You are here: SURGERY HOME > RESEARCH > NIH FUNDING
NIH Funding
This mosaic was created by General Surgeon Dr. Harold Berg (1918-2002). Close to 200 of Dr. Bergs mosaics are on display in Louisville and around the world. A native of Brooklyn, NY, Dr. Berg graduated from the University of Louisville School of Medicine in 1941. He had an established general surgery practice and began teaching at the University of Louisville in 1961. Dr. Berg never accepted payment for any of his art works, which were always thoroughly researched and freely given out of respect to the recipients. Shown here is a mosaic of Dr. Abraham Flexner, who was born in Louisville (Ky) and is known for his 1910 publication titled the Flexner Report. An emphasis in the report was for increased faculties at university hospitals dedicated to teaching and research. Our dedication to this Louisville tradition is reflected in the research efforts of our PIs. Several of our faculty investigators have received awards from the National Institutes of Health. The research descriptions below are derived from the National Institutes of Health Research Portfolio Online Reporting Tool (RePORT), which is available at http://www.projectreporter.nih.gov/reporter.cfm.
PI Name: H. Sam Zhou
Title: Adenovirus E1B55K Functions Related to Oncolytic Replication
Grant Number: R01CA129975
Fiscal Year Search: 2010
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7777755&icde=5639745
DESCRIPTION (provided by applicant):
Two major "hallmarks" of all cancer cells include dysregulated cell cycle and inhibited apoptosis, both of which are also involved in adenovirus (Ad) infection. These two processes are primarily conducted by viral oncoproteins E1A and E1B. Since the function of viral E1 protein is similar to that of cancer cellular factors that promote proliferation and inhibit apoptosis, viruses with mutations in oncoprotein E1A and E1B can selectively replicate in cancer cells. One such mutant with deletion of E1B55K, known as dl1520, has been used in clinical trials documenting oncolytic effects. However, the mechanism of viral oncolytic replication has not been well characterized, and the therapeutic efficacy needs improvement. It is generally believed that the major role of E1B55K is to bind to and inhibit p53 activation, but many studies have documented that E1B55K-mediated p53 inactivation is not required for virus replication. Our laboratory has shown: (1) that the limited spread of mutant viruses in large tumors is a key factor in decreased therapeutic efficacy; (2) increased Ad E1A expression enhances virus-oncolytic replication; and (3) apoptosis caused by E1B deletion can partially decrease virus replication but does not change the final outcome of virus-mediated cancer killing. Our recent studies have revealed that E1B55K has a novel function in the induction of cyclin E and other cell cycle-related genes. Most importantly, we also observed that increased cyclin E expression is correlated with virus replication efficiency. E1B55K-induced cyclin E expression is required for virus replication in normal cells, but is not necessary in cancer cells. We hypothesize that E1B55K may target cellular factor(s) to increase cyclin E expression, and this factor(s) may already be activated in cancer cells. Thus, cyclin E dysregulation in cancer cells may be the molecular basis for oncolytic replication of E1B55K-deleted viruses. Our research team will (1) identify cellular factors targeted by E1B55K for cyclin E induction, (2) define the mechanism by which E1B55K activates cyclin E expression, and (3) determine the relationship between cyclin E overexpression and oncolytic replication of E1B-deleted viriuses. If we confirm that oncolytic replication relies on cyclin E expression and cell proliferation, patients with aggressively growing tumors and dysregulated cyclin E should greatly benefit from this adenoviral therapy. The long-term goal of this work is to increase the efficacy of oncolytic cancer gene therapy. PUBLIC HEALTH RELEVANCE: Adenoviruses lacking an important regulative protein-E1B55K-still can amplify in some cancer cells. Therefore, the E1B55K mutant dl1520 has been used in clinical trials. It is important to understand the E1B55K function and the selective replication of E1B55K-deleted dl1520 in cancer cells. Our recently published studies have revealed that E1B55K has a novel function in the induction of cyclin E expression, which is crucial for DNA replication. Cancer cells generally express high levels of cyclin E or have a dysregulation of the gene. We reason that viral E1B55K may activate some cellular factors that increase cyclin E expression for viral DNA replication. Cancer cells may already have the factors activated; therefore cancer cells do not require the E1B55K function. The study of this possibility is very important. If we confirm that mutated virus replication relies on cyclin E expression, patients with aggressively growing tumors and dysregulated cyclin E should greatly benefit from this adenoviral therapy. The long-term goal of this work is to increase the efficacy of oncolytic cancer gene therapy.
