More-WH Scholars

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BIRCWH Scholars

The Women's Health Research Group, UMB appointed our first MORE-WH Scholar, Jenny Jones, Ph.D., Research Associate, Department of Physiology, School of Medicine, on January 1, 2003. Our second, third and fourth Scholars, William Romani, Ph.D., P.T., A.T.C., S.C.S., Assistant Professor, Department of Physical Therapy & Rehabilitation Sciences, School of Medicine, Jessica A. Mong, Ph.D., Assistant Professor, Department of Pharmacology & Experimental Therapeutics, School of Medicine, and Vasana Cheanvechai, M.D., Assistant Professor, Division of Vascular Surgery, Department of Surgery, School of Medicine, were appointed July 1, 2003. On January 1, 2005, we appointed our fifth and sixth Scholars, Leonardo Tonelli, Ph.D., Assistant Professor, Department of Psychiatry, School of Medicine, and Kathleen Tracy, Ph.D., Assistant Professor, Department of Epidemiology & Preventive Medicine, School of Medicine. Two more Scholars, Laundette P. Jones, Ph.D., Assistant Professor, Department of Pharmacology & Experimental Therapeutics, School of Medicine and Gregory Hicks, M.P.T, Ph.D., Assistant Professor, Department of Physical Therapy and Rehabilitation Sciences, School of Medicine were appointed July 1, 2005. Our final two Scholars for this award, Tami Kingsbury, Ph.D., Department of Physiology, School of Medicine and Sharon Gordon, D.D.S., M.P.H., Ph.D., Associate Professor, Department of Biomedical Sciences, Dental School were appointed on January 1, 2006. Individuals interested in future appointment opportunities should contact:

Patricia Langenberg, Ph.D., Principal Investigator
Department of Epidemiology & Preventive Medicine
University of Maryland, Baltimore
Room 102A, 660 W. Redwood Street
Baltimore, Maryland 21201
(410) 706-3251
plangenb@umaryland.edu

Istvan Merchenthaler, M.D., Ph.D., D. Sc., Program Director
University of Maryland, Baltimore
Department of Epidemiology & Preventive Medicine
Room 9-00F, 685 W. Baltimore Street
Baltimore, Maryland, 21201
(410) 706-1350
imerchen@epi.umaryland.edu

Jenny M. Jones, Ph.D.
Appointment date: 1/1/2003 - 8/31/2005

Statement of Research Plan

I. Mentor Team

Robert Koos, Ph.S., Professor, Department of Physiology, School of Medicine (Lead Mentor)
Angela Brodie, Ph.D., Professor, Department of Pharmacology & Experimental Therapeutics, School of Medicine
Jodi Flaws, Ph.D., Associate Professor, Department of Epidemiology & Preventive Medicine, School of Medicine
Katherine H. Tkaczuk, M.D., Associate Professor, Departments of Medicine and Oncology, School of Medicine

II. Introduction

Mechanisms involved in breast cancer cell growth, invasion, and metastasis are in many ways similar to those involved in the normal wound healing process, e.g. cell proliferation, migration, epithelial-stromal cell interaction, cell-matrix interaction, and upregulation of growth factors, adhesion molecules, and matrix-degrading proteases. Because of her background in the area of cutaneous wound healing, Dr. Jones has become very interested in these processes in breast cancer cells. Although much is known about some of these processes in breast cancer, far less is known about the regulation of specific gene expression by components of the extracellular matrix (ECM), or about the signaling pathways involved in this regulation. Dr. Jones plans to determine how different ECM components regulate expression of relevant genes in normal mammary epithelial cells and in breast cancer cells. These genes include plasminogen activators and their inhibitors, ER±, ER², relaxin and its receptors, progesterone receptor, and integrins. This research will increase our understanding of how the ECM controls the expressions of these genes and will have a significant impact in the area of breast cancer research.

III. Proposed Project

Regulation of Plasminogen Activation System Components in Breast Cancer

Hypothesis
Gene expression of components of the plasminogen activation system, specifically PAI-1 and tPA, are controlled by the extracellular matrix in skin epithelial cells (Jones et al., Exp. Cell Res. 280:244-254, 2002). PAI-1 is highly expressed in breast cancer cells, and is a poor prognostic indicator, while tPA is a favorable prognostic indicator. Therefore, Dr. Jones plans to investigate the hypothesis that the ability of ECM components to regulate PAI-1 and tPA gene expression is different in normal vs. malignant mammary epithelial cells.

Specific Aims

Determine expression of PAs and PAI-1 in normal mammary epithelial cells in response to treatment with ovarian hormones in the context of different ECM proteins.
Identify components of the signaling pathway(s) (i.e. integrins, kinases, response elements) that are involved in regulation of PA and PAI-1 expression by these ECM proteins in these cells.
Determine whether differences exist in ECM regulation of PAI-1 gene expression in normal mammary cells vs. breast cancer cells. If differences are found, investigate the bases for the differences.
Manipulate relevant signaling pathway components by transfection or mutation studies to prove cause and effect.

William Romani, Ph.D., P.T., A.T.C., S.C.S.
Appointment date: 7/1/2003-6/30/2005

Statement of Research Plan

I. Mentor Team

Jodi Flaws, Ph.D., Associate Professor, Department of Epidemiology & Preventive Medicine, School of Medicine (Lead Mentor)
Robert Koos, Ph.D., Professor, Department of Physiology, School of Medicine
Angela Brodie, Ph.D., Professor, Department of Pharmacology & Experimental Therapeutics, School of Medicine

II. Proposed Project

Dr. Romani's long-term research objective is to identify the mechanisms of sex hormones influence on collagen remodeling in order to provide a scientific basis for women's increased risk for ACL injury. Recent Program Announcements in the area of Women's Health and Exercise that are sponsored by NIAMS and the Office of Research on Women's Health suggest that these research questions are fundable. The hypothesis of this proposal is that the ligand activated transcription of fibroblasts, procollagen, and metalloproteinases that modulate the tensile strength of the ACL are regulated by the expression of estrogen receptors. Dr. Romani expects that his findings will allow him to further investigate the role of ligand receptors, including those for progesterone.

Hypothesis
The central hypothesis is that the ligand activated transcription of fibroblasts, procollagen, and metalloproteinases that modulate the tensile strength of the ACL are regulated by the expression of estrogen receptors.

Specific Aims

Establish the acute effects of estradiol and progesterone concentration on ER± and ER² expression in rat ACL tissue. To accomplish this aim, six groups of female Sprague-Dawley rats will be ovariectomized and injected with estradiol, progesterone, or a combination of these on one or two successive days. A seventh control group will receive an injection without either sex hormone. Dr. Romani will quantify ER± and ER² mRNA expression in the seven groups with semi-quantitative PCR. He expects that the rats injected with estradiol will demonstrate increased ER± and decreased ER² mRNA expression in ACL tissue, that progesterone injection will result in decreased ER± and increased ER² mRNA expression, and that injections of estradiol prior to progesterone and progesterone prior to estradiol will result in decreased expression of ER² and increased expression of ER±, respectively.
Identify the changes in ER± and ER² mRNA expression in rat ACL tissue throughout a normal estrous cycle. Dr. Romani hypothesizes the ER± mRNA expression will increase and ER² mRNA expression will decrease during the late proestrus (day 3) phase of the estrous cycle and that ER± mRNA expression will decrease and ER² mRNA expression will increase and during early estrous (day 4). To demonstrate this, Dr. Romani will use semi-quantitative PCR to determine relative levels of ER± and ER² mRNA expression in the ACL of rats at three phases of the estrous cycle.

Jessica A. Mong, Ph.D.
Appointment date: 7/1/2003 - 2/28/2006

Statement of Research Plan

I. Mentor Team

Angela Brodie, Ph.D., Professor, Department of Pharmacology & Experimental Therapeutics, School of Medicine (Lead Mentor)
Patricia Langenberg, Ph.D., Professor, Department of Epidemiology & Preventive Medicine, School of Medicine
Margaret M. McCarthy, Ph.D., Professor, Department of Physiology, School of Medicine

II. Proposed Project

In recent years, the well-established tenets of physiology have begun to be re-examined in the context of the female body and striking differences are emerging. The female endocrine system lies at the heart of these differences with estrogens being key ovarian steroids affecting these differences. Estrogens have wide-ranging effects in the central nervous system (CNS) of most adult mammals including humans and have been reported to enhance cognitive functions as well as serve a neuroprotective role. However, conflicting reports have made these studies controversial at best. In spite of the knowledge that estrogens regulate the expression of a number of different genes in the CNS, we lack a complete understanding of estrogen-targeted genes in the CNS and how changes in their transcript levels may contribute to the myriad of neurobiological functions affected by estrogens. Dr. Mong's general research interests evolve around the molecular and cellular mechanisms underlying estrogenic actions in the CNS. More specifically, she is interested in the cellular and molecular mechanisms of estradiol mediated neuronal-glial interactions and her current findings suggest a molecular pathway in which estradiol my be affecting sleep, a frequent problem for post-menopausal women.

