Arthropod-borne Rickettsia species are among the most virulent and lethal human pathogens and are of significant global Public Health concern. Our work over the past 3.5 years resulted in significant progress in understanding of R. typhi pathogenesis, and in particular, the role of the rickettsial secretome during host cell invasion. For this proposal, we will address fundamental questions in two understudied yet highly significant topics in rickettsial pathogen biology. First, we will determine the mechanisms by which R. typhi modulates cat flea (Ctenocephalides felis) innate immunity, microbiota and other factors to establish infection and facilitate transmission to the vertebrate host; and second, defining the functional spectrum and immunogenic potential of the R. typhi armory of secreted phospholipases that alter host membrane physiology to facilitate phagosomal escape and intracellular replication. We will implement a transdisciplinary approach (e.g., phylogenomics, bacteriology, entomology, and cellular/molecular biology) to thoroughly investigate these focal areas, as outlined by the following Specific Aims.
In Aim I, we will decipher the modulatory factors R. typhi employs to colonize C. felis and facilitate transmission via 1) a robust tissue-specific, time-course expression analysis, 2) silencing IMD and Toll pathway gene expression, 3) testing the immunomodulatory propensities of isolated R. typhi peptidoglycan, and 4) characterizing the impact of the C. felis microbiome on infection.
In Aim II, we will characterize the function and immunogenicity of R. typhi secretory phospholipases. We will take two approaches to characterize R. typhi phospholipases: 1) functional characterization of Pat1, Pat2 and Pld via determining their subcellular localization, lipid recognition and interactome within host cells, with downstream analyses designed to reveal the significance of these interactions in mediating bacterial access to the host cytosol; and; 2) identification of inflammasome components necessary for IL-1? secretion by R. typhi, and determining the impact of Pat1, Pat2, and Pld activities on inflammasome activation. Together these studies will significantly advance our understanding of the complex Rickettsia/vector/host relationship, shedding light on how rickettsial parasites of arthropods transition to vertebrate pathogens throughout the obligatory Rickettsia life cycle. Thus, we anticipate that our work will take us closer to generating more prudent therapeutics to combat fatal rickettsioses.

Public Health Relevance

Rickettsia species, including agents of epidemic typhus, murine typhus and spotted fever, are among the most virulent human pathogens. Rickettsial diseases are seriously under-recognized and under-studied, and thus remain poorly appreciated as public health problems (particularly in the tropics). Using a multidisciplinary approach, here we will reveal functional characterization of several factors at the host/vector/pathogen interface, and anticipate that our work will take us closer to generating more prudent therapeutics to combat fatal rickettsioses.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI017828-36
Application #
9624376
Study Section
Vector Biology Study Section (VB)
Program Officer
Perdue, Samuel S
Project Start
1982-09-01
Project End
2023-05-31
Budget Start
2019-06-01
Budget End
2020-05-31
Support Year
36
Fiscal Year
2019
Total Cost
Indirect Cost
Name
University of Maryland Baltimore
Department
Microbiology/Immun/Virology
Type
Schools of Medicine
DUNS #
188435911
City
Baltimore
State
MD
Country
United States
Zip Code
21201
Gillespie, Joseph J; Driscoll, Timothy P; Verhoeve, Victoria I et al. (2018) A Tangled Web: Origins of Reproductive Parasitism. Genome Biol Evol 10:2292-2309
Lehman, Stephanie S; Noriea, Nicholas F; Aistleitner, Karin et al. (2018) The Rickettsial Ankyrin Repeat Protein 2 Is a Type IV Secreted Effector That Associates with the Endoplasmic Reticulum. MBio 9:
Rennoll, Sherri A; Rennoll-Bankert, Kristen E; Guillotte, Mark L et al. (2018) The Cat Flea (Ctenocephalides felis) Immune Deficiency Signaling Pathway Regulates Rickettsia typhi Infection. Infect Immun 86:
Hagen, Rachael; Verhoeve, Victoria I; Gillespie, Joseph J et al. (2018) Conjugative Transposons and Their Cargo Genes Vary across Natural Populations of Rickettsia buchneri Infecting the Tick Ixodes scapularis. Genome Biol Evol 10:3218-3229
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Driscoll, Timothy P; Verhoeve, Victoria I; Guillotte, Mark L et al. (2017) Wholly Rickettsia! Reconstructed Metabolic Profile of the Quintessential Bacterial Parasite of Eukaryotic Cells. MBio 8:
Gillespie, Joseph J; Phan, Isabelle Q H; Driscoll, Timothy P et al. (2016) The Rickettsia type IV secretion system: unrealized complexity mired by gene family expansion. Pathog Dis 74:
Rennoll-Bankert, Kristen E; Rahman, M Sayeedur; Guillotte, Mark L et al. (2016) RalF-Mediated Activation of Arf6 Controls Rickettsia typhi Invasion by Co-Opting Phosphoinositol Metabolism. Infect Immun 84:3496-3506
Gulia-Nuss, Monika; Nuss, Andrew B; Meyer, Jason M et al. (2016) Genomic insights into the Ixodes scapularis tick vector of Lyme disease. Nat Commun 7:10507
Gillespie, Joseph J; Kaur, Simran J; Rahman, M Sayeedur et al. (2015) Secretome of obligate intracellular Rickettsia. FEMS Microbiol Rev 39:47-80

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