Kristine M. Wylie, Ph.D. - US grants

ABSTRACT An estimated 13 million preterm births (PTBs) occur annually worldwide, and PTB is the single most significant contributing factor to neonatal morbidity and mortality. Mechanisms underlying preterm birth are unknown hampering development of effective prediction and prevention strategies. PTB is linked to local and distant infections and recent data suggests that vaginal bacterial microbiome early in pregnancy is associated with subsequent preterm birth. Viral infections behave differently during pregnancy, with common viral infections such as varicella and influenza causing much more severe disease during pregnancy and human papillomavirus, the causative virus in cervical cancer, associated with a 2-fold increased risk for PTB. However, comprehensive assessment of vaginal viral communities during pregnancy has not been performed. Lastly, although maternal inflammation is one of the leading triggers for PTB the precise constellation of inflammatory signals is not known. We propose that examining microbial communities or host response alone is incomplete, but that their combination will lead to refined definitions of appropriate and inappropriate microbe-maternal biology and shed new light on the old problem of PTB. Our central hypothesis is that preterm birth can be estimated by a combination of three criteria: vaginal bacterial communities, vaginal eukaryotic viral communities, and maternal inflammatory response. We have an interdisciplinary team assembled including perinatologists, epidemiologists, virologists, and genomics informatics experts, leveraging the unique capabilities of the McDonnell Genome Institute at Washington University in St. Louis, a well-established pregnancy bio-specimen and clinical data infrastructure, and a high-PTB burdened racially diverse patient population to test this novel hypothesis. The aims of this project are: 1) Determine the ability of bacterial and viral communities in the vagina and their dynamics over time to predict preterm birth 2) Determine the ability of the host inflammatory response in the vagina in the context of microbial communities to predict preterm birth. The proposed research is innovative, for the first time, comprehensively characterizing the eukaryotic vaginal virome simultaneously with vaginal bacterial communities and longitudinally capturing the maternal response to these communities. We employ an efficient study design, a well-established research infrastructure, and renowned genome sequencing expertise to test a physiologically plausible but uninvestigated paradigm that bacterial and viral communities in concert with the maternal host response can predict PTB.

Project Summary There is currently no disease modifying treatment available for Alzheimer?s disease (AD) and our poor understanding of the factors that contribute to its development stands as a significant barrier to identifying effective preventative measures or treatments for this disease. This proposal seeks to advance our understanding of AD etiology by testing the hypothesis that infection with viruses that express inhibitors of protein phosphatase 2A (PP2A) can contribute to its development. Specifically, we will test a model wherein infection by PP2A inhibiting viruses could contribute to AD by triggering tau pathology that then propagates from infected cells to adjacent cells and brain regions in a pattern consistent with AD progression. This hypothesis is supported by three well-established lines of evidence from studies conducted in humans and animal models including: evidence demonstrating a role for PP2A in AD and tau pathology, the observation that tau pathology propagates to adjacent cells, and the fact that multiple human viruses express PP2A inhibiting proteins as a common strategy for coopting the physiology of their host cells. In this proposal, we will screen human brain tissue using a novel, state of the art method (ViroCap) and laser capture microdissection to identify and to test for associations between sites of viral infection and tau histopathology. This work will advance our understanding of the factors that contribute to the development of AD, with potential clinical applications including the development of tests to identify individuals at increased risk for AD as a result of infection with PP2A inhibiting viruses, the development of vaccines or antiviral therapies to prevent increased AD risk resulting from infection with these viruses, or the development of pharmacological interventions that reduce AD risk in infected individuals by increasing PP2A activity. In this proposal, we will also create an animal model of the seeding of tau pathology by viral PP2A inhibitors for use in understanding the mechanisms underlying their effects and identifying potential therapeutic approaches to combat them.