There is a growing appreciation for the importance of glia, a heterogeneous group of non-neuronal cells in the brain, to overall synaptic functioning. In vivo, the nature of neuronal-glial interactions is an intimate one and as such presents a challenge when attempting to study the molecular mechanisms underlying estrogenic effects. However, the development and continued improvement of DNA microarray technology has facilitated the simultaneous assessment of the expressions of thousands of genes, both neuronal and glial. In her post-doctoral position at the Rockefeller University, Dr. Mong successfully used microarray technology to elucidate the short- and long-term actions of estrogens on the transcriptions of several estrogen-concentrating hypothalamic nuclei. The results have demonstrated a number of unanticipated gene regulations that include many glial-specific genes. One of the more interesting ones is prostaglandin D synthase (L-PGFS), a non-neuronal enzyme that catalyzes the conversion of Prostaglandin H2 to Prostaglandin D2(PGD2) a neuromodulator that is involved in a variety of functions including sedation and sleep. Subsequent confirmation using the high spatial resolution technique on in situ hybridization revealed an intriguing differential regulation of the L-PGDS mRNA transcripts. In the arcuate and ventromedial nucleus of the hypothalamus, estradiol increases expression but it dramatically suppresses expression in the ventolateral preoptic area (VLPO), a putative sleep center in the brain. In women as well as female rodents, fluctuations in circulating levels of estrogens have been shown to effect sleep homeostasis. In light of this discovery in the VLPO, it is tempting to speculate that a subsequent decrease in the PGD2 may contribute to the general arousal mediated by estrogens and suggests a molecular mechanism for how estrogens may be affecting sleep in mammals.

Hypothesis 1: Estradiol regulation of L-PGDS expression involves neuronal-glial interactions.

Specific Aims: Hormone mediated neuronal-glial interactions in the expression of L-PGDS

Determine cell types synthesizing L-PGDS in the VLPO and VMN.
Further characterize the estradiol mediated neuronal-glial interaction controlling L-PGDS promoter activity.
Identify key elements in the L-PGDS promoter that are key in mediating estradiol action.

Hypothesis 2: Hormonal regulation of L-PGDS expression affects general arousal and/or sleep-wake cycles.

Specific Aims:

Further characterize hormonal responsiveness of L-PGDS expression:
- specificity (Testosterone, Estradiol+Progesterone)
- dose response and pharmacological inhibition
- hormonal regulation in other brain regions (e.g. amygdala)
Role of estradiol mediated expression of L-PGDS in the VLPO in sleep-wake cycles.
Role of hormone mediated neuronal-glial interactions in increasing general arousal states.

Vasana Cheanvechai, M.D.
Appointment date: 7/1/2003 - 10/30/2005

Statement of Research Plan

I. Mentor Team

Jodi Flaws, Ph.D., Associate Professor, Department of Epidemiology & Preventive Medicine, School of Medicine (Lead Mentor)
Julie Ann Freischlag, M.D., Professor & Chair, Department of Surgery, Johns Hopkins University
Patricia Langenberg, Ph.D., Professor, Department of Epidemiology & Preventive Medicine, School of Medicine

II. Proposed Project

Adverse Conditions and Diseases in Women: Noncardiac Vascular Disease

Dr. Cheanvechai's work in vascular disease research began as a medical student analyzing ventricular relaxation in an animal model. During her surgical residency, she spent a year at Northwestern University where she examined the expression of matrix metalloproteinases in carotid artery plaques. As a Fellow in the Vascular Surgery at the University of Maryland School of Medicine, Dr. Cheanvechai continued the evaluation of the role of MMP expression in vascular remodeling and collateral vessel formation in a rabbit hindlimb ischemia model. During this Fellowship, she also completed several clinical research projects regarding vena cava filters and lower limb revascularization procedures.

In addition to her appointment as Assistant Professor of Surgery in the Division of Vascular Surgery, Dr. Cheanvechai also serves as Medical Director of the Non-Invasive Vascular Laboratory at the University of Maryland Hospital. Recently she has been conducting noninvasive screening programs to detect noncardiac vascular disease. Through these types of screening programs, both locally and nationally, Dr. Cheanvechai plans to study the incidence and prevalence of noncardiac vascular disease, especially as it pertains to women. She hopes to increase public awareness of and public education about noncardiac vascular disease and to reduce deaths and disability due to those disorders. She believes the ability to perform noninvasive screening programs will help to provide a unique opportunity to study noncardiac vascular disease in women in this country, and help to develop national public education programs in women's vascular health that may result in a significant improvement in detection and treatment of these problems and ultimately, a reduction in adverse cardiovascular events in women.

Vascular Disease in Women's Health

The critical clinical importance of cardiovascular disease in this country grows exponentially with each decade of our aging population. Most people are well aware of the importance of prevention, detection, and treatment of heart disease; but few people are aware of the seriousness of noncardiac vascular disease, other than those who have suffered its complications. Noncardiac vascular diseases such as carotid artery disease, peripheral arterial disease (PAD), and abdominal aortic aneurysms (AAA) combined, cause almost as much death and disability in this country as heart disease and more than any cancer. Yet these diseases with their associated risks of stroke, limb loss and death are undiagnosed and untreated in the majority of cases, even in groups at obvious high risk for atherosclerotic disease.

While it may seem implied that the importance of heart disease is ubiquitous, it has become evident in the last decade that even heart disease has been under-diagnosed and under-treated among women in this country. Considering what we clearly know about the lower levels of recognition of noncardiac vascular disease compared to heart disease, it is unimaginable that this same diagnostic and therapeutic chasm between men and women does not also exist for noncardiac vascular diseases. Further, while it has generally been accepted that the prevalence of vascular disease is greater in men than women, we have observed that this gap narrows progressively with increasing age. Like heart disease in women, noncardiac vascular disease in women must become a national health priority.

BACKGROUND: Noncardiac Vascular Disease

Carotid Artery Disease. Stroke is the third leading cause of death in the United States and a leading cause of permanent disability among older Americans. Almost 80% of strokes are ischemic, due to cerebrovascular disease that occurs most often in the carotid arteries. It is estimated that over $70 billion is spent each year in this country in the care of stroke victims. People with symptoms of cerebrovascular disease are aggressively evaluated but more than a third of stroke victims have no symptoms prior to their disabling stroke. When critical carotid artery disease is detected, most people can receive treatments that will significantly reduce the risk of stroke. Duplex ultrasound scanning can provide accurate diagnosis of carotid artery disease in a matter of minutes, without risk or discomfort, but programs for screening well recognized high-risk patient groups have not previously been developed. The tremendous burden of stroke, both personal and national, suggest that we should aspire to a healthcare system that can effectively focus its resources more on stroke prevention than on stroke survivors.

Peripheral Arterial Disease. Peripheral arterial disease effects 10-20% of people over the age of 70 in this country, a population group that could reach 20-30 million Americans in the next decade. PAD may produce difficulty walking (claudication) and can worsen and require invasive treatments (angioplasty or bypass) to prevent amputation. Overall though, the majority of people with mild to moderate PAD have few symptoms. However, it is now well recognized that people with PAD have a 2-3 fold increase in the incidence of heart attack and stroke compared to those without PAD. The majority of people with PAD will not require invasive treatments for their limbs, but it has become apparent that all people with PAD will require invasive treatments designed to reduce adverse cardiovascular events (e.g. smoking cessation, blood pressure control, lipid lowering regimens, anti-platelets medications, beta-blockers, etc.) It appears intuitive that, like heart disease, earlier detection of PAD will allow the effective use of these lower-risk interventions. PAD can be easily diagnosed in minutes by a Doppler-assisted measurement of the ankle-brachial index (ABI). While the ABI is part of the routine exam performed by vascular specialists, it has not been performed routinely by the majority of primary care providers in this country.

