The following departmental faculty were awarded competitive and non-competitive research grants during calendar year 2012.
Dr. Jeff Taekman was awarded unrestricted medical education grants totaling $750,000 from Pfizer and Abbott to support a project focused on improving clinical handovers. A broad range of departmental, institutional, and extramural collaborators will work together to improve healthcare professional competence and performance. The simulationbased initiative features a multi-modal educational strategy (including the use of virtual environments as well as in-situ observation / coaching). Key collaborators include Ryan Fink, Noa Segall, Jonathan Mark, and David Turner (Pediatric Critical Care).
Madan Kwatra, PhD, Associate Professor of Anesthesiology in the Division of Basic Sciences received a 2-year $403,250 NIH Exploratory / Developmental Research Grant Award (R21) from the National Institute of Neurological Disorders and Stroke (NINDS) entitled “Truncated NK1R in GBM: Pharmacology and Relationship with Patient Survival”. Glioblastoma multiforme (GBM) is a deadly brain tumor and patients diagnosed with GBM die within 2 years. Current treatments for glioblastomas include surgery, radiation therapy, and chemotherapy, but these are ineffective and we need to develop new ways to block GBM growth. Our laboratory has shown that NK1R is a good target to inhibit the growth of GBM and the proposed studies will determine the pharmacology of the truncated form of NK1R and whether its expression in GBM tumor predicts patient survival.
Luke James, M.D., Assistant Professor in the Division of Otolaryngology, Head, Neck/Neuroanesthesiology, along with Christopher Kontos, MD and Nicholas Katsanis, PhD (Department of Medicine) were awarded a $50,000 2012 Department of Medicine Integrated Research Award entitled “Regulation of Inflammation and Vascular Permeability in Intracerebral Hemorrhage”. Intracerebral hemorrhage (ICH) causes 15% of strokes, but only 20% of patients with ICH are functionally independent six months after the event. Currently, supportive care is the primary treatment for ICH, but new evidence finds that inflammation and vascular permeability contribute to neurological dysfunction after ICH. Christopher Kontos, MD, associate professor of medicine (Cardiology), Nicholas Katsanis, PhD, professor of cell biology, and Michael “Luke” James, MD, assistant professor of anesthesiology and medicine (Neurology), will examine the endothelial receptor tyrosine kinase Tie1 in inflammation and permeability after ICH. After considering more than 50 proposals submitted to the new Department of Medicine Integrated Research RFA initiative, the Department of Medicine Chair’s Office has awarded funding to eight projects.
Joseph P. Mathew, MD, MHSc, Professor of Anesthesiology and Chief of the Division of Cardiothoracic Anesthesia and Critical Care Medicine (along with the Co-PI Dr. Jeffrey Browndyke in the Department of Psychiatry and Behavioral Sciences) received a 2-year $431,750 NIH Exploratory/Developmental Research Grant Award (R21) from the National Heart, Lung, & Blood Institute (NHLBI) entitled “Functional Connectivity Changes in Attention Networks After Cardiac Surgery”.
Noa Segall, PhD, Assistant Professor of Anesthesiology and human factors engineer in the Human Simulation and Patient Safety Center, received a 1-year $99,989 AHRQ Small Research Grant Program (R03) entitled “Workload Effects on Response Time to Life-Threatening Arrhythmias”. To increase the potential for timely detection and treatment of in-hospital cardiac events, more and more at-risk patients are now monitored remotely by cardiac telemetry technicians. However, decisions regarding the appropriate number of patients that a single technician may safely and effectively monitor are largely based on technological capabilities and not on our understanding of human information processing limitations. We propose to use high-fidelity simulation of cardiac telemetry to determine the impact of increasing the number of patients monitored on response time to cardio-respiratory events. The knowledge to be gained will inform efforts to study this problem in real world cardiac telemetry and, ultimately, help to develop evidence-based standards for remote monitoring.
Congratulations are in order for Wulf Paschen, PhD, Professor of Anesthesiology in the Division of Basic Sciences for his application entitled “Proteomics Analysis to identify SUMO-conjugated proteins in tissue extracts” that was submitted to the Duke University School of Medicine Core Facility Services Voucher Program for a services voucher totaling $10,000. A large number of applications were submitted and this voucher for services enables Dr. Paschen to identify SUMO-conjugated proteins in tissue extracts. Dr. Wei Yang in Dr. Paschen’ lab generated a new highly innovative mouse model conditionally expressing tagged SUMO paralogues with which it will be possible, for the first time, to advance SUMO proteomics from cell cultures to organs.