PI Name: Brian G. Harbrecht, MD
Title: Hepatocyte Nitric Oxide Synthase Regulation by Glucagon and Insulin
Grant Number: R01DK055664
Fiscal Year Search: 2010
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7743768&icde=5639745
DESCRIPTION (provided by applicant):
The immune response to sepsis involves a series of complex, highly integrated homeostatic responses that, if prolonged and excessive, can lead to organ dysfunction and death. Nitric oxide (NO) synthesis is upregulated by sepsis in many tissues and is an essential component of the host immune response. Nitric oxide synthesis can be beneficial and improve immune and organ function, but if synthesis is excessive and prolonged, NO can promote organ injury, tissue inflammation, and death. NO is produced in hepatocytes by the inducible nitric oxide synthase (iNOS) that is stimulated by cytokines and proinflammatory stimuli. Excessive NO from iNOS produces cellular dysfunction and hepatic injury. Glucagon and cyclic adenosine monophosphate (cAMP) regulate hepatic iNOS expression in vitro and in vivo, and by doing so, decrease NO-mediated hepatic injury. Our preliminary data demonstrate that insulin also down-regulates cytokine-induced iNOS expression. Both glucagon and insulin alter specific intracellular signaling pathways in hepatocytes, but the mechanisms involved in the regulation of hepatocyte function in sepsis by glucagon and insulin, and specifically the regulation of hepatocyte iNOS expression, have not been identified. In this proposal, we will determine the mechanisms responsible for the regulation of hepatocyte iNOS expression by glucagon and insulin. In Aim I, we will continue our work in determining the mechanism for the glucagon and cAMP-induced inhibition of hepatocyte iNOS expression. We will focus on protein kinase A (PKA)-independent pathways induced by cAMP and evaluate the role of the guanine nucleotide exchange factor Epac and the role of calcium. In Aim II, we will determine the mechanisms responsible for the inhibition of iNOS by insulin. By defining how these hormones regulate hepatocyte iNOS expression, we will provide a framework for understanding the basic pathophysiologic cellular events in shock and sepsis that may lead to novel cellular-based therapies for critically ill patients. Project Narrative: Nitric oxide is synthesized in critically ill patients during septic shock, and when overproduced, can increase cellular dysfunction, tissue injury, and death. Glucagon and insulin primarily regulate blood glucose, which has become an important facet of the care of critically ill patients, but we have found that they also regulate hepatic nitric oxide production. We will determine the mechanisms responsible for the regulation of hepatocyte inducible nitric oxide synthase (iNOS) expression by glucagon and insulin. By defining these mechanisms, we will provide a framework for understanding the basic cellular events in shock and sepsis, which may lead to novel cellular-based therapies for critically ill patients.