Abdominal Aortic Aneurysm. Abdominal aortic aneurysm is the 10th leading cause of death in men over age 55, and at least 15,000 deaths occur each year in this country due to ruptured AAA. Men generally outnumber women by a ratio of 4-6:1; but the overall risk of AAA increases for both sexes with age. As our population has aged, this wide difference in prevalence between men and women has already been observed to narow significantly, but the risk of aneurysm rupture remains the same. When larger aneurysms are diagnosed early, they can be treated safely, many today using less invasive techniques that avoid the need for major surgical procedures. However, when aortic aneurysms go on to rupture they are fatal in more than half the cases. AAA can be easily diagnosed in minutes without risk or discomfort using duplex ultrasound scanning. However, even among high risk patient groups (e.g. elderly hypertensive men who are active smokers, or those with family members with AAA) there have not been active screening programs for AAA in this country.

RESEARCH GOALS

Dr. Cheanvechai believes that a broad-based community outreach screening program could be an effective vehicle for promotion of public awareness about vascular disease. Vascular surgeons have pioneered and progressively refined the use of noninvasive vascular testing with Doppler and duplex ultrasound scanning for major vascular disorders, so a screening initiative would be an application of well established and accepted technology. In 2002, the American Vascular Association (AVA) performed its first annual National Screening Program for carotid disease, AAA and PAD, in older Americans at high risk for atherosclerosis. Over 850 people were tested in the first year of this program, and over 60% of those who presented for screening were women, and over 13% of people tested had abnormal exams.

More than 7% of people screened in 2002 were found to have PAD (ABI < 0.85) but less than half of these people were receiving medical treatments (e.g. anti-platelet agents) that might reduce their well recognized risks of heart attack and stroke. Almost 8% of people tested had carotid artery disease (ICA stenosis >50%) and 2% had previously undiagnosed critical carotid stenosis (80-99%). Like those with PAD, less than half the people with carotid stenosis were receiving antiplatelet medications. Additionally, among people with PAD and those with carotid disease, less than half were receiving lipid-lowering medication. Aortic aneurysms were found in only 2% of the population screened, but 20% of the AAA detected were over 6 cm diameter and had not been previously disgnosed; arguably life-saving events in these cases.

Overall this screening program appeared to confirm that vascular disease is underdiagnosed and under-treated in a broad segment of the population nationwide. These findings clearly support efforts to develop a national public education program regarding noncardiac vascular disease, as well as to learn more about its prevalence and distribution in different populations. These findings also suggest an opportunity for physician education regarding vascular disease, particularly among our primary care-givers.

Additional large scale screenings will provide a wealth of information that will progressively elucidate the true prevalence of vascular disease in women compared to men, as well as differences in current treatment and prevention patterns in women. Further, demographic data obtained will help to identify specific high-risk age, ethnic and geographic groups that will allow refinement of screening recommendations to increase the impact of more programs in the future. Such information is not currently available about vascular disease in women, and effective public and physician education (focusing on early preventive measures) will provide an opportunity to improve outcomes. An effective national education program about vascular disease in women, like the one currently being implemented for heart disease (the HeartTruth - NHLBI/AHA) should result in earlier diagnosis, broader and more effective preventative treatments, and ultimately in a measurable decrease in adverse cardiovascular events such as myocardial infarction, stroke, and aneurysm rupture. The success of such a program would improve the quality of life among older women in this country, and significantly reduce health care expenditures.

Dr. Cheanvechai plans to establish local screening programs for detection of vascular disease in women in the Baltimore City and Baltimore County region. These local screenings will provide information about true prevalence of vascular disease in women compared to men, as well as differences in current treatment and prevention patterns in women. In the subset of patients identified with vascular disease, further studies such as lipid profiles and CRP will be done in order to institute risk factor modification to improve outcomes.

As more screenings are conducted, Dr. Cheanvechai will attempt to accomplish some of the following goals:

Determine the prevalence of major vascular disease among women in this country:
Current available epidemiological data about noncardiac vascular disease in women is woefully inadequate to predict the true prevalence of these disorders, particularly in the complex subgroups of our population. Information about preventative treatments being afforded to women in such cases is almost non-existent.
Develop a national education program about vascular disease in women:
Public education-Older Americans are actively seeking healthcare information in greater numbers, using the Internet and other resources. Women lead the way in these activities and it is well recognized that women are more likely to facilitate healthcare decisions for their spouses and other family members (particularly older parents and relatives). It would be expected that women would respond favorably to such a program and would become proactive in discussions with healthcare providers regarding these issues.
Physician education-In a short number of years, an effective program would be expected to demonstrate a significant increase in preventative treatments being afforded to women with known vascular disease or those in then well-documented high-risk groups.
Refine geographic and demographic criteria for healthcare cost-effective screening for vascular disease in higher risk women's group:
Within a short number of years analysis of the results of this national screening program will allow us to specifically identify the highest risk women's subgroups. This will allow more specific recommendations regarding screening for these different groups. This will also facilitate the development of specific recommendations regarding vascular disease screening to move forward to the federal level.
Provide simpler, more effective early detection programs for medically under-served, high-risk populations:
Populations that are medically under-served, whether by virtue of geography, economy, or ethnicity, are more likely to present for care with advanced disease and frequently after adverse events. These people are more likely to require more invasive treatments and thus more costly and complex care. Historically these populations have been resistant to attempts at preventative care, efforts that are often viewed as culturally invasive. I believe the information from screening programs will provide a compelling message for community leaders about the risks of vascular disease in their own communities, and encourage them to support these programs locally. Further, women often dominate the family unit in these populations. Finally, these noninvasive screening techniques are universally acceptable because of their simplicity, and these programs can be performed easily in a community setting.
Develop parameters for assessment of the effectiveness of the screening/public education program:

Increased use of antiplatelet therapy in women with carotid artery disease.
Increased use of lipid-lowering therapy in women with PAD or carotid disease.
Reduced incidence in ruptured AAA among women.

Leonardo Tonelli, Ph.D.
Appointment date: 1/1/2005-12/31/2006

Statement of Research Plan

I. Mentor Team

Joel Greenspan, Ph.D., Professor, Department Biological Sciences, Dental School (Lead Mentor)
Istvan Merchenthaler, M.D., Ph.D., D.Sc., Professor, Departments of Epidemiology & Preventive Medicine and Anatomy & Neurobiology, School of Medicine
Margaret M. McCarthy, Ph.D., Professor, Department of Physiology, School of Medicine

II. Background

Epidemiological studies suggest that women may be more susceptible to the development of chronic inflammatory diseases. Women develop rheumatoid arthritis, systemic lupus erythematosus and multiple sclerosis 2.5, 6, and 2 times more frequently than do men, respectively. The pathogenesis of these diseases involves inflammatory and immune responses, which are believed to be associated with female-specific hormonal environment factors. The mechanisms by which sex hormones relate to these inflammatory diseases are poorly understood. However, the glucocorticoid hormonal system interacts both with sex hormones and with inflammatory reactions suggesting that it is relevant to the understanding of the relationship between hormonal environment and inflammatory/autoimmune response.

Dr. Tonelli used an animal model based on genetically determined differential responses to stress to study the relationship between female sex hormones, immune responses and inflammation. Inbred female Fischer and Lewis rat strains had either hyper- or hypo-responsive glucocorticoid and hypothalamic pituitary adrenal (HPA) axis responses. He challenged the immune system of these animals to study estrogen receptor regulation in the HPA axis. The main finding of this study was that the strain of rats susceptible to inflammation and which displayed a defective response to stress had low estrogen receptor responses. A second important finding was that transcriptional expression of estrogen receptor was changed even though estrogen concentrations were stable during the immune challenge. These experiments suggest that estrogen receptors are capable of being regulated by factors other than estrogen during an inflammatory challenge.

In a second study Dr. Tonelli used the same strains of rats to analyze the expression of inflammatory genes in brains of female rats. The purpose of this study was to evaluate the relationship between differential stress responsiveness and the brain's inflammatory response. Dr. Tonelli found that animals with a high stress response and a high level of resistance to development of inflammation had strong initial inflammatory gene expression. He concluded that these findings suggest that a strong expression of inflammatory genes may be necessary during the initial phase to prevent propagation of inflammation.

