Lyme disease (LD), caused by the spirochete Borrelia burgdorferi (Bb), is the most common arthropod-borne infection in the United States. Bb is maintained in nature via an enzootic cycle that typically involves rodents and Ixodes ticks. To sustain this complex life cycle, Bb must continuously alter its transcriptome, proteome, and metabolome in response to arthropod- and mammalian host-derived signals. Our long term objective has been to characterize the environmental cues, regulatory pathways, and genes critical for these adaptive changes. In our previously funded application, we hypothesized that the alternative sigma factor RpoS acts as a 'gatekeeper' that (i) transcribes genes required for tick transmission as well as the establishment of mammalian infection and (ii) represses tick-phase genes during mammalian host-adaptation. Our validation of this hypothesis has yielded powerful new insights into the mechanisms that perpetuate the bacterium in nature and maintain Lyme disease as a global threat to human health. A number of our recent findings relate to the transmission of spirochetes during nymphal feeding, the seminal event for infection of humans as well as mice. Using methodologies developed for tracking and imaging live spirochetes within ticks, we discovered that dissemination of Bb in feeding nymphs involves an initial non-motile phase (termed 'adherence-mediated migration') followed by a transition to motility that enables individual organisms to penetrate the midgut. These studies have helped elucidate the range of adaptations, physiological as well as structural, that spirochetes must undergo in order to negotiate the formidable barriers they encounter en route to the mammal. They also have definitively demonstrated that RpoS-regulated genes are required for transmission of Bb (see Progress Report). Our collective results give rise to our primary hypothesis, to be assessed in Aims 1 and 2, that the RpoS regulon contains many uncharacterized genes that promote the biphasic dissemination of spirochetes and that RpoS-mediated changes in the borrelial transcriptome collectively comprise a 'Go' signal for tick-to-mammal transmission. Additionally, we and others have identified a global regulatory network controlled by the Hk1/Rrp1 two-component system, which we conceptualize as imparting a tick- adaptive 'Stay' signal via the small nucleotide messenger cyclic di-GMP, a molecule known to promote sessile developmental states in bacteria. Our finding that the Hk1/Rrp1 regulon includes tick-phase genes repressed by RpoS within the mammal, coupled with our live-imaging studies, leads to our secondary hypothesis, to be assessed in Aim 3, that the interplay between RpoS and the Hk1/Rrp1 pathway distinguishes spirochete behavior during transmission from acquisition. The underlying premise of these studies is that improved understanding of the molecular mechanisms whereby Bb is transmitted by the arthropod vector will lead to new prevention strategies.
Borrelia burgdorferi (Bb), the causative agent of Lyme disease, is maintained in nature by an enzootic life cycle that involves an Ixodes tick vector and a mammalian host, typically wild rodents. We have shown that genes transcribed by the alternative sigma factor RpoS are critical for transmission of Bb by ticks as well as infection of mice and, therefore, humans. The experiments in our proposal will define more precisely how and where RpoS-dependent genes function during the transmission process and how they are regulated.
|Kenedy, Melisha R; Scott 2nd, Edgar J; Shrestha, Binu et al. (2016) Consensus computational network analysis for identifying candidate outer membrane proteins from Borrelia spirochetes. BMC Microbiol 16:141|
|Miller, Daniel P; Oliver Jr, Lee D; Tegels, Brittney K et al. (2016) The Treponema denticola FhbB Protein Is a Dominant Early Antigen That Elicits FhbB Variant-Specific Antibodies That Block Factor H Binding and Cleavage by Dentilisin. Infect Immun 84:2051-8|
|Caimano, Melissa J; Drecktrah, Dan; Kung, Faith et al. (2016) Interaction of the Lyme disease spirochete with its tick vector. Cell Microbiol 18:919-27|
|Bauer, William J; Luthra, Amit; Zhu, Guangyu et al. (2015) Structural characterization and modeling of the Borrelia burgdorferi hybrid histidine kinase Hk1 periplasmic sensor: A system for sensing small molecules associated with tick feeding. J Struct Biol 192:48-58|
|Iyer, Radha; Caimano, Melissa J; Luthra, Amit et al. (2015) Stage-specific global alterations in the transcriptomes of Lyme disease spirochetes during tick feeding and following mammalian host adaptation. Mol Microbiol 95:509-38|
|Caimano, Melissa J; Dunham-Ems, Star; Allard, Anna M et al. (2015) Cyclic di-GMP modulates gene expression in Lyme disease spirochetes at the tick-mammal interface to promote spirochete survival during the blood meal and tick-to-mammal transmission. Infect Immun 83:3043-60|
|Grassmann, AndrÃ© Alex; McBride, Alan John Alexander; Nally, Jarlath E et al. (2015) Generation of Mammalian Host-adapted Leptospira interrogans by Cultivation in Peritoneal Dialysis Membrane Chamber Implantation in Rats. Bio Protoc 5:|
|Bockenstedt, Linda K; Radolf, Justin D (2014) Xenodiagnosis for posttreatment Lyme disease syndrome: resolving the conundrum or adding to it? Clin Infect Dis 58:946-8|
|Caimano, Melissa J; Sivasankaran, Sathesh K; Allard, Anna et al. (2014) A model system for studying the transcriptomic and physiological changes associated with mammalian host-adaptation by Leptospira interrogans serovar Copenhageni. PLoS Pathog 10:e1004004|
|Kenedy, Melisha R; Luthra, Amit; Anand, Arvind et al. (2014) Structural modeling and physicochemical characterization provide evidence that P66 forms a *-barrel in the Borrelia burgdorferi outer membrane. J Bacteriol 196:859-72|
Showing the most recent 10 out of 25 publications