PI Name: Robert C.G. Martin
Title: Prevention of Reflux-Induced Esophageal Adenocarcinoma By Dietary Berries
Grant Number: R03CA137801
Fiscal Year Search: 2009
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7689145&icde=5640031
DESCRIPTION (provided by applicant):
Esophageal adenocarcinoma (EAC) has the fastest rate of increase among all cancers in America, yet there is very little research on the mechanisms and prevention of its development. EAC primarily affects White males and has one of the poorest prognoses among all cancers. Barrett's esophagus (BE) is a pre-cancerous lesion associated with the development of EAC. BE is caused by chronic heartburn, which affects more than 19 million Americans every year. Pharmaceutical treatments to cure heartburn do not appear to largely affect EAC incidence. New intervention strategies are urgently needed to address this issue. Commonly available berries, such as blueberries (BB) and black raspberries (BRB), are natural chemopreventative agents that can be easily incorporated into the American diet and used for clinical intervention. Moreover, dietary BRBs are highly effective in preventing squamous cell carcinoma (SCC), another type of esophageal cancer, the incidence of which is declining. We have developed an animal model that mimics the clinical development of EAC. In this model, chronic esophageal reflux resembling heartburn is induced in rats using a surgical procedure. More than one-third of the rats develop EAC at the end of 6 months. We can effectively use this model to study the preventive efficacy of dietary berries. Two types of berries have been chosen, since they differ widely in their phytochemical profiles. Our objective is to study the efficacy of dietary berries to prevent the development of reflux-induced EAC and to understand the mechanisms by which they do so. To achieve this, we plan to provide BB and BRB via the diet to male rats at a dose of 2.5%, which is equivalent of consuming one-half cup of dried berries every day. We will surgically-induce chronic reflux. At the end of 6 months, the rats that were provided with berry diet will be compared with those that did not receive intervention to evaluate the incidence of tumors. To further understand the mechanisms by which berries may prevent esophageal tumorigenesis, we plan to use intermediary monthly time points during the development of EAC to study the biomarkers that are altered. The first set of biomarkers deal with oxidative stress and will determine whether berries can induce cellular antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase, which ultimately protect the cells from oxidative damage. Lipid peroxidation and oxidative DNA damage will be used as markers of oxidative stress. The second set of biomarkers deal with inflammation associated with BE. Nuclear factor kappa B (NF?B), a transcription factor that controls the expression of several cancer-inducing genes, is found to be activated by inflammation in BE. We will evaluate whether berries can prevent this activation. Thus, by following a two-part approach, we will be able to determine whether berries prevent EAC progression and the mechanisms involved in this prevention.
PI Name: Sufan Chien, MD
Title: Intracellular Energy Delivery and Diabetic Wounds
Grant Number: R01DK074566
Fiscal Year Search: 2010
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7803637&icde=5639745
DESCRIPTION (provided by applicant):
The long-term goal of our program is to develop a safe and effective technique to combat various tissue ischemic damages. The specific aim of this proposal is to use our newly developed proprietary intracellular energy delivery technique to promote healing of diabetic wounds. Of the 17 million Americans with diabetes, approximately 2.5 to 4.5 million will develop a chronic wound in their lifetime. The overall cost of diabetic foot problems, including loss of productivity, could be as high as $20 billion per year. Despite thousands of dressing products developed to treat wounds, none have shown consistent effect. We propose a new approach for chronic wounds. Our central hypoth- esis is that wound tissue hypoxia results in depletion of adenosine triphosphate (ATP), which is the fundamental cause of non-healing chronic wounds, and a direct intracellular ATP delivery will improve microenvironment of wound tissue and facilitate healing process. Direct energy supply for wound treatment has never been attempted before, and the relationship between increased energy supply and wound healing process is entirely unknown. During the tenure of the Pi's NIH grant entitled "Enhanced glycolysis for hypothermic heart preservation", a new technique for direct intracellular delivery of ATP has been developed in which a special carrier is used to encapsulate ATP. The composition of this carrier is similar to the cell membrane. When the carrier meets with the cell membrane, it fuses with it and delivers the contents into the cytosol. Preliminary results indicate that this new energy delivery technique can provide significant protection to ischemic cells and tissues. The technique has shown very promising effects on normal and ischemic wounds. Three US patents and more than 12 international patents have been filed and the innovation has also been reported to the NIH. Our preliminary results also indicated that high-energy phos- phate contents were severely depleted in human chronic wounds, and treatment with ATP-vesicles in animal wounds increased tissue high-energy contents. Five hypotheses will be tested: (1) high-energy phosphate contents are decreased in chronic diabetic wounds; 2) an ischemic wound model created using a minimally invasive surgical technique can be tolerable to diabetic animals; 3) intracellular ATP delivery will increase wound tissue energy levels to facilitate healing; (4) by providing energy to wound tissue, improved healing is achieved through coordinated upregulation of growth factors and other healing mechanisms; and (5) direct intracellular energy delivery will enhance wound healing by improved tissue perfusion. These issues have not been explored in the past, but our preliminary results have established the basis for the success of this project. The expansion of usage of the direct intracellular energy delivery is likely to have a major impact on medicine. It will not only improve chronic wound care, but also help our treatment to various ischemic conditions, such as severe trauma, shock, stroke, heart attack, spinal cord injury, cardiopulmonary bypass, organ transplant, and many other acute and chronic ischemic diseases.