Dr. Tonelli used this model to extend his investigations to examine the role of early life experience on adult responses to stress. By cross-fostering the pups of these two strains of rats, he was able to evaluate the contribution of maternal behavior in the maturation of the stress and inflammatory responses in male and female rats of each strain. In both strains of rats, females were more resistant than males to the effects of cross-fostering as measured by adult behavioral responses to stress and to glucocorticoid responses to immune challenge. Females showed reduced adult glucocorticoid responses to stress related to the cross-foster procedure; males showed the opposite effect. In addition, these studies showed that the inflammatory response in the susceptible strain is not changed by maternal behavior. In summary, these studies suggest that resistance to inflammation in females is assoicated with a strong initial inflammatory and stress response and that this response has an important genetic component.

III. Goals

In a recent collaborative study with members of the Mood and Anxiety Program at the Department of Psychiatry, University of Maryland, Dr. Tonelli found an association between the peak rate of occurrence of suicide in women, age 15 to 45, and the spring peak concentration of atmospheric tree pollen. This association between rate of suicide and atmospheric tree pollen did not occur for men. Dr. Tonelli's has hypothesized that seasonal allergies triggered by tree pollen effect mood by release of inflammatory mediators thereby rendering vulnerable individuals susceptible to suicide. Dr. Tonelli's findings are consistent with a recent report stating that suicide occurrence was associated with season only in atopic individuals. Three other recent studies have described women-only associations between allergy and depression. This association between allergies and depression is poorly understood. Dr. Tonelli's goal is to determine mechanisms by which activation of the immune response (either by allergies or by infection) triggers depression and anxiety in vulnerable women. He expects to obtain basic information about the effect of the brain's immune response on hormonal systems, how hormonal systems affect the brain's immune response, and how these factors regulate brain functions, including emotion and cognition. Because of the relationship between chronic inflammation, immune response, and hormonal environment, this research focus is germane to the understanding of the group of diseases of unknown cause that disproportionately affect women. Post-partum depression, a disease associated with high costs in suffering and human productivity, serves as a salient example.

IV. Research Interests in Women's Health

To advance the understanding of risk factors in the development of mood and anxiety disorders in women, Dr. Tonelli plans to study the molecular mechanisms of hormonal regulation in the brain. He hopes to understand the neurological basis of the association between activation of immunity and the pathogenesis of mood and anxiety disorders in women. A rat animal model will be used to achieve these objectives. In particular, Dr. Tonelli will compare the immune response of male and female rats under physiological conditions and after changing their sex hormone environment. At the same time, he will evaluate the effects that these factors have on mood and anxiety.

Three inflammatory system challenge paradigms will be used: 1) challenge with bacterial lipopolysaccharides (LPS) to activate toll-like receptor 4 mediated brain innate immune response, 2) mimic anti-viral and anti-bacterial genomic DNA product activation of anti-viral and anti-infectious response (as mediated by toll-like receptors 7 and 9) using CpG oligonucleotides, 3) sensitization and re-challenge with tree pollen to activate the TH2 anti-parasitic response and the allergic response mediated by B cells and mast cells. In all cases, after the challenges, the animals will be tested using the forced swimming test (as a measure of depression) and the elevated plus maze (as a measure of anxiety). To determine the effect of female sex hormones on behavior, animals will be castrated, given sex hormones and retested behaviorally. The behavioral performance will be compared between sexes and between control and immune challenged animals. Female animals will be evaluated during different phases of the estrous cycle. At the least, Dr. Tonelli expects to determine dimorphic response to the behavioral test in the immune challenged group.

Each animal will be tested under only one condition. After behavioral testing, the animals will be sacrificed. The brains will be removed and dissected for mRNA extraction. The following brain structures will be processed for mTNA quantification: prefrontal cortex, hypothalamus, and brain stem. For each brain region corresponding to one of the experimental treatments (control, immune challenged, hormone replaced, etc.) real time RT-PCR gene analysis will be used to quantify expression of the following pro-inflammatory and anti-inflammatory genes: interleukin-1 beta, tumor-necrosis factor alpha, interleukin-6, interferon gamma, interleukin-4, interleukin 10, and interleukin 12. Focusing on gender differences, Dr. Tonelli expects to find differential regulation of at least one of the proposed candidate genes in females as compared to males.

Dr. Tonelli expects that the information obtained from these studies will help to determine if there is a sex-related inflammatory and behavioral response in the brain that is associated with the higher prevalence of mood and anxiety disorders in women. These experiments will establish the basis for future experimental and interventional studies targeting specific components of the inflammatory response in specific brain regions of female rats. This information should increase the understanding of the role of allergies and other immune system disorders in the increased incidence of mood and anxiety disorders in women. Finally, it is hoped that information obtained as a result of these studies will provide new therapeutic and preventive strategies to treat women (and men) suffering from mood and anxiety disorders.

J. Kathleen Tracy, Ph.D.
Appointment date: 1/1/2005 - 6/30/2007

Statement of Research Plan

I. Mentor Team

Renee Royak-Schaler, Ph.D., M.Ed., Associate Professor, Department of Epidemiology & Preventive Medicine, School of Medicine (Lead Mentor)
Jodi Flaws, Ph.D., Associate Professor, Department of Epidemiology & Preventive Medicine, School of Medicine
Patricia Langenberg, Ph.D., Professor, Department of Epidemiology and Preventive Medicine, School of Medicine

II. Summary of Academic and Research Background

Dr. Tracy's research interests have evolved over time as a function of her academic training, work experience, professional interactions, and personal philosophy and interests. During her undergraduate training and the early part of her graduate training, her interests were primarily focused on pediatric/developmental research topics (e.g. adjustment following pediatric bone marrow transplantation, treatment compliance among children with cystic fibrosis, correlates of attachment). However, as her graduate training continued and she began to work in applied settings, she developed a greater interest in working with adult populations. While completing her doctoral degree, she worked under the supervision of Dr. Robert McCarter and expanded her research skills in the areas of epidemiology, applied informatics and statistical analysis by working as an informatics consultant and statistical analyst on numerous biomedical research projects, many of which involved a gender-based focus. It was in her capacity as informatics and statistical analysis support personnel that her interest in gender-based medicine and women's health research began to emerge as focus areas of research interest. She was fortunate to have the opportunity to collaborate with several senior researchers at the University of Maryland whose line of research included a focus on gender as an important factor in disease incidence and outcome.

In late 2001, Dr. Tracy attended the University of Maryland's Women's Health Research Group-sponsored symposium on women's sexuality. While attending the conference, she took note of the fact that there was an absence of content related to geriatric women, lesbians, and other groups of women who are frequently underserved or under-represented in women's health research. This led her to review the current state of the science related to women's sexual functioning among some of these groups and the discovery that there is a paucity of research on what might be characterized as "marginalized" groups of women and sexual functioning. Consequently, she elected to complete a dissertation project that would be a first step toward closing the knowledge gap for at least one of these groups of women--lesbians--and began to actively focus her own research activities on women's health research.

Dr. Tracy received her Ph.D. in Psychology from the University of Maryland Baltimore County in August, 2003. During her graduate training, she specialized in health psychology and behavioral medicine and focused her doctoral research on correlates of sexual functioning among lesbians. Upon completion of her doctoral degree, she accepted a faculty position with the Department of Epidemiology and Preventive Medicine (DEPM) at the University of Maryland School of Medicine, and in July of 2004, she was promoted from the rank of Instructor to the rank of Assistant Professor, tenure track. She joined DEPM as a member of the Biostatistics and Bioinformatics division where she currently serves as an informatics consultant and collaborator on several research projects with a gender-based or women's health focus (e.g. interstitial cystitis, osteopororsis in men). In 2004, she also accepted joint affiliation with the division of Gender-based Epidemiology with the hope of cultivating collaborations that would support her women's health research agenda.

III. Professional Goals/Objectives

In collaboration with her faculty mentor, Dr. Jodi Flaws, Dr. Tracy has developed a set of professional goals that she believes are well suited to a successful career in women's health research and will provide a strong basis for her academic career at the University of Maryland School of Medicine. Dr. Tracy's primary professional goals are to

establish herself as an independent researcher in the area of women's health with a specific research agenda that focuses on cancer prevention and cancer survivorship issues among women; the specific emphasis of her research agenda will be under-served and under-represented women who are at unique risk of developing cancer or who may have unique issues related to survival following cancer diagnosis;
obtain and maintain on-going grant support to further her research agenda;
establish a national reputation in her chosen area of women's health research via scholarly publications, presentations, and participation in peer review activities;
grow a successful and diverse research team to support her women's health research activities and to provide an environment in which to mentor others (graduate students, fellows) in women's health research;
continue her teaching efforts in women's health and applied informatics.