Congratulations to Quintin Quinones, MD, PhD, CA-2 Resident and ACES Scholar, along with his mentor Dr. Mihai Podgoreanu (Division of Cardiothoracic Anesthesiology) on their $75,000 Research Fellowship Grant from the Foundation for Anesthesia Education and Research (FAER) for their proposal titled “Organ Protective Mechanisms Invoked in Mammalian Hibernation”. Ischemia-reperfusion injury (I/R) is an unavoidable consequence of cardiac surgery and organ transplantation, and remains a major contributor to peri-operative mortality. The aim of this proposal is to investigate the transcriptional, protein expression, and metabolic differences in the hearts of arctic ground squirrels compared with non-hibernating mammals and to identify new targets to reduce ischemia-reperfusion (I/R) injury in human disease states. This award speaks to the success of the Duke ACES program in mentoring and supporting talented residents who aspire to careers in academic anesthesiology and seek to supplement their clinical training with opportunities for research and innovation.
Quintin Quinones, MD, PhD and Mihai Podgoreanu, MD were awarded $10,000 from the Duke University School of Medicine Core Facility Services Voucher Program for an application titled “Cardioprotective mechanisms invoked in mammalian hibernation: the role of innate immunity”. The grant will enable preliminary studies on the role of danger associated molecular pattern molecules released by tissues injured during ischemia in activating antigen presenting cells that circulate upon reperfusion, and mediating a second wave of inflammatory injury. Specific differences in innate immune responses that underpin the cardioprotective hibernator phenotype will be tested using flow cytometry and proteomic approaches.
Wei Yang, PhD, Assistant Professor in the Division of Basic Sciences was awarded a fouryear $308,000 NCRP Winter 2012 Scientist Development Grant from the American Heart Association (AHA) entitled “Trans-genic Mice to Uncover the Role of SUMOlyation in Brain Ischemia/Stroke”. Research results from a study sup-ported by a DREAM Innovative Grant funded by the Department of Anesthesiology provided the preliminary data for the AHA grant proposal. This project is Dr. Yang’s first extramural funding. Small ubiquitin-like modifier (SUMO1-3) is a small group of proteins that are conjugated to lysine residues of target proteins and thereby modify their activity, stability and subcellular location. The SUMO conjugation pathway modulates various pro-cesses playing important roles in key cellular functions under normal and pathological conditions. These func-tions include gene expression and genome integrity, proteasomal degradation of proteins, quality control of newly synthesized proteins, and DNA damage repair pathway. Dr. Yang has demonstrated that both transient focal and global cerebral ischemia massively activates SUMO conjugation. However, the role of the ischemia-induced rise in levels of SUMO-conjugated proteins in the fate and function of post-ischemic neurons has never been established owing to the lack of experimental tools to manipulate this pathway in the intact brain. Dr. Yang together with the research team of the Paschen laboratory has generated unique novel SUMO transgenic mouse models with which it will be possible to uncover the mechanisms that link the post-ischemic activation of SUMO conjugation to the fate and function of neurons exposed to a transient interruption of blood supply. Dr. Yang is expecting that the planed project will uncover the role of SUMO conjugation for the life/death decision of post-ischemic neurons and help to identify the main players in this process, i.e. proteins that are SUMOconjugated after ischemia. Ultimately, such knowledge has the potential to translate into novel avenues of therapeutic interventions in pathological states associated with ischemia and various disorders associated with the SUMO conjugation pathway, including degenerative diseases, cancer and diabetes.
Thomas Buchheit, MD, Associate Professor in the Department of Anesthesiology, Divisions of VAMC and Pain Management, along with Drs. Andrew Shaw and Tom Van de Ven, have received notice of award of a $2.9 million Department of Defense Grant over 4 years to study the role of epigenetics in the transition from acute inflammatory pain to chronic neuropathic pain. The grant, PT110575, titled “Regional Anesthesia and Valproate Sodium for the Prevention of Chronic Post-Amputation Pain” uses a clinical trial and nested metabolomics, epigenomics and gene expression analysis to dissect mechanisms that may prevent the development of chronic pain.
