Urinary Antimicrobial Peptides and the Urinary Microbiota in a UTI-Susceptible Population of Female Pelvic Floor Surgery Patients

Urinary tract infections (UTI)s are a national priority. Women who undergo surgery for pelvic floor disorders such as pelvic organ prolapse (POP) or urinary incontinence (UI) are at increased risk for UTI, as 10-30% will contract a post-instrumentation UTI (postI-UTI) within six weeks after surgery. Currently, the factors that contribute to the high rate of postI-UTI are unknown, and there is currently no clinical assessment to identify at-risk patients. However, antimicrobial peptides (AMPs) and characteristics of the urinary microbiota have the potential to serve as biomarkers, identifying patients at UTI risk and facilitating clinical prevention studies. While it was previously thought that the urinary tract was a sterile environment, new evidence shows that bacteria inhabit the urinary tract in many people. Given that microbial communities (microbiota) in other areas of the body exist in a balance with the host defense system, a similar equilibrium likely exists between the microbiota and host defenses in the urinary tract. AMPs, one significant component of our innate defense system, can limit pathogenic infection by their abilities to interact with and disrupt microbial membranes, and to stimulate immune cell recruitment. While it is reported that certain AMPs are expressed in the urinary tract, the AMP profile of the urinary tract has not been characterized. Inappropriate expression of AMPs in other tissues, including altered levels and/or activity, has been associated with several different disease states. It is therefore possible that POP/UI surgery patients that express inappropriate AMP levels or decreased AMP potency have an imbalance between their defenses and their resident microbiota, which results in their susceptibility to postI-UTI. These results begin to reveal the urinary antimicrobial peptide and microbiota profiles of three cohorts of patients: (1) POP/UI patients with culture-negative urine samples at baseline who DO NOT develop a postI-UTI (2) POP/UI patients with culture-negative urine at baseline who DO develop a postI-UTI, and (3) POP/UI patients with positive clinical cultures at baseline. While levels of two AMPs, psoriasin and human β-defensin-1, do not significantly differ between the cohorts, preliminary evidence suggests that a characteristic microbiota may exist in patients who develop infection. Furthermore, AMPs and the microbiota may directly influence one another. Levels of psoriasin are lower in patients with detectable E. coli in their urine than in patients with other types of infections. In addition to this correlation, the presence of certain genera of bacteria positively correlates with levels of psoriasin. This could indicate that some members of the microbiota, by affecting levels of AMPs, influence the susceptibility of the urinary tract to invading pathogens. By furthering our understanding of urinary AMPs and the urinary microbiota, we take an important step toward being able to identify POP/UI patients who are at high risk for postI-UTI, and toward developing new therapeutic options for preventing infection

Cutaneous Burn Injury Modulates Urinary Antimicrobial Peptide Responses and the Urinary Microbiome

OBJECTIVES: Characterization of urinary bacterial microbiome and antimicrobial peptides after burn injury to identify potential mechanisms leading to urinary tract infections and associated morbidities in burn patients. DESIGN: Retrospective cohort study using human urine from control and burn subjects. SETTING: University research laboratory. PATIENTS: Burn patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Urine samples from catheterized burn patients were collected hourly for up to 40 hours. Control urine was collected from "healthy" volunteers. The urinary bacterial microbiome and antimicrobial peptide levels and activity were compared with patient outcomes. We observed a significant increase in urinary microbial diversity in burn patients versus controls, which positively correlated with a larger percent burn and with the development of urinary tract infection and sepsis postadmission, regardless of age or gender. Urinary psoriasin and β-defensin antimicrobial peptide levels were significantly reduced in burn patients at 1 and 40 hours postadmission. We observed a shift in antimicrobial peptide hydrophobicity and activity between control and burn patients when urinary fractions were tested against Escherichia coli and Enterococcus faecalis urinary tract infection isolates. Furthermore, the antimicrobial peptide activity in burn patients was more effective against E. coli than E. faecalis. Urinary tract infection-positive burn patients with altered urinary antimicrobial peptide activity developed either an E. faecalis or Pseudomonas aeruginosa urinary tract infection, suggesting a role for urinary antimicrobial peptides in susceptibility to select uropathogens. CONCLUSIONS: Our data reveal potential links for urinary tract infection development and several morbidities in burn patients through alterations in the urinary microbiome and antimicrobial peptides. Overall, this study supports the concept that early assessment of urinary antimicrobial peptide responses and the bacterial microbiome may be used to predict susceptibility to urinary tract infections and sepsis in burn patients

Targeting deficiencies in the TLR5 mediated vaginal response to treat female recurrent urinary tract infection

Abstract The identification of the host defence peptides as target effectors in the innate defence of the uro-genital tract creates new translational possibilities for immunomodulatory therapies, specifically vaginal therapies to treat women suffering from rUTI, particularly those carrying the TLR5_C1174T SNP. Urinary tract infections (UTIs) are a microbial disease reported worldwide. Women are particularly susceptible with many suffering debilitating recurrent (r) infections. Treatment is by antibiotics, but such therapy is linked to antibiotic resistance and re-infection. This study explored the innate protective mechanisms of the urogenital tract with the aim of boosting such defences therapeutically. Modelling UTIs in vitro, human vaginal and bladder epithelial cells were challenged with uropathogenic Escherichia coli (CFT073) and microbial PAMPs including flagellin, LPS and peptidoglycan. Flagellin functioning via the TLR5/NFκB pathway was identified as the key UPEC virulence factor causing a significant increase (P < 0.05) in the production of the host-defence peptide (HDP), BD2. BD2-depleted urine samples from bladder infected mice supported increased UPEC growth, strengthening the significance of the HDPs in protecting the urogenital tissues from infection. Clinically, vaginal-douche BD2 concentrations were reduced (p < 0.05) in women suffering rUTIs, compared to age-matched healthy controls with concentrations further decreased (p < 0.05) in a TLR5392Stop SNP rUTI subgroup. Topical vaginal estrogen treatment increased (p < 0.001) BD2 concentrations in all women, including those carrying the SNP. These data identify therapeutic and antibiotic sparing roles for vaginal immunomodulatory agents that specifically target HDP induction, facilitate bacterial killing and disrupt the UPEC infection cycle