The specific objectives designed to help Dr. Tracy achieve these goals are outlined in the sections that follow and are laid out in terms of short-, mid-, and long-term career objectives.

Short-term objectives (1-2 years):

Completion and submission of manuscript titled: Correlates of sexual functioning in lesbian women: A normative study.
Submission of an R21 grant application in response to PA-04-153: HEALTH DISPARITIES AMONG MINORITY AND UNDERSERVED WOMEN. The proposed project will be an educational intervention designed to increase lesbians' knowledge and awareness of risk factors for cervical cancer and to increase their participation rates in routine cervical cancer screening.
To complete data collection and analysis of data for a pilot project related to the evaluation of gender, race, and geographic disparities in tobacco-related co-morbidities and health behaviors among lung cancer patients. Data from this pilot project will be used in several planned manuscripts and will serve as preliminary data for a larger grant that will examine gender differences in post-diagnosis health behaviors and quality of life among cancer survivors.
Submission of manuscript(s) related to pilot project of gender, race, and geographic disparities in tobacco-related co-morbidities and health behaviors among lung cancer patients.
To submit grant applications in support of a research proposal to examine gender differences in post-diagnosis health behaviors and quality of life among lung cancer survivors.
Submission of first author manuscripts for completed projects (2-3 first author manuscripts per year).
Obtain additional formal and informal training in: 1) cancer epidemiology, prevention, and control; 2) designing, conducting, and evaluating longitudinal research data using advanced statistical methods in the area of cancer survivorship; and 3) clinical trials methodologies.

Mid-term objectives (3-5 years):

To be fully transitioned to independent (non-MORE-WH) research funding by the start of year 3.
To complete an exploratory study of an educational intervention to increase cervical cancer screening rates among lesbians.
Submission of first author manuscripts for completed projects (2-3 first author manuscripts per year).

Long-term objectives (5-10 years):

To be competively ready for promotion to the rank of Associate Professor by year 6.
To have on-going grant support for subsequent projects related to her two research focus areas: 1) cancer prevention among under-served women; and 2) gender differences in health behaviors and quality of life among cancer survivors.

IV. Specific Research Interests in Women's Health

During the latter part of her graduate training and during her first year as a junior faculty member in the Department of Epidemiology and Preventive Medicine, Dr. Tracy worked almost exclusively as a provider of informatics and statistical support to senior investigators. She has collaborated on several projects that have included a gender-based focus (e.g. outcome following traumatic injury, osteoporosis in men, interstitial cystitis) and has become increasingly interested in pursuing her own interests in women's health research.

Dr. Tracy's interest in women's health research is focused broadly on women and cancer. Under this broad umbrella, she is specifically interested in two primary focus areas: 1) cancer prevention in underserved women, particularly lesbians; and 2) health behaviors, quality of life, and psychosocial adjustment of female cancer survivors. Dr. Tracy has forged collaborations with Drs. Mark Krasna and Renee Royak-Schaler to support these research interests. In 2004, Dr. Tracy was awarded a small grant (Drs. Krasna and Royak-Schaler are collaborators) from the Maryland Cigarette Restitution Fund Program to examine gender, racial, and geographic differences in tobacco-related co-morbidities and health behaviors among lung cancer patients. Data collection for this project is currently in progress, and she hopes to use these data to develop a larger project to examine gender differences in survival and quality of life following lung cancer diagnosis.

Appointment to the MORE-WH scholars program will allow Dr. Tracy to focus significantly more of her academic energies to her own research agenda and to focus on two key aspects of her research program. First, it will allow her to devote larger amounts of time to completion of the data collection, statistical analysis, and publication of results for her pilot project related to gender, race, and geographic differences in tobacco-related co-morbidities and health behaviors among lung cancer patients. This is a project which directly relates to her research agenda focus on health behaviors, quality of life, and psychosocial adjustment of female cancer survivors and will provide valuable pilot data for larger grant proposals designed to explore these issues in larger samples and among various types of female cancer survivors. In addition, using pilot data obtained as part of her dissertation project, Dr. Tracy will be able to devote substantial effort to the development of a grant proposal in response to PA-04-153: HEALTH DISPARITIES AMONG MINORITY AND UNDERSERVED WOMEN. This proposal will be designed as an educational intervention study to increase lesbians' knowledge and awareness of risk factors for cervical cancer and to increase their participation rates in routine cervical cancer screening. This study will be a randomized controlled trial in which lesbians are assigned to one of three groups: 1) basic information about cervical cancer and recommended guidelines for screening provided at the baseline visit only (basic); 2) basic information at baseline and a follow-up reminder notice to get an annual pap exam (basic + reminder); and 3) basic information at baseline and a follow-up reminder notice to get an annual exam; group 3 follow-up reminder will also include information regarding lesbian-friendly clinics and clinics that provide pap screening for free or on a sliding scale fee schedule (basic+reminder+clinic information).

Gregory E. Hicks, M.P.T., Ph.D.
Appointment date: 7/1/2005 - 1/31/2007

Statement of Research Plan

I. Mentor Team

Jay Magaziner, Ph.D., M.S.Hyg., Professor, Department of Epidemiology & Preventive Medicine, School of Medicine (Lead Mentor)
Mona Baumgarten, Ph.D., Associate Professor, Department of Epidemiology & Preventive Medicine, School of Medicine
Patricia Langenberg, Ph.D., Professor, Department of Epidemiology & Preventive Medicine, School of Medicine
Alice Ryan, Ph.D., Associate Professor, Department of Medicine, School of Medicine.

II. Professional Goals

Dr. Hicks' professional goals were established early in his career as a physical therapist. He has been focused on improving the functional capacity and overall quality of life for all individuals with mobility limitations or disability. Early in his career, Dr. Hicks' methodology for achieving his goals was via individual patient interactions in clinical practice. As his interests have evolved through academic and research training, Dr. Hicks has learned to combine his clinical and epidemiological research skills so that his focus is more population-based. As evidenced from his research path, Dr. Hicks has also refined his focus to the exploration of mobility limitations in the geriatric population.

Ultimately, Dr. Hicks' long-term goal is to become a productive and independent investigator in the area of rehabilitation epidemiology with a focus on geriatrics and mobility disability. As a crucial part of this goal, Dr. Hicks would like to develop and evaluate interventions to improve the functional recovery pathway for older individuals following hip fracture, which is a particularly common disabling event for women. In order to meet this long-term goal, Dr. Hicks will need to devote more time to research activities so that he may acquire all of the necessary research skills to pursue new work in this area. Dr. Hicks believes that more structured mentoring during his current career phase will allow him to successfully transition from a junior investigator to a fully independent researcher who is capable of initiating, designing, and overseeing large-scale prospective studies.

III. Specific Research Interests in Women's Health

The research study that Dr. Hicks is proposing is based on his professional interests and builds on his current work and previous experience. The project represents his efforts to better understand specific factors associated with functional recovery from hip fracture in older women, thus informing the development of specific interventions for the hip fracture population. In the United States alone, more thant 350,000 people over the age of 65 are hospitalized every year for hip fractures; 80% of these among women. Current estimates suggest that by 2050, there will be 700,000 hip fractures annually. With this growning public health problem among older women, it is important to understand ways to prevent hip fracture and to successfully rehabilitate persons after hip fracture. At this point in time, very little is known about the role of rehabilitation in the recovery process from hip fracture or how rehabilitation may interact with interventions from other disciplines, such as medicine, nutrition, nursing, and social work.

Often, people who sustain a hip fracture do not attain a complete functional recovery following rehabilitation. Since there is an increased prevalence of fall risk factors following a hip fracture and the majority of hip fractures result from a fall, the high-risk hip fracture population deserves special attention to prevent falls and further fractures related to their mobility limitations. At this point, little attention has been given to the reduction of future falls in the vulnerable hip fracture population. Previous work in the area of fall prevention has largely focused on lower extremity training interventions, but Dr. Hicks' recent work suggests that trunk muscles also may play a critical role in maintaining balance in healthy older adults. Since poor balance has been consistently linked with mobility deficits and falls among older adults, efforts to improve and maintain mobility status should include attention to trunk muscle integrity, particularly in the hip fracture group.

