical Medicine and Hygiene (ASTMH). Clin Infect Dis. 2016;63(12):e202-e64.
4. Nemati S, Fazaeli A, Hajjaran H, et al. Genetic diversity and phylogenetic analysis of the Iranian Leishmania parasites based on HSP70 gene PCR-RFLP and sequence analysis. Korean J Parasitol. 2017; 55(4):367-374.
5. Rostamian M, Bashiri H, Yousefinejad V, et al. Prevalence of human visceral leishmaniasis in Iran: A systematic review and meta-analysis. Comp Immunol Microbiol Infect Dis. 2021; 75:101604.
6. Madusanka RK, Silva H, Karunaweera ND. Treatment of cutaneous leishmaniasis and insights into species-specific responses: a narrative review. Infect Dis Therapy. 2022;11(2):695-711.
7. WHO. Leishmaniasis, Neglected tropical diseases. 2022. https://www.who.int/data/gho/data/themes/topics/gho-ntd-leishmaniasis. (accessed 2021).
8. Hajjaran H, Saberi R, Borjian A, et al. The geographical distribution of human cutaneous and visceral Leishmania species identified by molecular methods in Iran: a systematic review with meta-analysis. Front Public Health. 2021;9: 661674.
9. Razavi MR, Shirzadi MR, Mohebali M, et al. Human Cutaneous Leishmaniosis in Iran, Up to Date-2019. J Arthropod Borne Dis. 2021;15(2):143-151.
10. Sabzevari S, Teshnizi SH, Shokri A, et al. Cutaneous leishmaniasis in Iran: A systematic review and meta-analysis. Microb Pathog. 2021;152:104721.
11. López L, Robayo M, Vargas M. Thermotherapy. An alternative for the treatment of American cutaneous leishmaniasis. Trials. 2012;13:58.
12. Mohammad N, Sagheer F, Paracha MM, et al. Comparison Of Efficacy Of Miltefosine Versus Meglumine Antimonate In The Treatment Of Cutaneous Leishmaniasis. J Ayub Med College, JAMC. 2022;34(4):849-53.
13. Ponte-Sucre A, Gamarro F, Dujardin J-C, et al. Drug resistance and treatment failure in leishmaniasis: A 21st century challenge. PLoS Negl Trop Dis. 2017;11(12):e0006052.
14. Eskandari SE, Khamesipour A, Jaafari MR, et al. Combination of topical liposomal amphotericin B and Glucantime in comparison with glucantime alone for the treatment of anthroponotic cutaneous leishmaniasis (ACL). Iranian J Microb. 2021;13(5):718-23.
15. Mohtasebi S, Mohebali M, Elikaee S, et al. In vitro and in vivo anti-parasitic activity of biogenic antimony sulfide nanoparticles on Leishmania major (MRHO/IR/75/ER). Parasitol Res. 2019;118(9):2669-78.
16. Bayat F, Hosseinpour R, Mehryab F, et al. Potential application of liposomal nanodevices for non-cancer diseases: an update on design, characterization and biopharmaceutical evaluation. Adv Colloid Interface Sci. 2020;277:102121.
17. Ghorbani M, Farhoudi R. Leishmaniasis in humans: drug or vaccine therapy? Drug Des Devel Ther. 2018; 12:25-40.
18. Akbari M, Oryan A, Hatam G. Application of nanotechnology in treatment of leishmaniasis: a review. Acta Trop. 2017;172:86-90.
19. Coakley G, Maizels RM, Buck AH. Exosomes and other extracellular vesicles: the new communicators in parasite infections. Trend Parasitol. 2015;31(10):477-89.
20. Dong G, Filho AL, Olivier M. Modulation of host-pathogen communication by extracellular vesicles (EVs) of the protozoan parasite Leishmania. Front Cell Infect Microb. 2019;9:100.
21. Mehryab F, Rabbani S, Shahhosseini S, et al. Exosomes as a next-generation drug delivery system: an update on drug loading approaches, characterization, and clinical application challenges. Acta Biomater. 2020;113:42-62.
22. Soleymani M, Saberi S, Shekari F. Extracellular vesicles in regenerative medicine, a brief review. Modern Med Lab J. 2019;2(2):118-26.
23. Rai A, Fang H, Fatmous M, et al. A protocol for isolation, purification, characterization, and functional dissection of exosomes. Methods Mol Biol. 2021:2261:105-149.
24. Brennan K, Martin K, FitzGerald S.P, et al. A comparison of methods for the isolation and separation of extracellular vesicles from protein and lipid particles in human serum. Sci Rep. 2020;10(1):1039.
