Advancing the understanding of pathogenesis and virulence of Leptospira interrogans through proteomics, structural, mutagenesis and immunological analyses

• Grant number: 19/17488-2
• Support Opportunities: Research Projects - Thematic Grants
• Start date: December 01, 2020
• End date: November 30, 2025
• Field of knowledge: Biological Sciences - Microbiology - Applied Microbiology
• Principal Investigator: Ana Lucia Tabet Oller do Nascimento
• Grantee: Ana Lucia Tabet Oller do Nascimento
• Host Institution: Instituto Butantan. Secretaria da Saúde (São Paulo - Estado). São Paulo , SP, Brazil
• Pesquisadores principais: Daniel Carvalho Pimenta
• Associated researchers: Alexandre Paulo Yague Lopes ; Aline Florencio Teixeira ; Eliete Caló Romero ; Giovana Cappio Barazzone ; Gisele Oliveira de Souza ; Jademilson Celestino dos Santos ; Luis Guilherme Virgílio Fernandes ; Marcos Bryan Heinemann ; Mônica Larucci Vieira ; Silvio Arruda Vasconcellos
• Associated research grant(s): 25/08368-4 - ASTMH 2025 Annual Meeting, AR.EXT; 23/10870-4 - 58th Conference of the Brazilian Society of Tropical Medicine (MEDTROP 2023), AR.BR
• Associated scholarship(s): 23/08486-1 - "Structural, functional and immunogenic characterization of two proteins of Leptospira interrogans containing FecR domains", BP.MS; 22/16481-7 - Obtaining Leucine Rich Repeats (LRR) protein mutants in Leptospira interrogans and phenotypic evaluation of Host-pathogen interactions, BP.DD; 22/12101-5 - Expression of multifunctional protein Lsa23 (LIC11360) from Leptospira interrogans in the surrogate saprophyte L. biflexa and gain of function validation, BP.PD; + associated scholarships 22/08985-5 - Assessment of the role of two proteins of Leptospira interrogans in the pathogenesis and in the immunoprotective activity in animal model, BP.DD; 17/06731-8 - Application of CRISPR-interference for elucidating leptospirosis pathogenesis and development of novel strategies for knockout mutant's obtainment, BP.PD; 16/11541-0 - Evaluation of the vaccine potential of antigenic fractions obtained in Leptospira interrogans serovar Copenhageni using hamsters as an animal model, BP.PD - associated scholarships

Abstract

Leptospirosis is a zoonosis widely distributed in all continents, but prevalent in tropical and sub-tropical regions. The etiological agent is the pathogenic bacteria of the genus Leptospira. In developed countries, the disease in humans is associated with recreational activities with water, while in developing ones, the disease represents a public health problem, with 1 million cases reported being fatal cases around 60 thousands per year. In all countries, leptospirosis has an economic impact because it affects the livestock, causing loss of milk production, failure to thrive and premature death. In urban areas, the rodents, such as Rattus norvegicus, are the main reservoir of the disease. They are asymptomatic carriers of the bacteria, keeping shedding them live through the urine for their lifetime, contaminating soil and water. In Brazil, outbreaks of leptospirosis occur during the rainy season when flooding are frequent. Inundation in association with precarious sanitary conditions favors the contact of humans with contaminated water, spreading the disease. The symptoms of the disease are non-specific flu-like and include fever, headache and myalgia. In some cases, the disease evolves to severe forms, called Well's syndrome and Severe Pulmonary Hemorrhagic Syndrome- SPHS, with mortality rates in adults of 10 to 50% of the cases. There are no efficient diagnostic methods available. Because sanitary measures and rat eradication are difficult to implement, the adoption of prophylactic actions is the best choice to fight the disease. Therefore, different research groups around the world are pursuing the development of vaccine. Understanding the mechanisms of pathogenicity and the identification of virulent factors are crucial to block this pathogen. In this sense, the present project proposes distinctive methodological approaches aiming to advance the knowledge of these bacteria in different aspects: comparative proteomics analysis, structural biochemistry for the recombinant proteins available in our laboratory, mutants knock-in of saprophyte strain and gene silencing of pathogenic strain by CRISPR Cas9 method, scale-up of recombinant protein purification process for diagnostic kit development; to evaluate the involvement of the leptospiral proteins LipL41, LipL21, Lip46 and OmpL37, as potential receptor for endothelial and epithelial cultured cells in vitro. (AU)

Scientific publications

(13)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

TAKAHASHI, MARIA BEATRIZ; TEIXEIRA, ALINE FLORENCIO; NASCIMENTO, ANA LUCIA TABET OLLER. Overcoming problems to produce the recombinant protein LipL21 of Leptospira interrogans. BIOTECHNIQUES, v. N/A, p. 6-pg., 2023-04-17. (14/50981-0, 19/17488-2, 17/26223-7, 16/11541-0)

TAKAHASHI, M. B.; TEIXEIRA, A. F.; ALTO, NASCIMENTO. The leptospiral LipL21 and LipL41 proteins exhibit a broad spectrum of interactions with host cell components. VIRULENCE, v. 12, n. 1, p. 2798-2813, DEC 31 2021. (17/26223-7, 19/17488-2, 17/00236-5)

DAMIANO, DEBORAH K.; AZEVEDO, BRUNA O. P.; FERNANDES, GEORGE S. C.; TEIXEIRA, ALINE F.; GONCALVES, VIVIANE M.; NASCIMENTO, ANA L. T. O.; LOPES, ALEXANDRE P. Y.. The Toxin of VapBC-1 Toxin-Antitoxin Module from Leptospira interrogans Is a Ribonuclease That Does Not Arrest Bacterial Growth but Affects Cell Viability. MICROORGANISMS, v. 12, n. 8, p. 14-pg., 2024-08-01. (18/05823-9, 19/17488-2)