The U.S. has fallen behind several countries in STEM competiveness, where K-12 students are less proficient in science and math and earn fewer bachelor degrees in STEM than countries such as China and Korea. The maintenance of the U.S. economy relies heavily on innovation and technology. As well as our health systems rely on the discoveries of scientists and physicians. Thus STEM and health are critical to our nations welfare. Improving the education and STEM experiences for all students, but namely underrepresented minorities (URM) is also critical to our economy and healthcare as our country is projected to become a majority minority nation. We are proposing a program that will prepare low-income, URM high school students for careers in STEM and health. The Washington University Science Partnership Program will be housed at the McDonnell Genome Institute at Washington University in St. Louis and will partner with the Jennings School District, a largely African-American school district in St. Louis County. In this project we have proposed two aims to address the critical need for improved STEM education for URM. In Aim 1, we proposed to develop skills in these students in genomics and bioinformatics through authentic STEM experiences, both in class and through research internships. Science and health are approaching the era of personalized medicine due in part to the decoding of the human genome and the discoveries made as a result. However skilled persons in genomics and bioinformatics are in short supply compared to the demand and are especially lacking in URM groups. So this aim will seek to fulfill a direct need in science and prepare students for future, relatively lucrative jobs. In Aim 2 we will conduct workshops that focus on college and job readiness in STEM that will walk students through the process of applying to college as well as introduce them to the wide variety of career options in STEM. In total our proposed program is poised to increase the number of URM, namely African- Americans who enter college in STEM fields and who are retained in STEM and obtain careers in these fields. Our program directly addresses national priorities in STEM education: providing authentic research experiences to K-12 students, engaging underrepresented students in STEM and enhancing the STEM workforce.

ABSTRACT Preterm birth (PTB), delivery occurring before 37 weeks of gestation, affects approximately 10% of births in the United States each year and is the primary cause of neonatal morbidities and mortality. Because our understanding of the causes of PTB is incomplete, we are limited in our ability to predict or prevent PTB. In our previous studies, we used high-throughput sequencing to reveal characteristics of viral and bacterial communities in the vagina are associated with PTB. These data suggest that characteristics of the vaginal microbiome may serve as biomarkers for PTB, although more studies are required. While sequencing assays have been useful in beginning to identify microbial signatures associated with PTB, the incorporation of these assays into large-scale follow up studies is limited by several factors: (i) sequencing assays are in many cases qualitative, requiring subsequent assays to quantitate the microbes, thereby depleting sample material; (ii) sequencing and qPCR assays are time-consuming, requiring weeks to obtain complete data sets from each sample; and (iii) sequencing assays are relatively expensive despite many innovations that have reduced costs. Based on our increased understanding of vaginal microbial community features associated with PTB, we presently aim to develop a single, multiplexed, targeted assay that will allow us to capture the essential features we find in sequencing assays, but it will do so more quantitatively, rapidly, and cost-effectively than sequencing. The development of this assay will allow us (and others who study the microbiome of the human female reproductive tract) to maximize resource utilization in future studies by replacing multiple sequencing and PCR tests with a single, quantitative, rapid, easy to use, less expensive assay. Furthermore, this assay may serve as a foundation for future clinical studies aimed at defining risk for PTB in pregnant women

PROJECT SUMMARY Preterm birth and intrauterine growth restriction are significant public health problems in the United States, affecting 10% and 3-9% of births, respectively. These adverse pregnancy outcomes result in increased risk of mortality and lifelong morbidities, in addition to substantial monetary costs of >$26 billion annually. Intrauterine/placental inflammation contributes to more than half of preterm birth. Much of the research related to placental inflammation has focused on acute inflammatory processes, which are typically driven by bacterial infections in the amniotic cavity, and little is known about the triggers or mechanisms that initiate and sustain chronic placental inflammation (CPI). It is well known that some CPI is associated with viral infection, such as cytomegalovirus, but in most cases, the etiology of CPI remains unknown. We hypothesize that most CPI is a chronic inflammatory reaction to viral infection in the placenta. We will comprehensively analyze placental tissue from pregnant women to determine whether viruses and antiviral inflammatory profiles are associated with CPI. Furthermore, we will test maternal blood collected during pregnancy to determine whether we can identify a viral and/or host signal that will predict development of CPI. Lastly, we will test neonatal umbilical cord blood for the presence of viral infection or host signal consistent with that seen in the placenta or maternal blood. This study has the potential to lead to the development of a predictive or diagnostic test for CPI, and it will help us to understand how CPI is triggered and progresses so that we may ultimately be able to design a preventative or therapeutic treatment. Ultimately, our ability to predict and/or prevent CPI will improve pregnancy outcomes and child health.