PI Name: James B. Hoying
Title: Fabricated Microvascular Networks
Grant Number: R01EB007556-05
Fiscal Year Search: 2010
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7822693&icde=5639745
DESCRIPTION (provided by applicant):
Fabricated Microvascular Networks. The importance of an effective vascular supply for tissue health is universally accepted. In developing strategies to build vasculatures for tissue engineering and other therapeutic applications, it is important to recognize that, foremost, the new vasculature must quickly provide sufficient blood flow to the target tissue to preserve cell viability. We have found that new microvessels formed in vitro can begin to carry blood within the first days following implantation. However, flow patterns are atypical and likely ineffective at establishing normoxia until many days later. The delay is primarily due to a lack of organization within the network at the time of implantation and the time needed to develop new mature inflow and outflow pathways. We hypothesize that pre-determining an appropriate network organization prior to implantation would reduce the amount of time needed for the new microvasculature to effectively perfuse a tissue. We have established generic technologies utilizing a direct-write tissue printing tool for patterning and organizing tissue components for tissue engineering applications. We propose to implement this technology to design and fabricate pre-patterned, 3-dimensional microvascular networks with pre-existing inflow and outflow pathways. Also, we will use an in vitro, intravascular-perfusion bioreactor system to establish flow through the networks to further organize and mature the microvascular networks prior to implantation. Computational modeling and physiological analyses serve to direct design strategies and characterize the architectures and functionality of the fabricated vasculatures both in vitro and in vivo. In addition to providing an enabling technology platform for assembling pre-determined microvascular networks, this work will provide a foundation from which to explore the importance of network architectures in vascular function.
PI Name: Suzanne T. Ildstad
Title: Training Program in Transplantation
Grant Number: T32HL076138
Fiscal Year Search: 2010
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7893659&icde=5639745
DESCRIPTION (provided by applicant):
This is a resubmission of a competing renewal for a highly successful training program in transplantation immunology and stem cell biology. Over the past 11 years, a dynamic group of internationally recognized investigators in transplantation and stem cell biology were strategically recruited to the University of Louisville which is increasingly recognized for its comprehensive translational research in composite tissue allotransplantation, tolerance, and stem cell-mediated regenerative medicine. This training program directly addresses the mission of the NIH training grant program to help ensure that a diverse and highly trained workforce is available to assume leadership roles related to the nation's biomedical research agenda. All positions have been filled with outstanding applicants to date including six women (one minority), thereby addressing the major concern of all three reviewers. Applicants must have a Ph.D., D.V.M., or M.D. They are required to submit a CV, publication list, three letters of recommendation, and a statement of the specific research proposed. An Executive Training Committee comprised of the program Director and senior trainers selects the trainees. Trainees assemble a program and undertake basic research with one trainer and one or more co-mentors. Evaluation of the Training Program occurs on several levels, including manuscript and abstract submissions; oral presentations and participation at research seminar series; and completion and submission of an NRSA postdoctoral individual training grant or equivalent (AHA, JDRF, ASH, or other foundation or society fellowships) within the second year of training. The overall progress of this Training Program is assessed by an External Advisory Board through semiannual meetings and feedback. The External Advisory Board also assists in minority recruitment. The faculty in this training program has the expertise and breadth of background to successfully train fellows in well-equipped, centrally-located laboratories, it is our goal that our trainees will be qualified to assume leadership positions related to transplantation and stem cell biology in industry, academia, and biotechnology. Our established track record demonstrates that we are well on our way to achieving this goal.