Dr. Hicks' central hypothesis is that poor trunk muscle composition (higher levels of intramuscular fat infiltration and/or lower cross-sectional area) is associated with development of mobility disability and with a poorer recovery trajectory following an acute traumatic event in older women. The following specific aims were developed to test this hypothesis:

Specific aim 1: To determine whether poor trunk muscle composition is independently associated with falls in a healthy older population.
This aim will be achieved through secondary analyses of the Health ABC dataset, which has baseline CT data on trunk muscle composition and data on annual fall occurrences. Dr. Hicks' previous work in trunk muscle composition was done in this cohort. The Health ABC study is a prospective cohort study aimed to investigate the impact of changes in body composition and health conditions on age-related physiological and functional status. While this was a healthy, well-functioning and non-disabled population at the baseline, the older age of participants (between 70 and 79 years at the first clinic visit) makes them at risk of health-related events. The Health ABC population (n=3,075) is evenly distributed across gender and it is also characterized by a large number of African-American particpants (41.7%), so that it will be possible to explore whether gender and race differences influence disability-related mechanisms.

Specific aim 2: To determine whether poor trunk muscle composition is independently associated with poorer functional recovery in women following hip fracture.
This aim will be achieved as an ancillary study to the parent study entitled: "Sequelae of Hip Fracture in Men" (PI: Jay Magaziner, PhD), where most of the necessary measures will be obtained with the exception of CT scans. The parent study is designed to examine factors assoicated with functional recovery after hip fracture in a cohort of 200 men and 200 women over a 2-year period. Specifically, Dr. Hicks would like to have CT scans of the trunk muscles done on a subset of the cohort (100 women) at two months post fracture and at the one year follow-up. This will allow him to look at initial trunck muscle composition and change in trunk muscle composition as predictors of functional recovery. Functional recovery will be measured as an improvement in performance-based measures of physical function, i.e. walking speed and chair rise time.

Specific aim 3: To execute a pilot clinical study focused on specific trunk muscle exercise in older women following hip fracture with the following goals:

To examine the safety, feasibility and effectiveness of a combined trunk stabilization and lower extremity strengthening program versus a lower extremity strengthening program alone in improving physical function, reducing fall risk factors and improving balance confidence, as measured by a falls self-efficacy scale.
To obtain preliminary data on body composition and physical performance outcomes for use in design of a large scale, randomized, controlled study.

Laundette P. Jones, Ph.D.
Appointment date: 7/1/2005

Statement of Research Plan

I. Mentor Team

Angela Brodie, Ph.D., Professor, Department of Pharmacology & Experimental Therapeutics, School of Medicine (Lead Mentor)
Jodi Flaws, Ph.D., Associate Professor, Department of Epidemiology & Preventive Medicine, School of Medicine
Amy Fulton, Ph.D.,Professor, Program in Oncology and Department of Pathology, School of Medicine
Larry Anderson, M.D., Professor, Department of Anatomy & Neurobiology, School of Medicine

II. Background: The Role of Environmental Toxins in BRCA1-induced Mammary Cancer
BRCA1 is a breast cancer susceptibility gene whose role has been linked to a broad number of cellular processes including DNA repair, cell cycle checkpoint control, protein ubiquitylation, chromatin remodeling, transcriptional regulation, and most recently, protection against oxidative stress. Approximately 80% of hereditary breast cancer cases are caused by mutations in the BRCA1 gene. Although BRCA1 mutations are rare in sporadic cancers, several lines of evidence indicate that loss of BRCA1 resulting from reduced expression or incorrect subcellular localization may be important in non-familial breast cancer caused by other mechanisms, such as CpG methylation. Therefore, the discovery of the underlying mechanisms behind BRCA1-related breast cancers will have important implications that are relevant for both hereditary and sporadic breast cancers. Furthermore, this research could lead to the development of new prevention and screening strategies that target the evolutionary process of breast carcinogenesis.

The estimates for lifetime breast cancer risks among carriers of mutations in the BRCA1 gene range from 50% to 73% by age 50 and 65% to 87% by age 70. Therefore, ~20-50% of carriers live to advanced ages without developing the disease, which suggest that other factors may modify carriers' risks, including environmental exposures. Dr. Jones and her laboratory previously reported that tamoxifen stimulates mammary epithelial cell proliferation and ductal growth, and promotes the development of mammary adenocarcinomas in a mouse model with loss of full-length BRCA1. Additionally, their preliminary data showed that loss of BRCA1 was associated with enhanced ductal elongation during puberty. They also demonstrated in the post-pubertal gland that exogenous 17²-estradiol (E2) exposure resulted in sustained mammary epithelial cell proliferation, atypical ductal growth and the development of foci of aberrantly proliferating Estrogen Receptor ± (ER±) negative mammary epithelial cells. These results support the hypothesis that exposure to estrogen and compounds exerting estrogen-like effects can trigger an abnormal growth reponse in BRCA1-deficient mammary epithelial cells. Therefore, Dr. Jones will investigate the hypothesis that toxins that are classified as environmental estrogens will promote the development of BRCA1 mutation-related breast cancer by increasing estrogen growth stimulation using the chemical Bisphenol A as a model for environment estrogen exposure.

III. Hypothesis
Toxins that are classified as environmental estrogens will promote the development of BRCA1 mutation related breast cancer by increasing estrogen growth stimulation.

Specific Aims:

Determine whether exposure to Bisphenol A during prepuberty or puberty alters mammary gland development.
Determine whether exposure to Bisphenol A influences the risk of BRCA1-initiated preneoplasia and tumorigenesis.
Determine whether exposure to Bisphenol A during preinatal development alters mammary gland development and/or the risk of BRCA1-initiated preneoplasia and tumorigenesis in female offspring.

IV. Rationale
There are many compounds in the environment reported to have estrogenic activity. For example, Bisphenol A (BPA) is a building block widely used to make polycarbonate plastic products used in dentistry and the food packaging industry. It can mimic estrogenous E2 in vitro and in vivo through binding the ER and modulating target gene expression. Dr. Jones chose this chemical to study first because it has been widely investigated and it follows a biologically plausible mechanistic pathway to tumorigenesis. Additionally, recent findings have shown that in utero exposure of rodents to low, environmentally relevant doses of BPA induces precocious puberty, disruption of estrous cyclicity, and developmental changes in the mammary glands.

Research Design
A genetically defined mouse model that develops mammary adenocarcionomas with similar genetic changes to those found in human BRCA1 mutation-related cancers will be compared to non-transgenic control mice using a combination of established in vivo and in vitro techniques. BPA will be administered orally at either a low or high, environmentally relevant dose of 25 and 250 mg/kg, respectively. Mice exposed to E2 will serve as positive controls and mice exposed to ICI (a selective inhibitor of the estrogen receptor) or ovariectimized mice will serve as negative controls. In mice with intact ovaries that have undergone puberty, vaginal smears will be assessed regularly to determine estrous cyclicity.
For Specific Aim 1: To determine whether exposure to Bisphenol A during prepuperty (10 days old) or puberty (1.5 months old) alters mammary gland development, the following endpoints will be examined: (a) changes in the numbers of terminal end buds and extent of ductal development by whole mount analysis, (b) the percentage of tissue occupied by ducts, terminal end buds, terminal ducts, alveolar structures, and presence of secretory products by analysis of H&E stained histological sectons, and (c) changes in rates of cell proliferation and cell survival apoptotic indices by immunohistochemical (IHC) analysis.
For Specific Aim 2: To determine whether exposure to Bisphenol A influences the risk of BRCA1-initiated preneoplasia in vivo, the following endpoints will be examined in mice at 2, 4, and 6 months of age: (a) mammary hyperplasia and hyperplastic alveolar nodules (HANs) will be identified on H&E stained histological sections and whole mount analyses, respectively, and (b) changes in rates of cell proliferation and cell survival apoptotic indices will be determined by IHC. As a parallel approach, mammary gland organ culture in vitro studies will also be performed to examine the effect of Bisphenol A on BRCA1-initiated preneoplasia by using the established "Induction of nodule-like-alveolar assay." If a HAN is detected, then as a sub-aim, Dr. Jones will transplant the HAN into nude mice to determine whether tumors develop. To determine whether exposure to Bisphenol A influences the risk of BRCA1-initiated tumorigenesis, mice will be monitored at weekly intervals up to 12 months of age for tumor development. Tumor development will be recorded as the animal age when the tumor is first detected by palpation. Complete necropsies will be performed to examine for the presence or absence of non-palpable tumors and metastases. Estrogen and progesterone receptor expression will be evaluated in both in vivo and in vitro studies by RT-PCR and immunohistochemistry.
For Specific Aim 3: To determine whether exposure to Bisphenol A during perinatal development alters mammary gland development, female offspring will be examined at 10 days, 1.5 and 6 months of age to record changes in the endpoints specified in Aim 1. Exposure to BPA will begin on day 9 of pregnancy (the beginning of organogenesis in fetus). As a sub-aim, Dr. Jones will determine whether exposure to Bisphenol A during perinatal development alters the risk of BRCA1-initiated preneoplasia and tumorigenesis in female offspring using the endpoints outlined in Aim 2.

