Parasites of the trypanosomatid protozoan genus Leishmania are responsible for a spectrum of tropical diseases that afflict more than 10 million people worldwide, and depending on the specific species and immune status of the infected person, can be severe or fatal. In Brasil, Africa and the Mediterranean basin, leishmaniasis is a common opportunistic infection in AIDS patients, and many US soldiers now stationed in the Middle East have been infected. The goal of our research is to develop and apply new methods for the identification of genes used by this parasite to carry out its infectious cycle. Our premise is that the availability of powerful molecular genetic tools, and identification of genes important to parasite virulence, will radically advance our ability to develop improved control strategies. In previous studies powerful new methods were successfully developed for manipulating the parasite genome in a variety of ways, and modern 'genomic approaches'such as expression profiling and comparative genome hybridization applied. These studies were quite successful, but the number of exciting genes identified far exceeded the ability of our methods to tackle them. Fortunately we achieved two breakthroughs. First we showed that the RNA interference pathway was functional in species belong to the L. braziliensis species complex, despite its loss in other Leishmania species. We showed that the RNAi pathway could be used to conveniently and rapidly adapted to functionally assess gene function within the parasite. The features of this methodology promise to allow us to rapidly scale up and test many genes, and determine their role in virulence. The loss of this pathway in some Leishmania species is provocative and our studies suggest this is linked to important changes in the infectious cycle of great relevance to human disease. Our second breakthrough was the demonstration that it was possible to generate crosses of Leishmania experimentally. These new approaches will allow us to better describe the functional organization of the Leishmania genome, and by studying the inheritance of traits affecting animal virulence, to identify genes relevant to human disease in ways not previously possible.

Public Health Relevance

Leishmania are important tropical parasites, causing disease in more than 10 million people worldwide;more than 400 million people are at risk for infection in endemic regions. US military personnel have significant risk of infection in these areas as well, and recently concerns have risen that leishmaniasis may be endemic in certain dog populations in the USA. Depending on the species, Leishmania-induced pathology ranges from self-healing, cutaneous lesions to fatal, visceral diseases. Currently, there are no vaccines available against leishmaniasis, and the only approved chemotherapies are marginally effective, difficult to administer, and have significant associated toxicities. The underlying tenet of our research program is that improved understanding of key pathways required for parasite virulence and viability may provide opportunities for the development of improved therapies. In this project our goal is to develop and apply new methods for the identification of genes used by this parasite to carry out its infectious cycle. In previous studies powerful methods were successfully developed for manipulating the parasite genome in a variety of ways, and modern 'genomic approaches'such as expression profiling and comparative genome hybridization applied in several ways. These studies were quite successful, but the number of exciting genes identified far exceeded the ability of our methods to tackle them. Fortunately we achieved two breakthroughs. First we showed that the RNA interference pathway was functional in species belong to the L. braziliensis species complex, despite its loss in other Leishmania species. We showed that the RNAi pathway could be used to conveniently and rapidly adapted to functionally assess gene function within the parasite. The features of this methodology promise to allow us to rapidly scale up and test many genes, and determine their role in virulence. The loss of this pathway in some Leishmania species is provocative and our studies suggest this is linked to important changes in the infectious cycle of great relevance to human disease. Our second breakthrough was the demonstration that it was possible to generate crosses of Leishmania experimentally. These studies will allow us to better describe the functional organization of the Leishmania genome, and by studying the inheritance of traits affecting animal virulence in these studies, to identify genes relevant to human disease in ways not previously possible.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI029646-25
Application #
8602766
Study Section
Pathogenic Eukaryotes Study Section (PTHE)
Program Officer
Mcgugan, Glen C
Project Start
1990-03-01
Project End
2016-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
25
Fiscal Year
2014
Total Cost
Indirect Cost
Name
Washington University
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Mandell, Michael A; Beverley, Stephen M (2017) Continual renewal and replication of persistent Leishmania major parasites in concomitantly immune hosts. Proc Natl Acad Sci U S A 114:E801-E810
Kuhlmann, F Matthew; Robinson, John I; Bluemling, Gregory R et al. (2017) Antiviral screening identifies adenosine analogs targeting the endogenous dsRNA Leishmania RNA virus 1 (LRV1) pathogenicity factor. Proc Natl Acad Sci U S A 114:E811-E819
Matveyev, Andrey V; Alves, João M P; Serrano, Myrna G et al. (2017) The Evolutionary Loss of RNAi Key Determinants in Kinetoplastids as a Multiple Sporadic Phenomenon. J Mol Evol 84:104-115
Iantorno, Stefano A; Durrant, Caroline; Khan, Asis et al. (2017) Gene Expression in Leishmania Is Regulated Predominantly by Gene Dosage. MBio 8:
Rossi, Matteo; Castiglioni, Patrik; Hartley, Mary-Anne et al. (2017) Type I interferons induced by endogenous or exogenous viral infections promote metastasis and relapse of leishmaniasis. Proc Natl Acad Sci U S A 114:4987-4992
Mandell, Michael A; Beverley, Stephen M (2016) Concomitant Immunity Induced by Persistent Leishmania major Does Not Preclude Secondary Re-Infection: Implications for Genetic Exchange, Diversity and Vaccination. PLoS Negl Trop Dis 10:e0004811
Reynolds, David L; Hofmeister, Brigitte T; Cliffe, Laura et al. (2016) Base J represses genes at the end of polycistronic gene clusters in Leishmania major by promoting RNAP II termination. Mol Microbiol 101:559-74
Parmentier, Laurent; Cusini, Alexia; Müller, Norbert et al. (2016) Severe Cutaneous Leishmaniasis in a Human Immunodeficiency Virus Patient Coinfected with Leishmania braziliensis and Its Endosymbiotic Virus. Am J Trop Med Hyg 94:840-3
Macedo, Diego H; Menezes-Neto, Armando; Rugani, Jeronimo M et al. (2016) Low frequency of LRV1 in Leishmania braziliensis strains isolated from typical and atypical lesions in the State of Minas Gerais, Brazil. Mol Biochem Parasitol 210:50-54
Brettmann, Erin A; Shaik, Jahangheer S; Zangger, Haroun et al. (2016) Tilting the balance between RNA interference and replication eradicates Leishmania RNA virus 1 and mitigates the inflammatory response. Proc Natl Acad Sci U S A 113:11998-12005

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