Congratulations to our 2008 Pilot Project Awardees!
Kent Christopherson II, PhD
Hazel Lum, PhD
Ece Mutlu, MD
Aloka Patel, MD
JoEllen Wilbur, PhD, RN
Qiping Zheng, PhD
Title: Regulatory Mechanisms Governing Blood Formation from Embryonic Stem Cells
PI: Kent Christopherson II, PhD Assistant Professor of Medicine and Immunology/Microbiology, Sections of Hematology and Stem Cell Transplantation
Abstract: Long lasting severe thrombocytopenia (decreased platelets) and neutropenia (decreased neutrophils) is observed in adult patients following receipt of umbilical cord blood cells as a donor source for Hematopoietic Stem Cell Transplantation. This situation results in an increased risk for serious bleeding, infection, and the risks associated with repeated blood transfusions. No clinical treatment strategies currently exist that are adequately effective at increasing the numbers hematopoietic stem cells or myeloid progenitor precursors of neutrophils or megakaryocytes in this clinical setting. A better understanding of the overall regulation of early blood development is therefore needed in order to more effectively suggest biological targets at which to intervene with novel treatment or cell therapy based strategies. This research project will evaluate the regulatory mechanisms involved in blood formation during embryonic stem cell differentiation. The clinical implications of this project are extremely high for future stem cell transplant patients as the project is anticipated to lead to new therapeutic strategies to manipulate the blood development for the benefit of patients.
Title: Pathogenic Role of Lysophosphatidylcholine, a PLA2-Derived Lipid, in Asthma
PI: Hazel Lum, PhD Professor, Department of Pharmacology
Co-PI: Mark Yoder, MD, Assistant Professor, Department of Medicine, Division of Pulmonary and Clinical Care Medicine
Abstract: The following summarizes the significance and major points addressed by the project: Asthma is a chronic disease of the lung airways correlated with T-helper-2 (Th2) inflammation and tissue remodeling, and as the disease progresses to the severe form, there is increased dominance by neutrophils, heightened tissue destruction and airway wall remodeling typical of a chronic wound-repair process. It is also evident that the more severe forms show reduced responsiveness to anti-inflammatory drugs, and inflammatory injury remains despite treatment with high-dose therapy, leading to unacceptable morbidity and mortality. To-date, the underlying factors that initiate and sustain the progression of asthma severity remain elusive, and there is a clear need of a more comprehensive understanding of the basis by which asthma progresses and persists.
Evolving evidence suggests that asthmatic patients and animal models of asthma may have high and chronic amounts of a known broad-spectrum pro-inflammatory lipid, lysophosphatidylcholine (LPC), since the key enzyme for its production, phospholipase A2 (PLA2) is elevated in bronchoalveolar lavage fluid (BALF), airway sputum, inflammatory cell infiltrates, and cells of the airway wall. We suspect that high chronic levels of LPC not only would injure airway epithelium and mobilize inflammatory cell infiltrates into the lungs, but also could contribute to progression towards the more severe form of asthma. Based on our prior studies of LPC in the vascular endothelium, we also know that LPC works through a novel lipid- and pH-sensing G protein-coupled receptor, GPR4, and our preliminary findings show that lung airway epithelial cells strongly express GPR4. Therefore, we propose two aims to investigate the working hypothesis that elevated levels of LPC are produced in asthmatic patients which induce airway epithelial injury and mobilize inflammatory leukocytes in a GPR4-dependent manner. In Aim 1, asthmatic patients will be recruited to obtain BALF and sputum samples to determine the levels of LPC by tandem mass spectrometry as well as PLA2 forms. In aim 2, the extracted samples of LPC from BALF and sputum from patients along with purified LPC as control will be evaluated for its ability to injure human bronchial and bronchiolar epithelium and to recruit inflammatory leukocytes using in vitro culture assays. The goal is to obtain biochemical and functional evidence to causally link LPC in asthmatic patients to the inflammatory airway epithelial injury, and thereby, providing the "proof-of-concept" for the hypothesis. For this work, we have a team of clinical and basic scientists in the lung field, providing a complementary effort for the investigation and for training of young scientists. If our findings hold true, the results will provide important novel insights into pathophysiologic mechanisms of asthma not recognized before, and could be the basis for future larger grant mechanisms.
