Periodic Reporting for period 1 - MetaLeish (Metals against Leishmaniasis)
Período documentado: 2023-04-01 hasta 2025-03-31
Leishmaniasis is one of the world’s neglected tropical diseases. It is caused by over 20 species of protozoan parasites of the Leishmania genus. Clinically, it manifests in three forms: cutaneous leishmaniasis
(CL), characterized by deep lacerations in the skin; mucocutaneus leishmaniasis (ML), characterized by partial or total destruction of mucous membranes of the nose, mouth and throat; and visceral leishmaniasis (VL), the deadliest, which affects the spleen, the liver and can reach the bone marrow.
The current treatments are few, in some cases expensive and with limited access. Currently available drugs were developed more than 50 years ago and not specifically for the treatment of leishmaniasis. Moreover, they can be
toxic to patients. Treatment involves the administration of pentavalent antimonial compounds (for instance, sodium stibogluconate or meglumine antimonite), or other drugs such as liposomal amphotericin B (L-AMB). The treatment time is long (up to 28 days) and the success rates varies widely due to either drop-out from the treatment, early death, migraines, anorexia, and others, which can be attributed to adverse effects from these drugs.
Within this scenario, the MetaLeish project (Metals against Leishmaniasis) aims at developing a new treatment based on biodegradable metal nanoparticles (BMPs) to cure cutaneous leishmaniasis. The purpose is to develop a new, practical, and affordable therapeutic solution that minimizes unwanted side effects, avoids the development of resistances, and is of easy access for the most affected population. The general hypothesis is that BMPs of metals such as zinc (Zn) or magnesium (Mg) can affect the intracellular environment where the parasite thrives and, thus, affect its viability.
(CL), characterized by deep lacerations in the skin; mucocutaneus leishmaniasis (ML), characterized by partial or total destruction of mucous membranes of the nose, mouth and throat; and visceral leishmaniasis (VL), the deadliest, which affects the spleen, the liver and can reach the bone marrow.
The current treatments are few, in some cases expensive and with limited access. Currently available drugs were developed more than 50 years ago and not specifically for the treatment of leishmaniasis. Moreover, they can be
toxic to patients. Treatment involves the administration of pentavalent antimonial compounds (for instance, sodium stibogluconate or meglumine antimonite), or other drugs such as liposomal amphotericin B (L-AMB). The treatment time is long (up to 28 days) and the success rates varies widely due to either drop-out from the treatment, early death, migraines, anorexia, and others, which can be attributed to adverse effects from these drugs.
Within this scenario, the MetaLeish project (Metals against Leishmaniasis) aims at developing a new treatment based on biodegradable metal nanoparticles (BMPs) to cure cutaneous leishmaniasis. The purpose is to develop a new, practical, and affordable therapeutic solution that minimizes unwanted side effects, avoids the development of resistances, and is of easy access for the most affected population. The general hypothesis is that BMPs of metals such as zinc (Zn) or magnesium (Mg) can affect the intracellular environment where the parasite thrives and, thus, affect its viability.
The main objectives/achiecements of this project were:
Manufacturing of MgO and ZnO nanoparticles through spark discharge generator (SDG), which was not achieved due to high contamination. Instead the particles were chemically synthesized.
Definition of a post-treatment protocol to ensure a stable colloidal suspension nanoparticles after fabrication, to maintain target dimensions in final liquid media to be administered in the cell culture. Particles were functionalized with two aminosilane groups: tetraethoxysilane (TEOS) and 3-aminopropyltriethoxysilane (APTES).
Determination of a non-lethal dose of Mg and Zn NPs, that are active against the amastigote parasites, effectively killing them, without affecting macrophage viability.
Description of the interaction of Mg and Zn NPs with macrophages (in terms metabolic processes, influence on the activation of macrophages into the M1 or M2 profile, production of ROS, among others) as well as their effect on Leishmania parasite.
Manufacturing of MgO and ZnO nanoparticles through spark discharge generator (SDG), which was not achieved due to high contamination. Instead the particles were chemically synthesized.
Definition of a post-treatment protocol to ensure a stable colloidal suspension nanoparticles after fabrication, to maintain target dimensions in final liquid media to be administered in the cell culture. Particles were functionalized with two aminosilane groups: tetraethoxysilane (TEOS) and 3-aminopropyltriethoxysilane (APTES).
Determination of a non-lethal dose of Mg and Zn NPs, that are active against the amastigote parasites, effectively killing them, without affecting macrophage viability.
Description of the interaction of Mg and Zn NPs with macrophages (in terms metabolic processes, influence on the activation of macrophages into the M1 or M2 profile, production of ROS, among others) as well as their effect on Leishmania parasite.
MgO and ZnO particles were successfully functionalized with two aminosilane groups: tetraethoxysilane (TEOS) and 3-aminopropyltriethoxysilane (APTES), which allowed higher colloidal stability in cell culture media.
Dose response curves of this for type of particles (functionalized and non-functionalized) were determined for THP-1 macrophages as well as two strains of associated with cutaneous leishmaniasis: L. braziliensis and L. major.
Non-toxic doses of each type of particles were tested against infected cells, for each parasites. MgO APTES particles accumulated in the cells and were stained, and thus their anti-leishmania activity could not be determined. All other particles shown anti-leishmania effect against one of both strains.
Molecular biology tests were conducted to determine particle interaction with any metabolic processes or influence on the activation of macrophages into the M1 or M2 profile. Nothing was detected at this point, which may indicate that the particles exert a direct action against the intracellular parasite in infected macrophages.
Dose response curves of this for type of particles (functionalized and non-functionalized) were determined for THP-1 macrophages as well as two strains of associated with cutaneous leishmaniasis: L. braziliensis and L. major.
Non-toxic doses of each type of particles were tested against infected cells, for each parasites. MgO APTES particles accumulated in the cells and were stained, and thus their anti-leishmania activity could not be determined. All other particles shown anti-leishmania effect against one of both strains.
Molecular biology tests were conducted to determine particle interaction with any metabolic processes or influence on the activation of macrophages into the M1 or M2 profile. Nothing was detected at this point, which may indicate that the particles exert a direct action against the intracellular parasite in infected macrophages.