CAVENAGUE, MARIA F. F.; TEIXEIRA, ALINE F. F.; FERNANDES, LUIS G. V.; NASCIMENTO, ANA L. T. O.. LIC12254 Is a Leptospiral Protein That Interacts with Integrins via the RGD Motif. TROPICAL MEDICINE AND INFECTIOUS DISEASE, v. 8, n. 5, p. 15-pg., 2023-04-26. (18/08131-0, 19/17488-2, 16/11541-0, 17/06731-8)

SILVA, IGOR R. M.; TAKAHASHI, MARIA B.; TEIXEIRA, ALINE F.; NASCIMENTO, ANA L. T. O.. Evaluation of binding activities of a putative lipoprotein LIC_13355 of Leptospira spp.. FEBS OPEN BIO, v. N/A, p. 15-pg., 2024-12-12. (19/17488-2, 22/08985-5, 17/26223-7, 16/11541-0, 19/05466-4)

FERNANDES, LUIS G. V.; PUTZ, ELLIE J.; STASKO, JUDITH; LIPPOLIS, JOHN D.; NASCIMENTO, ANA L. T. O.; NALLY, JARLATH E.. valuation of LipL32 and LigA/LigB Knockdown Mutants in Leptospira interrogans Serovar Copenhageni: Impacts to Proteome and Virulenc. FRONTIERS IN MICROBIOLOGY, v. 12, FEB 2 2022. (17/06731-8, 19/20302-8, 19/17488-2)

PASSALIA, FELIPE JOSE; HEINEMANN, MARCOS BRYAN; VIEIRA, MONICA LARUCCI; NASCIMENTO, ANA LUCIA T. O.. A Novel Leptospira interrogans Protein LIC13086 Inhibits Fibrin Clot Formation and Interacts With Host Components. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, JUL 1 2021. (17/01102-2, 18/07054-2, 17/00236-5, 14/50981-0, 19/17488-2)

GASPAR, JOAO P.; TAKAHASHI, MARIA B.; TEIXEIRA, ALINE F.; NASCIMENTO, ANA L. T. O.. In silico analysis and functional characterization of a leucine-rich repeat protein of Leptospira interrogans. INTERNATIONAL JOURNAL OF MEDICAL MICROBIOLOGY, v. 316, p. 11-pg., 2024-09-03. (19/17488-2, 19/05466-4, 16/11541-0, 23/08486-1, 17/26223-7)

FERNANDES, LUIS GUILHERME V.; FOLTRAN, BRUNO B.; TEIXEIRA, ALINE F.; NASCIMENTO, ANA LUCIA TABET OLLER. LipL41 and LigA/LigB Gene Silencing on a LipL32 Knockout Leptospira interrogans Reveals the Impact of Multiple Mutations on Virulence. PATHOGENS, v. 12, n. 10, p. 12-pg., 2023-10-01. (19/17488-2, 17/06731-8, 16/11541-0, 22/16481-7)

FILHO, EDSON G. NASCIMENTO; VIEIRA, MONICA L.; DIAS, MERIELLEN; MENDES, MARIA A.; SANCHEZ, FABIO B.; SETUBAL, JOAO C.; HEINEMANN, MARCOS B.; SOUZA, GISELE O.; PIMENTA, DANIEL C.; NASCIMENTO, ANA L. T. O.. Global proteome of the saprophytic strain Leptospira biflexa and comparative analysis with pathogenic strain Leptospira interrogans uncover new pathogenesis mechanisms. JOURNAL OF PROTEOMICS, v. 297, p. 10-pg., 2024-02-20. (19/17488-2, 18/06201-1, 13/50218-2)

FERNANDES, LUIS G., V; AVELAR, KATIA E. S.; ROMERO, ELIETE C.; HEINEMANN, MARCOS B.; KIRCHGATTER, KARIN; NASCIMENTO, ANA L. T. O.. A New Recombinant Multiepitope Chimeric Protein of Leptospira interrogans Is a Promising Marker for the Serodiagnosis of Leptospirosis. TROPICAL MEDICINE AND INFECTIOUS DISEASE, v. 7, n. 11, p. 14-pg., 2022-11-01. (17/06731-8, 19/17488-2)

DAROZ, BRENDA B.; FERNANDES, LUIS G. V.; CAVENAGUE, MARIA F.; KOCHI, LEANDRO T.; PASSALIA, FELIPE J.; TAKAHASHI, MARIA B.; NASCIMENTO FILHO, EDSON G.; TEIXEIRA, ALINE F.; NASCIMENTO, ANA L. T. O.. A Review on Host-Leptospira Interactions: What We Know and Future Expectations. FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY, v. 11, p. 13-pg., 2021-11-25. (18/06201-1, 18/08131-0, 17/01102-2, 17/26223-7, 16/11541-0, 18/09652-4, 18/21959-8, 19/17488-2, 17/06731-8)

SANTOS, JADEMILSON C.; HANDA, SUMIT; FERNANDES, LUIS G. V.; BLEICHER, LUCAS; GANDIN, CESAR A.; DE OLIVEIRA-NETO, MARIO; GHOSH, PARTHO; NASCIMENTO, ANA LUCIA T. O.. Structural and biochemical characterization of Leptospira interrogans Lsa45 reveals a penicillin-binding protein with esterase activity. Process Biochemistry, v. 125, p. 13-pg., 2022-12-22. (14/50981-0, 17/25167-6, 18/20321-0, 19/17488-2, 17/06731-8)