Title: Development of a Molecular Method to Study Fungal Microbiota in IBD
PI: Ece A. Mutlu, MD, MBA, Director, IBD Program, Director, Clinical Research, Associate Professor of Medicine, Section of Gastroenterology and Nutrition
Abstract: Inflammatory Bowel Diseases (IBDs), namely ulcerative colitis (UC) and Crohn's disease (CD) are chronic, lifelong, relapsing illnesses, affecting close to 1 million Americans and costing approximately $2 billion/year to the US healthcare system. IBDs are of unknown cause, have no cure and are increasing in incidence. It is now widely accepted that IBD flare-ups are triggered by the many microbial organisms that live within the intestines. These are called the gut microbiota and are accepted to be the major antigens against which the immune system reacts in IBD. Despite the many clinical and experimental clues that an altered microbial composition may exist in IBD, the specific changes in the microbiota of IBD patients are largely unknown. Narrow searches for a single pathogen as the cause of IBD have been unsuccessful. A limited number of studies looking at multiple bacterial groups have shown ambiguous results in the composition of microbiota in IBD. On the other hand, methods to study large sets of organisms simultaneously have been lacking until now. Hence, the GI tract microbiota has even been termed the "neglected organ" in the body. Current studies are underway studying the bacterial gut microbiota in IBD by the PI as well as other researchers in the country; however, there are no studies that have tried to simultaneously examine the fungal microbiota. In fact, there are no reports of what constitutes a normal fungal gut flora using molecular biology techniques. On the other hand, it is also a well known fact that CD is characterized by the presence of anti fungal antibodies to Sacchromyces cervicae and fungal mannans: In fact, such anti-fungal antibodies have recently been linked to the severity of IBD in pediatric population. Furthermore, intestinal epithelial and immune cells carry specific receptors like toll like receptor 2 (TLR2); dectin receptors, mannose receptors, DC-SIGN receptors that are activated by fungal organisms, & some have been reported to be upregulated in IBD. A probiotic yeast (Sacchromyces boulardii) has been shown to keep CD in remission, pointing out the importance of studying the fungal GI tract microbiota in IBD. Thus, in this application, we propose to develop a fingerprinting assay to study of the GI tract fungal flora in IBD cases and healthy controls.
Title: The Impact of Early Enteral Feed Type on Initial Gut Microflora and Disease in Very Low Birth Weight Premature Infants
Title: P63 in Bone Development and Cancer Formation
PI: Aloka L. Patel MD, Assistant Professor, Pediatrics
CO-PI: Ece A. Mutlu, MD, MBA, Assistant Professor, Internal Medicine
Abstract: Infants who are born with birth weights below 1500g, also known as very low birth weight (VLBW) infants, are susceptible to many morbidities that are increasingly linked to inflammatory processes that originate in the gastrointestinal tract or gut in the early post-birth period. At birth, the infant's gut is sterile and quickly becomes colonized with bacteria, termed the gut microflora. In term healthy infants, the gut microflora is influenced primarily by the mode of delivery and the type of feeding, with human milk-fed infants having a predominance of "normal" commensal organisms that down-regulate inflammation, and formula-fed infants having a predominance of potentially harmful pro-inflammatory bacteria. Colonization of the gut is disrupted in hospitalized VLBW infants, setting the stage for inflammatory processes that act locally and systemically, leading to short-and long-term morbidities. Human milk may significantly impact the emerging gut microflora in VLBW infants because it contains a variety of biologically active substances that directly down-regulate inflammation, and has properties of both pre- and probiotics. However, previous gut microflora studies in VLBW infants are limited by the use of culture-based microbial techniques in which only 10-30% of gut bacteria can be cultured, and lack of information about the dose and exposure period of human milk and formula feedings, especially during the critical exposure period when the introduction of feedings and the emergence of gut microflora overlap.
The aim of this pilot study is to determine the effects of human milk and formula feedings over the first 14 days of life on gut microflora and morbidities in VLBW infants. Demographic, feeding and clinical data and biologic samples will be collected from 60 VLBW infants and their mothers. Gut bacterial fingerprints will be identified from rectal and stool samples using Amplicon Length Heterogeneity (ALH), a 16S rDNA PCR-based technique. These ALH bacterial fingerprints consist of extracting DNA from a sample, using PCR to amplify DNA pieces (called amplicons) of different lengths that reflect each type of bacteria present in the sample. The specific types of bacteria will be identified using high throughput DNA sequencing, a technology that is less expensive and less time consuming than traditional cloning and sequencing. The differences in gut microflora based on feeding type will be correlated with incidence of morbidities, specifically late onset sepsis, pneumonia and necrotizing enterocolitis in human milk-fed versus formula-fed infants. Thus, this study may start to clarify any potentially therapeutic role of human milk in the colonization of gut microflora for VLBW infants and could lead to new clinical tools to predict at-risk infants and evaluate preventive therapies such as human milk and/or pre-and probiotics.