25. Heidari S, Hajjaran H, Kazemi B, et al. Identification of immunodominant proteins of Leishmania infantum by immunoproteomics to evaluate a recombinant multi-epitope designed antigen for serodiagnosis of human visceral leishmaniasis. Exp Parasitol. 2021;222:108065.
26. Mardpour S, Ghanian MH, Sadeghi-Abandansari H, et al. Hydrogel-mediated sustained systemic delivery of mesenchymal stem cell-derived extracellular vesicles improves hepatic regeneration in chronic liver failure. ACS Appl Mater Interfaces. 2019;11(41):37421-33.
27. Seyedrazizadeh S-Z, Poosti S, Nazari A, et al. Extracellular vesicles derived from human ES-MSCs protect retinal ganglion cells and preserve retinal function in a rodent model of optic nerve injury. Stem Cell Res Ther. 2020;11(1):203.
28. Parvez S, Yadagiri G, Gedda MR, et al. Modified solid lipid nanoparticles encapsulated with Amphotericin B and Paromomycin: an effective oral combination against experimental murine visceral leishmaniasis. Sci Rep. 2020;10(1): 12243.
29. Abu Ammar A, Nasereddin A, Ereqat S, et al. Amphotericin B-loaded nanoparticles for local treatment of cutaneous leishmaniasis. Drug Deli Trans Res. 2019;9(1):76-84.
30. Salehi-Sangani G, Mohebali M, Jajarmi V, et al. Immunization against Leishmania major infection in BALB/c mice using a subunit-based DNA vaccine derived from TSA, LmSTI1, KMP11, and LACK predominant antigens. Iran J Basic Med Sci. 2019;22(12):1493-1501.
31. Mehrizi TZ, Ardestani MS, Molla Hoseini MH, et al. Novel nano-sized chitosan amphotericin B formulation with considerable improvement against Leishmania major. Nanomedicine. 2018;13(24):3129-47.
32. Shirzadi MR. Lipsosomal amphotericin B: a review of its properties, function, and use for treatment of cutaneous leishmaniasis. Res Rep Trop Med. 2019; 10:11-18.
33. WHO. Regional Office for the Eastern Mediterranean. Manual for case management of cutaneous leishmaniasis in the WHO Eastern Mediterranean Region. https://apps.who.int/iris/handle/10665/120002. (accessed 2014).
34. Asad M, Bhattacharya P, Banerjee A. Therapeutic and immunomodulatory activities of short-course treatment of murine visceral leishmaniasis with KALSOME™ 10, a new liposomal amphotericin B. BMC Infect Dis. 2015;15:188.
35. Sundar S, Chakravarty J, Meena LP. Leishmaniasis: treatment, drug resistance and emerging therapies. Expert Opin Orphan Drug. 2019;7(1):1-10.
36. Bunggulawa EJ, Wang W, Yin T, et al. Recent advancements in the use of exosomes as drug delivery systems. J Nanobiotechnology. 2018;16(1):18.
37. Carrera-Bravo C, Koh EY, Tan KS. The roles of parasite-derived extracellular vesicles in disease and host-parasite communication. Parasitol Int. 2021;83:102373.
38. Pérez-Cabezas B, Santarém N, Cecílio P, et al. More than just exosomes: distinct Leishmania infantum extracellular products potentiate the establishment of infection. J Extracell Vesicles. 2018;7(1):1541708.
39. Kanchanapally R, Deshmukh SK, Chavva SR, et al. Drug-loaded exosomal preparations from different cell types exhibit distinctive loading capability, yield, and antitumor efficacies: a comparative analysis. Int J Nanomedicine. 2019;11:531-41.
40. Ebrahimian M, Hashemi M, Etemad L, et al. Thymoquinone-loaded mesenchymal stem cell-derived exosome as an efficient nano-system against breast cancer cells. Iran J Basic Med Sci. 2022;25(6):723-31.
41. Mishra J, Dey A, Singh N, Somvanshi R, et al. Evaluation of toxicity & therapeutic efficacy of a new liposomal formulation of amphotericin B in a mouse model. Indian J Med Res. 2013;137(4):767-76.
| Files | ||
| Issue | Vol 18 No 4 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v18i4.14260 | |
| Keywords | ||
| Leishmania major Extracellular vesicles Drug delivery Amphotericin B In vitro and In vivo. | ||
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How to Cite
1.
Davari A, Hajjaran H, Khamesipour A, Mohebali M, Mehryab F, Shahsavari S, Shekari F. Amphotericin B-Loaded Extracellular Vesicles Derived from Leishmania major Enhancing Cutaneous Leishmaniasis Treatment through In Vitro and In Vivo Studies. Iran J Parasitol. 2023;18(4):514-525.