PI Name: Suzanne T. Ildstad
Title: Tolerance induction to islet transplants
Grant Number: R01DK069766
Fiscal Year Search: 2009
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7674545&icde=5639745
DESCRIPTION (provided by applicant):
The focus of this proposal is to develop a novel "conditioning" approach that will replace myelotoxic agents to establish chimerism in NOD mice. We will induce immune deviation to promote host-versus-graft hyporesponsiveness, thereby giving the hematopoietic stem cell (HSC) an opportunity to engraft and establish subsequent self-perpetuating deletional tolerance to islet allografts. Our recent studies in a mouse model suggest that the primary role for conditioning for HSC transplantation is to suppress host-versus-graft alloreactivity, rather than to prepare vacant niches in the recipient's bone marrow compartment. This observation suggests that one could replace myelotoxic agents with antigen-specific approaches to induce host-versus-graft hyporeactivity or anergy at the time of HSC transplantation. As the mechanisms underlying T cell activation are defined, highly specific approaches to suppress this alloreactivity have emerged. In AIM I. we will ESTABLISH CHIMERISM THROUGH IMMUNE DEVIATION OF THE RECIPIENT. We will immunomodulate the recipient: (a) targeting alloreactive cells in the host microenvironment; (b) inducing anergy and/or antigen-specific apoptosis of alloreactive host cells; and (c) through generation of regulatory T cells (Treg), and develop a novel nonmyeloablative conditioning regimen to induce antigen-specific hyporesponsiveness to the HSC and islet allografts. Cell-based therapies have great potential for inducing transplantation tolerance. Of greatest interest are the new subpopulations of bone marrow-derived dendritic cells (DC) that have recently been shown to be potently tolerogenic in vitro under certain circumstances. We are the first to demonstrate an in vivo engraftment-enhancing effect for precursor plasmacytoid DC (p-preDC). The exploitation of this discovery in vivo and its potential to reduce the need for myelotoxic conditioning has not yet been tested. Hematopoietic growth factors have also been used to drive the immune response to a tolerogenic T helper 2 (Th2) phenotype through production of p-preDC or other tolerance-promoting cells (graft facilitating cells {FC}) that in turn generate Treg. In AIM II, we will USE PRE-TRANSPLANT IMMUNOMODULATION OF THE DONOR WITH HEMATOPOIETIC GROWTH FACTORS TO GENERATE TOLEROGENIC CELLS IN THE HSC ALLOGRAFT. We will use these factors and the cells they generate to modulate the tolerogenicity of the donor marrow inoculum in vivo to tip the immune milieu in favor of graft acceptance, enhancing bone marrow chimerism without myelotoxic conditioning. We will examine the mechanism by which this occurs and identify which cell types in the graft are critical to tolerance induction. P-preDC exposed to apoptotic donor antigens are potently tolerizing in vitro through generation of Treg. The therapeutic application of this approach has not been tested in vivo. In AIM III, we will USE EX VIVO IMMUNOMODULATION OF THE MARROW to expand p-preDC and FC and induce a tolerogenic inoculum for HSC transplantation.
PI Name: Stuart Williams
Title: A Prevascularized Islet Immunoisolation Devcie
Grant Number: R01DK078175
Fiscal Year Search: 2010
http://www.projectreporter.nih.gov/project_info_description.cfm?aid=7786955&icde=5639745
DESCRIPTION (provided by applicant):
Beta-cell replacement therapy via islet transplantation remains a promising technology for the reversal of type 1 diabetes. A significant barrier to the clinical utilization of beta cell transplants has been the lack of a host-derived blood supply to maintain the viability and thus the function of transplanted cells. We have developed a new cell-based therapy for the generation of pre-vascularized tissue engineered constructs. We have also developed a new generation of biomaterials that support extensive neovascularization. The combined cell and material construct to be evaluated is termed a Prevascularized Immuno-Isolation Device or PVID. We propose to use these materials in the development of a new beta-cell immuno-isolation device to prolong beta cell viability and function. These constructs represent a pre-formed microcirculation that can be constructed from a patient's own fat-derived microvascular endothelial cells, avoiding the use of immuno- suppressive drugs. Specific aim 1 will evaluate the maturation of the microcirculation within a prevascularized construct following implantation in an animal model. Specific aim 2 will evaluate novel porous biomaterials and material surface modification to support the neovascularization of the porous material to assure perfusion of encapsulated islets. The biomaterial developed is a two component hybrid system that also provides immunoisolation for the encapsulated islets. Specific aim 3 will evaluate the viability and function of islets encapsulated in the prevascularized immunoisolation devices in an animal model of diabetes.