Spider venom administration impairs glioblastoma growth and modulates immune response in a non-clinical model

Author(s):Show less -

Bonfanti, Amanda Pires ^{[1, 2]} ; Barreto, Natalia ^{[1, 2]} ; Munhoz, Jaqueline ^{[1, 2]} ; Caballero, Marcus ^{[1, 2]} ; Cordeiro, Gabriel ^{[1, 2]} ; Rocha-e-Silva, Thomaz ^[3] ; Sutti, Rafael ^[4] ; Moura, Fernanda ^[5] ; Brunetto, Sergio ^[6] ; Ramos, Celso Dario ^[7] ; Thome, Rodolfo ^[8] ; Verinaud, Liana ^[1] ; Raposo, Catarina ^[2]

Affiliation

^[1] Univ Estadual Campinas, Inst Biol, Dept Biol Estrutural & Func, Campinas, SP - Brazil

^[2] Univ Estadual Campinas UNICAMP, Fac Ciencias Farmaceut, Campinas, SP - Brazil

^[3] Fac Israelita Ciencias Saude Albert Einstein, Sao Paulo, SP - Brazil

^[4] Santa Casa Sao Paulo, Fac Ciencias Med, Sao Paulo, SP - Brazil

^[5] Univ Estadual Campinas, Fac Ciencias Med, Campinas, SP - Brazil

^[6] Univ Estadual Campinas, Ctr Engn Biomed, Campinas, SP - Brazil

^[7] Univ Estadual Campinas, Hosp Clin, Serv Med Nucl, Campinas, SP - Brazil

^[8] Thomas Jefferson Univ, Dept Neurol, Philadelphia, PA 19107 - USA

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 10, n. 1 APR 3 2020.
• Web of Science Citations: 1

Abstract

Molecules from animal venoms are promising candidates for the development of new drugs. Previous in vitro studies have shown that the venom of the spider Phoneutria nigriventer (PnV) is a potential source of antineoplastic components with activity in glioblastoma (GB) cell lines. In the present work, the effects of PnV on tumor development were established in vivo using a xenogeneic model. Human GB (NG97, the most responsive line in the previous study) cells were inoculated (s.c.) on the back of RAG(-/-) mice. PnV (100 mu g/Kg) was administrated every 48 h (i.p.) for 14 days and several endpoints were evaluated: tumor growth and metabolism (by microPET/CT, using F-18-FDG), tumor weight and volume, histopathology, blood analysis, percentage and profile of macrophages, neutrophils and NK cells isolated from the spleen (by flow cytometry) and the presence of macrophages (Iba-1 positive) within/surrounding the tumor. The effect of venom was also evaluated on macrophages in vitro. Tumors from PnV-treated animals were smaller and did not uptake detectable amounts of F-18-FDG, compared to control (untreated). PnV-tumor was necrotic, lacking the histopathological characteristics typical of GB. Since in classic chemotherapies it is observed a decrease in immune response, methotrexate (MTX) was used only to compare the PnV effects on innate immune cells with a highly immunosuppressive antineoplastic drug. The venom increased monocytes, neutrophils and NK cells, and this effect was the opposite of that observed in the animals treated with MTX. PnV increased the number of macrophages in the tumor, while did not increase in the spleen, suggesting that PnV-activated macrophages were led preferentially to the tumor. Macrophages were activated in vitro by the venom, becoming more phagocytic; these results confirm that this cell is a target of PnV components. Spleen and in vitro PnV-activated macrophages were different of M1, since they did not produce pro- and anti-inflammatory cytokines. Studies in progress are selecting the venom molecules with antitumor and immunomodulatory effects and trying to better understand their mechanisms. The identification, optimization and synthesis of antineoplastic drugs from PnV molecules may lead to a new multitarget chemotherapy. Glioblastoma is associated with high morbidity and mortality; therefore, research to develop new treatments has great social relevance. Natural products and their derivatives represent over one-third of all new molecular entities approved by FDA. However, arthropod venoms are underexploited, although they are a rich source of new molecules. A recent in vitro screening of the Phoneutria nigriventer spider venom (PnV) antitumor effects by our group has shown that the venom significantly affected glioblastoma cell lines. Therefore, it would be relevant to establish the effects of PnV on tumor development in vivo, considering the complex neoplastic microenvironment. The venom was effective at impairing tumor development in murine xenogeneic model, activating the innate immune response and increasing tumor infiltrating macrophages. In addition, PnV activated macrophages in vitro for a different profile of M1. These activated PnV-macrophages have potential to fight the tumor without promoting tumorigenesis. Studies in progress are selecting the venom molecules with antitumor and immunomodulatory effects and trying to better understand their mechanisms. We aim to synthesize and carry out a formulation with these antineoplastic molecules for clinical trials. Spider venom biomolecules induced smaller and necrotic xenogeneic GB; spider venom activated the innate immune system; venom increased blood monocytes and the migration of macrophages to the tumor; activated PnV-macrophages have a profile different of M1 and have a potential to fight the tumor without promote tumorigenesis.Y (AU)

• FAPESP's process: 18/03051-9 - Identification of macrophages modulators molecules from spider venom: new perspectives for the treatment of cancer
• Grantee: Jaqueline de Lima Munhoz
• Support type: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 15/06134-4 - Multi-User Equipment approved in grant 2014/03002-7: cell imaging multi-mode reader
• Grantee: Marcelo Bispo de Jesus
• Support type: Multi-user Equipment Program
• FAPESP's process: 16/15827-6 - Identification of new molecules with chemotherapeutic effect in human glioma and characterization of the mechanism
• Grantee: Catarina Raposo Dias Carneiro
• Support type: Scholarships in Brazil - Young Researchers
• FAPESP's process: 17/16196-2 - Analysis of the effects and mechanisms of the Phoneutria nigriventer spider venom on morphology and migration of tumor cells
• Grantee: Natália Barreto dos Santos
• Support type: Scholarships in Brazil - Master
• FAPESP's process: 18/23559-7 - Cell therapy with macrophages and NK cells modulated ex vivo by peptide isolated from animal venom: a new approach in immunotherapy for cancer
• Grantee: Amanda Pires Bonfanti
• Support type: Scholarships in Brazil - Doctorate
• FAPESP's process: 17/05402-0 - Identification and characterization of spider Phoneutria nigriventer isolated venom, with chemotherapeutic effect on brain tumor cells
• Grantee: Amanda Pires Bonfanti
• Support type: Scholarships in Brazil - Technical Training Program - Technical Training
• FAPESP's process: 15/04194-0 - Identification of new molecules with chemotherapeutic effect in human glioma and characterization of the mechanism
• Grantee: Catarina Raposo Dias Carneiro
• Support type: Research Grants - Young Investigators Grants