Title: Cognition and Physical Activity Community Elderly Study (CAPACES)
PI: JoEllen Wilbur, PhD, MSN, BSN, Associate Dean for Research and Professor, Department of Women's and Children's Health Nursing, College of Nursing
& Co-PIs: Lou Fogg, PhD, BS, Assistant Professor, College of Nursing & Departments of Behavioral Sciences and Psychiatry
Beth A. Staffileno, PhD, RN, Assistant Professor, Department of Adult Health, College of Nursing
Martha Clare Morris, ScD, MS, BS, Associate Professor, Department of Medicine & Assistant Provost of Community Research
Robert Wilson, PhD, MA, BA, Professor, Dept Neurological Sciences
David Marquez, PhD, MS, BS, Assistant Professor, Department of Kinesiology and Nutrition, University of Illinois at Chicago
Abstract: Decline in cognition associated with aging is a key indicator of for Alzheimer's Dementia, thus identifying interventions/strategies that maintain and enhance cognition through the middle and older years of life is of major public health concern. Alzheimer's Dementia rises exponentially from 3% among persons 65 to 74 years to almost 50% among those 85 years and older. No where is the aging population expected to grow faster than among Latinos, which currently represent 14.8% of the population. Importantly, there is evidence to suggest that some Latinos in the US may be unduly burdened by factors placing them at risk for cognitive decline including challenges with language, low acculturation to US norms, and higher rates of diabetes than the dominate culture. Further, a recent Cochrane report provides evidence that physical activities which improve cardio-respiratory fitness are beneficial for cognitive function in healthy older adults, but unfortunately rates of reported leisure time physical inactivity in the US are among the highest for Latinos. Overall, however, research to date on both cognition and physical activity has not included older Latinos or low numbers have restricted comparisons. We know little about the overall physical activity of Latinos and, when it is narrowly defined as leisure time, it may be underestimated and opportunities to enhance physical activity missed. Thus, the current study aims to: 1) explores gender and age differences in physical activity among older Latino men and women. In the proposed study physical activity will be measured by self-report and activity counts (accelerometers), and aerobic fitness will be indirectly assessed; and 2) explore relationships between older Latino's physical activity and cognition.
PI: Qiping Zheng, M.D., PhD, Assistant Professor, Department of Anatomy and Cell Biology
Co-PI: Rick Sumner, PhD, Professor and Chair, Dept. of Anatomy and Cell Biology
Steven Gitelis, MD, Professor and Chair, Department of Orthopedic Surgery
Guojun Wu, PhD, Assistant Professor, Dept. of Pathology, Wayne State University
Huilin Yang, MD, PhD, Professor and Chair, Dept. of Orthopedic Surgery, Soochow University, China
Abstract: Apoptosis is a form of programmed cell death that occurs during both normal tissue development and cancer formation. p53 family members, including p53, p63 and p73, are well-known tumor suppressors. p53/p63 stimulation of apoptosis is one of the major mechanisms that prevent tumor formation. Interestingly, both p53 and p63 have been linked to bone cancer formation and bone development. Type X collagen is a specific molecular marker of hypertrophic chondrocytes, an end stage of cartilage development during bone formation. As a matrix protein, type X collagen also functions as a regulatory molecule that affects chondrocyte mineralization, apoptosis, and angiogenesis. These processes are closely related to skeletal development and cancer formation. However, until recently, the specific factors that regulate type X collagen gene (Col10a1) expression in vivo have not been identified yet. We recently identified that p53 related proteins, especially, p63, might be candidate factor that contribute to the regulation of Col10a1 expression. We hypothesize that p63 dysregulation of Col10a1 may account for certain skeletal defects and bone cancer formation as well. These pathophysiological conditions may share common mechanisms that lead to delayed/accelerated chondrocyte maturation and pro- or anti-apopototic events and thus link skeletal development with bone cancers. By successful accomplishing this pilot project, we expect to further address the mechanic basis of p63 regulation of Col10a1 during bone development and cancer formation and eventually, lead to identification of candidate molecules that have diagnostic and therapeutic value preventing skeletal disease progression and bone cancer formation.