V. Limitations, Alternative Strategies and Future Directions
Dr. Jones does not anticipate significant problems with execution of the experiments proposed in this study because the molecular, histological, in vivo and organ culture techniques already have been established. Since she will be using mice with a C57B1/6 background and the Bisphenol A concentrations proposed in this study are based on published work by Markey and co-workers using a CD-1 mouse strain, the doses are not appropriate. In this case, a dose-response analysis of Bisphenol A will be performed for her mouse model. One limitation is that studies have shown that exposure to Bisphenol A can also induce tissue oxidative stress. In light of recent data by Bae et al., that suggest that BRCA1 may have a role in protection against oxidative stress, as a future direction Dr. Jones will investigate whether there are any modifications in endogenous antioxidant capacity, including superoxide dismutase (SOD), glutathione peroxidase (GPx), and catalase as a result of Bisphenol A exposure. Another limitation is that the carcinogen induced HANs in sub-aim III may fail to form tumors after transplantation and instead simply develop ductal or ductal-aveolar outgrowths. In this case, Dr. Jones would investigate whether multiple exposures to BPA are necessary for tumor development. If Bisphenol A does promote preneoplasia and tumorigenesis, then as a future direction, Dr. Jones will test whether estrogen-induced growth stimulation is the only mechanism contributing to this disease or if other pathways including genetic instability, oxidative damage or stimulation of other growth factor pathways also contribute. In future studies, Dr. Jones plans to test other environmental estrogens in this proposed model system, including DDE, dieldrin, and methoxychlor, to determine if any of these compounds influence the risk of BRCA1-initiated preneoplasia and tumorigenesis.

Tami J. Kingsbury, Ph.D.
Appointment date: 1/1/2006

Statement of Research Plan

I. Mentor Team

Angela Brodie, Ph.D., Professor, Department of Pharmacology & Experimental Therapeutics, School of Medicine
Amy Fulton, Ph.D., Professor, Program in Oncology and Department of Pathology, School of Medicine
Margaret McCarthy, Ph.D., Professor, Department of Physiology, School of Medicine

II. Background
Dr. Kingsbury's interest in calcineurin began when she was a postdoctoral fellow in the laboratory of Dr. Kyle Cunningham at Johns Hopkins University. Utilizing genetics and molecular biology to investigate calcium signaling in the budding yeast Saccharomyces cerivisiae, she discovered a novel family of proteins conserved from yeast to humans that function to regulate calcineurin activity in vitro and in vivo. One of the human family members, DSCR1, is encoded on chromosome 21 within the Down Syndrome Critical Region. Since calcineurin signaling is a crucial component of numerous signaling cascades and developmental processes, Dr. Kingsbury became interested in the possibility that inhibition of calcineuirn due to DSCR1 overexpression could contribute to some of the phenotypes associated with Down Syndrome. In order to pursue her interest in DSCR1 and Down Syndrome, Dr. Kingsbury decided to move from yeast into the mammalian system. She joined the laboratory of Dr. Bruce Krueger in the Department of Physiology at University of Maryland, Baltimore because of Dr. Krueger's expertise in developmental neurobiology and experience working with mouse models of Down Syndrome. Dr. Krueger's laboratory had previously demonstrated that calcium signaling directly regulates trkB expression in primary mouse cortical neurons. More recently, Dr. Kingsbury has found that calcineurin activity is required for calcium-stimulated trkB expression. Thus, DSCR1-dependent reduction in calcineurin function may underlie some of the cognitive deficits associated with Down Syndrome by impairing BDNF/trkB signaling. Dr. Kingsbury is currently seeking to elucidate the mechanism by which calcineurin modulates trkB gene expression. Since estrogen is known to reduce calcineurin function in neurons, she is also interested in the possibility that estrogen-mediated alterations in calcineurin activity may be involved in gender differences observed in the developing brain. Although BDNF signaling is typically thought of in reference to neuronal survival and function, trkB expression is also associated with aggressive tumors and poor patients prognosis in various forms of cancer such as lung lymphomas and prostate cancer. Recently, overexpression of trkB has been shown to prevent anoikis, suggesting a role for trkB in promoting the survival of metastatic cancer cells. In collaboration with Dr. Stuart Martin (University of Maryland Greenebaum Cancer Center), Dr. Kingsbury is screening a panel of breast cancer cell lines with varying metastatic potential for trkB expression. She hopes to use breast cancer cells to explore the relationship between DSCR1, calcineurin and trkB. This research is particularly relevant now that inhibition of calcineurin by DSCR1 overexpression has been proposed to mediate the protective effect of Down Syndrome against certain types of cancer, including breast cancer.

III. Professional Goals
Dr. Kingsbury's career goal is to establish an independent research laboratory at an institution where she can conduct research and continue teaching. She is particularly interested in a medical school setting where she can more readily develop collaborations with both basic researchers and clinical investigators. Although her laboratory will be primarily focused on understanding molecular details of biological processes, she hopes to develop strong collaborations with researchers using animal models in order to translate findings obtained in her laboratory to the whole organism.

IV. Research Interests
Down Syndrome (DS) occurs at a frequency of approximately 1 in 800 births. In addition to the distinctive facial features and mental retardation observed in Down Syndrome, it is now well documented that DS patients also exhibit a reduced incidence of solid tumors when compared to age-matched controls. The most striking protection is observed for breast cancer, which is extremely rare in DS patients. Although mental retardation and prevention of breast cancer may seem unrelated, available data suggest that inhibition of calcineurin, resulting from the DSCR1 overexpression, is a common molecular mechanism in the development of these two distinct phenotypes. Dr. Kingsbury is interested in investigating the role of DSCR1 and calcineurin in breast cancer development. This research will also address the potential of calcineurin regulatory proteins to be used as therapeutic targets for cancer intervention.

Dr. Kingsbury has chosen to use breast cancer cell lines to test this hypothesis for several reasons. First, since breast cancer is the form of cancer most dramatically reduced in DS patients, one would predict that development of breast cancer is particularly sensitive to calcineurin function and therefore provides an ideal assay system to investigate the regulation of calcineurin by DSCR1. Second, since calcineurin can be regulated by estrogen, the availability of both estrogen responsive and unresponsive breast cancer cell lines provides a unique opportunity to probe the effects of this critical hormone on calcineurin function. Breast cancer cell lines are also an ideal system in which to study how DSCR1 regulates calcineurin function. In contrast to primary neurons, breast cancer cell lines offer the advantage of generating sustainable cell lines with relatively rapid cell cycles that are amenable to molecular and biochemical assays. In addition, breast cancer cell lines provide an opportunity to conduct screens in order to identify novel functional protein interactions, as Dr. Kingsbury has done with yeast.

Specific Aim 1: The goal of this aim is to assess the role of DSCR1 and calcineurin in breast cancer. The effects of inhibiting calcineurin function by overexpression of DSCR1 or the application of the immunosuppressants FK506 or cyclosporine A on cyclin D1 expression, cell proliferation and resistance to anoikis will be assayed using available breast cancer cell lines. Once the role of calcineurin has been defined, assays can be developed to allow the regulation of calcineurin function to be investigated.

Specific Aim 2: Estrogen has been shown to decrease calcineurin activity in hippocampal neurons. In this aim, Dr. Kingsbury will test the effects of estrogen on calcineurin activity in breast cancer cell lines. Inhibition of calcineurin by estrogen may contribute to the gender differences observed in the cancer susceptibility of DS patients (females exhibit greater protection than males). Understanding the mechanism by which estrogen regulates calcineurin activity could be a significant portion of a future R01 application, with broad implications beyond the scope of breast cancer biology.