The urinary microbiome and its contribution to lower urinary tract symptoms; ICI-RS 2015

Aims: The microbiome is the term used for the symbiotic microbial colonisation of healthy organs. Studies have found bacterial identifiers within voided urine which is apparently sterile on conventional laboratory culture, and accordingly there may be health and disease implications. Methods: The International Consultation on Incontinence Research Society (ICI-RS) established a literature review and expert consensus discussion focussed on the increasing awareness of the urinary microbiome, and potential research priorities. Results: The consensus considered the discrepancy between findings of conventional clinical microbiology methods, which generally rely on culture parameters predisposed towards certain ‘‘expected’’ organisms. Discrepancy between selective culture and RNA sequencing to study speciesspecific 16S ribosomal RNA is increasingly clear, and highlights the possibility that protective or harmful bacteria may be overlooked where microbiological methods are selective. There are now strong signals of the existence of a ‘‘core’’ urinary microbiome for the human urinary tract, particularly emerging with ageing. The consensus reviewed the potential relationship between a patient’s microbiome and lower urinary tract dysfunction, whether low-count bacteriuria may be clinically significant and mechanisms which could associate micro-organisms with lower urinary tract symptoms. Conclusions: Key research priorities identified include the need to establish the scope of microbiome across the range of normality and clinical presentations, and gain consensus on testing protocols. Proteomics to study enzymatic and other functions may be necessary, since different bacteria may have overlapping phenotype. Longitudinal studies into risk factors for exposure, cumulative risk, and emergence of disease need to undertaken. Neurourol. Urodynam. 36:850–853, 2017

Envisioning and Enacting a Coherent Organization-Wide View of Data

Amidst the contemporary focus on open data and national coordination, there is a need for organization-centric discussions about scientific data and data management. Data collections at the National Center for Atmospheric Research (NCAR) and its parent organization the University Corporation for Atmospheric Research (UCAR) began in the mid-1960s, and have grown continuously, encompassing long-term reference data sets, field project data sets, and climate simulation results, among other data types. In addition, multiple NCAR and UCAR groups provide data tools and services. These wide-ranging efforts have largely been addressed as independent projects. While this has resulted in a large set of valuable services, it has also created challenges for coordinating efforts, sharing expertise, and presenting these services in a coherent fashion to potential users. Further, data services are not uniformly distributed around the organization, resulting in a spectrum of practices that varies among organizational units. As notions of ‘data access’ and ‘data integration’ develop, the work with data at NCAR and UCAR provides a case example of community-building activities and intra-organizational coordination over time. In this poster, we discuss four collective endeavors over the past 20 years to increase coordination and develop more coherence across UCAR and NCAR data services. Two of the recent efforts are ongoing. These undertakings are embedded within an evolving IT infrastructure so each is constrained by very different technologies. Though they differ in scope and outcomes, they share the overarching goal of increasing intra-organization coordination. We present lessons learned, including how prior efforts informed the creation of the current Data Stewardship Engineering Team activities within NCAR. This study will inform current open access and data sharing efforts by highlighting the importance of internal alignment and coordination within individual organizations. Evolution of an intra-organization view of data is a critical enabler of increased inter-organization data integration and discovery, and requires a continuing process of envisioning and re-envisioning.Data Curation Education in Research Centers (DCERC), IMLS Award# RE-02-10-0004-10Ope

Envisioning and Enacting a Coherent Organization-Wide View of Data

Amidst the contemporary focus on open data and national coordination, there is a need for organization-centric discussions about scientific data and data management. Data collections at the National Center for Atmospheric Research (NCAR) and its parent organization the University Corporation for Atmospheric Research (UCAR) began in the mid-1960s, and have grown continuously, encompassing long-term reference data sets, field project data sets, and climate simulation results, among other data types. In addition, multiple NCAR and UCAR groups provide data tools and services. These wide-ranging efforts have largely been addressed as independent projects. While this has resulted in a large set of valuable services, it has also created challenges for coordinating efforts, sharing expertise, and presenting these services in a coherent fashion to potential users. Further, data services are not uniformly distributed around the organization, resulting in a spectrum of practices that varies among organizational units. As notions of ‘data access’ and ‘data integration’ develop, the work with data at NCAR and UCAR provides a case example of community-building activities and intra-organizational coordination over time. In this poster, we discuss four collective endeavors over the past 20 years to increase coordination and develop more coherence across UCAR and NCAR data services. Two of the recent efforts are ongoing. These undertakings are embedded within an evolving IT infrastructure so each is constrained by very different technologies. Though they differ in scope and outcomes, they share the overarching goal of increasing intra-organization coordination. We present lessons learned, including how prior efforts informed the creation of the current Data Stewardship Engineering Team activities within NCAR. This study will inform current open access and data sharing efforts by highlighting the importance of internal alignment and coordination within individual organizations. Evolution of an intra-organization view of data is a critical enabler of increased inter-organization data integration and discovery, and requires a continuing process of envisioning and re-envisioning.Data Curation Education in Research Centers (DCERC), IMLS Award# RE-02-10-0004-10Ope

NCAR_DSET_poster-ESIP-2018.pdf

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