The Duke/VA research group will partner with their collaborators at Walter Reed to run a clinical trial at both the Durham VA and Walter Reed in active duty and veteran amputee patients. “This is full justification of our decision to focus on amputation as a model system for understanding persistent postoperative pain states, and we look forward to expanding our current amputation pain clinic to handle the increased workload this grant will bring” said Dr. Buchheit.
Dr. Shaw noted that this is the second large federal grant the VA group has been awarded in the past 18 months, and said: “We are delighted to continue our existing highly successful relationship with DOD, and fully support their goal-oriented approach to funding important translational medical research. Understanding how we respond to environmental trauma and stress is a key component of our research program, and this award fully cements our group at the cutting edge of inflammatory epigenomics.”
The award brings the group’s total federal active support to $4.4 million dollars, and will be conducted in their laboratory in GSRB1, the Duke Epigenetics and Epigenomics Program (of which all three are faculty members), the Durham VAMC and at Walter Reed National Military Medical Center, in collaboration with Col. Chester “Trip” Buckenmaier.
Wulf Paschen, PhD, of the Division of Basic Sciences, received a 2-year $156,000 research grant from the National Institute of Neurological Disorders and Stroke entitled “Restoration of Endoplasmic Reticulum Function in Experimental Stroke”. Transient cerebral ischemia/stroke impairs endoplasmic reticulum (ER) function, which in turn triggers translation arrest that is irreversible in vulnerable neurons. Cells cannot survive when ER function is irreversibly damaged by a severe form of stress. For functional recovery of neurons following a transient interruption of blood supply, restoration of ER function is therefore as crucial as reperfusion of the ischemic tissue. To cope with conditions associated with ER dysfunction, cells have developed a highly conserved stress response known as the unfolded protein response (UPR). UPR is characterized by activation of three ER membrane-resident proteins, PERK, IRE1 and ATF6. Upon ER stress, ATF6 translocates to the Golgi membrane where it is cleaved into a soluble form that translocates to the nucleus and activates expression of genes coding for proteins involved in the folding and processing of newly synthesized proteins within the ER lumen. The aim of the proposed studies is to focus on ER function as a therapeutic target by generating transgenic mice for transient cerebral ischemia/stroke experiments, using a genetic construct by which expression of ATF6-dependent genes can be activated in a tissue- and cell-specific fashion after crossbreeding with the respective Cre transgenic animals and induction by exposure to tamoxifen. Results of the proposed studies will provide key information on the new strategy of forced ATF6 expression to make neurons more resistant toward a transient ischemic stress. Impairment of ER function is believed to contribute to various pathological states of high clinical relevance, including ischemia, brain and spinal cord trauma, diabetes, and degenerative diseases. Creating ATF6 transgenic animals will therefore provide a key platform for future studies to establish the role of ER dysfunction in various clinically relevant pathologies.
Ru-Rong Ji, PhD, of the Sensory Plasticity Laboratory in Division of Basic Sciences, received a new 5-year $1,954,875 NIH Research Grant (R01) from the National Institute of Dental and Craniofacial Research (NIDCR) for his research project entitled “Hemichannels, astrocytic release, and neuropathic pain”. Pain conditions are a major health problem in the US and lead to medical morbidity and a reduced quality of life for millions of Americans. Chronic neuropathic pain conditions are especially difficult to treat. A largely unaddressed challenge is how the transition from acute pain to chronic neuropathic pain occurs and how to prevent and reverse this transition in patients. Spinal cord synaptic plasticity and long-term potentiation (LTP) have been strongly implicated in chronic neuropathic pain development. Accumulating evidence also points to an important role of glial cells in the pathogenesis of neuropathic pain. Astrocytes are the most abundant cell type in the CNS and maintain the homeostasis of the CNS. It is well established that astrocytic hemichannels such as connexin- 43 (Cx43) constitute an important pathway for gliotransmitter release. Although Cx43 was typically thought to regulate gap junction communication between astrocytes, this function could be switched to paracrine signaling via ATP and glutamate release under injury conditions. Our central hypothesis is hemichannels-mediated gliotransmitter release after nerve injury contributes to transition from acute pain to chronic neuropathic pain by modulating spinal cord synaptic plasticity and LTP.