Future Aim: Once she has established herself as a breast cancer researcher, Dr. Kingsbury would like to expand the research in her laboratory to include the potential role of trkB in the development of metastatic breast cancer. The recent implication of DSCR1 in the protection of DS patients from cancer, trkB in the prevention of anoikis, and Dr. Kingsbury's findings that DSCR1 may protect against breast cancer at two levels: 1) Preventing solid tumor formation by restricting calcineurin/NFAT signaling or calcineurin/TORC signaling, and 2) decreasing metastatic potential by inhibitng inappropriate expression of potential oncogenes such as trkB.

Sharon M. Gordon, D.D.S., M.P.H., Ph.D.
Appointment date: 1/1/2006

Statement of Research Plan

I. Mentor Team

Joel D. Greenspan, Ph.D., Professor, Department of Biomedical Sciences, Dental School
Patricia Langenberg, Ph.D., Professor, Department of Epidemiology & Preventive Medicine, School of Medicine
Braxton D. Mitchell, Jr, Ph.D., Professor, Departments of Medicine and Epidemiology & Preventive Medicine, School of Medicine

II. Introduction
Temporomandibular disorders (TMD) is a condition accompanied by pain in the TMJ and associated muscles and is associated with depression, headache and back pain, significant use of health services, increased use of sedative and hypnotic medications, and increased risk for treatment-related injury, which may exacerbate the condition. The basis for TMD pain is not well understood; however, a notable feature of patients with painful conditions of the muscles and joints, including TMD, is that they occur more frequently in women than men, occurring at twice the rate, and affecting approximately 11 million people in the U.S., the majority of them women. Gender differences in pain response may be due, in part, to sex hormones and genetic factors. For example, TMD is more common among women in their reproductive years and decreases with increasing age. Hormonal replacement with estrogen or the use of oral contraceptives can worsen TMD. Studies in mice suggest an estrogen-mediated pain control system as well as differences in response to pain relieving drugs.

Importantly, sex differences in pain mechanisms and pain sensitivity have also been demonstrated in clinical studies of experimental pain, indicating that variation in gene expression is gender-dependent. Taken together, these studies sugggest a relationship between being female (the female "genotype") and factors related to chronic pain.

The mechanisms of gender-related pain sensitivity are only beginning to be identified. The majority of evidence available is taken from animal models and from experimentally-induced acute pain studies in healthy volunteers that may not accurately reflect the processes occurring in patients with chronic pain, such as TMD pain. The lack of understanding of this condition has resulted in a wide variety of therapies for the management of TMD, each based on a different perceived cause of pain and dysfunction. These treatments may include dental therapy (filing down teeth, placing caps on teeth, bite splints), myotherapy (trigger point injections, physical therapy), surgery (arthrocentesis, disc realignment, joint replacement with implants), medications, and behavioral therapy. There is no clear agreement, however, within the dental or medical professions on what constitutes appropriate treatment, partly due to the lack of clinical studies for this condition. NIH held a Technology Assessment Conference on the Management of Temporomandibular Disorders (1997), which recommended research on contribution of gender to TMD, as well as exploration of other " . . . risk factors, using predictive and explanatory approaches."

Lack of research has resulted in a paucity of evidence-based treatment approaches. This is exemplified by the treatment of TMD patients with proplast implants. In a study of TMJ implant patients with retained and removed implants, patients had pain that persisted long after removal of their failed implants. Pain severity increased with increasing frequency of surgeries, with nearly 80% of patients who reported severe pain having overall bodily pain meeting the ACR criteria for diagnosis of fibromyalgia (Ta 2002). These findings suggest that TMJ surgery, or the implants themselves, impart sufficient tissue injury to lead to sensitization resulting in hyperalgesia manifesting as greater clinical pain and additional painful conditions, such as fibromyalgia.

The available evidence suggests a need to study the relationship between genotype, hormonal influences, neurohumoral factors, tissue injury and associated mechanisms, and chronic pain. While it would be impossible to assess the many complex factors involved in TMD, with current technology it is possible to examine differences in one or more prototypic markers for hormonal activation, inflammation, and/or connective tissue adaptation/dysfunction, and genotype.

III. Objectives
The study objective is to investigate factors related to persistent pain in TMD patients, with the long-term goal of identifying targets for pain prevention and enhanced analgesic response.

IV. Questions, Hypotheses, and Specific Aims
Data from the work of Dr. Gordon and others has resulted in unresolved questions of whether tissue injury from treatment of TMD leads to sensitization resulting in hyperalgesia manifesting as persistent and disseminated pain. Moveover, what are susceptibility factors leading to manifestation of persistent pain in some patients versus others? To address these questions, Dr. Gordon will conduct a case control analysis of TMJ implant registry data with the following aims:

Evaluate the relative contribution of TMD and surgical treatments on manifestation and maintenance of persistent pain.
Hypothesis: Patients receiving surgically-based treatment are more likely to have painful co-morbidities, such as fibromyalgia, CRPS, or other pain syndromes.
Evaluate the genetic contribution to pain sensitivity in TMD patients.
Hypothesis: There is an association between genetic polymorphisms identified as related to pain susceptibility and patients experiencing persistent pain, specifically examining: mu and delta opioid, COMT, and alpha-adrenergic receptor genes.

Dr. Gordon's hypotheses are based on the literature that indicates:

Persistent pain is due to neuronal plasticity.
Tissue injury and subsequent inflammation contribute to plasticity.
Pain is highly variable across patients.
Genetic variability in pain and analgesic response has been demonstrated in animal studies and in clinical experimental pain.
Sex differences in pain mechanisms and pain sensitivity have been documented and appear related to genetic factors.

Of the many possible genetic polymorphisms studied in rodent models, Dr. Gordon has chosen to study those that have been shown to be assoicated with experimental pain in humans: the mu and delta opioid, COMT, and the alpha-adrenergic receptor gene.

V. Data Source and Analysis Plan
The data source is the NIDCR TMJ Implant Registry (TIRR), which has clinical data and banked specimens for patients with TMD who have not had surgical treatment (n = 159), TMD patients having had a non-implant surgical treatment (n = 149), and TMD patients who have had implants placed or placed and removed (n = 158). TMD patients without surgery will serve as the reference group. All registrants have been classified according to the TMD Research Diagnostic Criteria. The diagnosis, as well as medical diagnoses, surgical history, and pain report are available clinical data. In addition, stored specimens include blood and saliva stored for DNA analysis as well as separated fragments (serum and saliva supernatant).

For the outcome of persistent pain, factors of interest include surgical history and number of and types of surgeries as indicators of tissue injury, medical diagnoses and RDC diagnosis, cytokines indicative of an inflammatory response, and genotype. Pain at entry into the registry will be an adjustment factor. Potential contributing variables to be examined include:

Age, sex and other demographic variables
Insurance and marital status
Time since diagnosis
Precipitating events (if any)
Medications
Co-morbidities (medical diagnoses)
Psychological factors
Head and neck examination findings, including radiographic

Using a 1:4 ratio of cases to controls, Dr. Gordon will examine the association of persistent pain with the genetic polymorphisms indicative of a "pain sensitivity" genotype, as well as pro-inflammatory cytokines related to the responses to tissue injury and pain response.

Didactic Coursework

UMB offers a broad variety of short- and long-term programs intended to develop and improve skills in research design and conduct, statistical analysis, and specialized laboratory and other technical skills. MORE-WH scholars will develop an individualized didactic training curriculum with their mentor teams as part of the overall training plan based on an assessment of need and previous training and are encouraged to take advantage of the many courses offered on campus.

The following courses are suggested for MORE-WH scholars. For a complete list of courses, seminars and journal clubs offered by the Department of Epidemiology & Preventive Medicine Academic Program, go to Department of Epidemiology & Preventive Medicine Academic Programs and Training Opportunities. Other courses offered by the UMB Graduate School can be found at: http://graduate.umaryland.edu/

Introduction to Clinical Research
Biostatistics for Clinical Research (short course)
Responsible Conduct of Research
IRB Basic Training for Research: "Protecting Human Research Subjects"
Master of Science Degree in Epidemiology and Preventive Medicine: Clinical Research Track

Workshops & Seminars

Training in the Responsible Conduct of Research
Health Sciences and Human Services Library (HS/HSL) Classes: The HS/HSL offers UMB faculty, staff and students training in Microsoft Officer, database searching, bibliographic management, and web publishing. Class descriptions, dates and times can be found at: http://www.hshsl.umaryland.edu/.
UMB Graduate School Seminar Series
Women's Health Research Seminar Series
School of Medicine Dean's Office Student and Faculty Development Professional Development Series
Other Women's Health Conferences

Funding Opportunities