Wulf Paschen, PhD, Professor of Anesthesiology in the Division of Basic Sciences, received a 5-year $1,717,190 NIH Research Grant (R01) from the National Institute of Neurological Disor-ders and Stroke (NINDS) for his research project entitled “Role of SUMO Conjugation in Ische-mia: Significance, Mechanisms, and Pathways”.
The project was funded in the first round of submission. Working with Dr. Paschen on the project are Co-Investigators Drs. Wei Yang and Huaxin Sheng from the Department of Anesthesiology, with assistance from Dr. Will Thompson in the Institute for Genome Sciences and Policy (IGSP) who will perform quantitative LC-MS/MS-based SUMO and ubiquitin prote-omics analyses. Small ubiquitin-like modifier (SUMO) conjugation modulates all major cellular pathways, including those associated with gene expression and genome stability, protein quality control, proteasomal degradation of proteins and DNA damage repair. Transient cerebral is-chemia massively activates SUMO conjugation, resulting in a dramatic rise in levels of SU-MO2/3-conjugated proteins. Cell culture studies suggest that the post-ischemic activation of SUMO2/3 conjugation is a protective stress response. However, the role of SUMO conjugation in the fate of post-ischemic neurons in the intact brain and the mechanisms and pathways that link SUMO conjugation to restoration of function impaired by transient ischemia are not known. Without this knowledge it is highly unlikely that the SUMO conjugation pathway can be manipu-lated for therapeutic purposes. Our long-term goal is to understand how to manipulate the SUMO conjugation pathway for preventive and therapeutic purposes. The objective of this particular application is to elucidate the role of individual SUMO paralogues in the recovery of neurons from ischemic stress and to identify the mechanisms and pathways involved.
During the past 25 years, our laboratory has been interested in understanding the mechanisms linking stress responses to the fate of cells exposed to stress conditions with focus on cerebral ischemia/stroke, a pathological state of high clinical significance. Since transient cerebral is-chemia/stroke triggers a long-lasting suppression of protein synthesis, we have focused on post-translational protein modifications. SUMO conjugation is believed to play key roles in the physiology and pathology of cells because it modulates all major cellular pathways. The com-plexity of processes modulated by SUMO conjugation is a challenge because the tools needed to investigate the role of SUMO conjugation in pathological states have not been available. In-deed, the knowledge that SUMO conjugation is mechanistically linked to key pathways that are essential for cellular integrity, including gene expression and genome stability, protein quality control, proteasomal degradation of proteins, and DNA damage repair is based almost entirely on cell culture studies, many of which were performed on yeast cells. When the tools to investi-gate the role of SUMO conjugation in pathological states are at hand, we will have a unique op-portunity to open a new dimension in SUMO research and advance from cell culture models to intact organs.
The project is highly innovative, because it uses, for the first time, SUMO3-/-, conditional SU-MO2-/- and SUMO2-/-SUMO3-/- mice, and SUMO transgenic animals that conditionally express tagged SUMO paralogues. These mouse models were developed and are in the process of be-ing developed in our laboratory and will allow us to elucidate the significance of SUMO conjuga-tion in post-ischemic brains. This will also be the first study to identify proteins that are SUMO- or ubiquitin-conjugated after ischemia, the first in vivo study to investigate the crosstalk between the SUMO and ubiquitin conjugation pathways, and the first comprehensive in vivo analysis of the protein-protein interaction networks, biological processes, and canonical pathways that link mechanistically SUMO conjugation to the fate and function of cells exposed to stress conditions.
Brain ischemia, brain tumor, and brain ageing/degenerative diseases have a common denomi-nator that defines the fate of cells – stress response pathways. Various stress response path-ways that make cells more resistant to damage induced by ischemia are also activated in cells in a solid tumor and help these cells to survive unfavorable environmental conditions, while ageing is believed to be associated with decreased capacity of cells to respond to stress. Our over-arching goal for future studies is to uncover the mechanisms that define the role of the SUMO conjugation pathway in these three closely related pathological states that have become re-search priorities as our population ages. In addition to ischemia, the SUMO conjugation is asso-ciated with various human diseases including, cancer, diabetes, degenerative diseases, arthritis, and heart failure. Our findings from the planned studies will therefore contribute to a better understanding of the mechanisms that link SUMO conjugation to these disorders.