POSITION TITLE: Regents Professor and Head Medical Physiology, Director Div. of Lymphatic Biology

Transcription

1 OMB No and (Rev. 10/15 Approved Through 10/31/2018) NAME: David C. Zawieja, Ph.D. BIOGRAPHICAL SKETCH Provide the following information for the Senior/key personnel and other significant contributors. Follow this format for each person. DO NOT EXCEED FIVE PAGES. era COMMONS USER NAME (credential, e.g., agency login): ZAWIEJAD POSITION TITLE: Regents Professor and Head Medical Physiology, Director Div. of Lymphatic Biology EDUCATION/TRAINING (Begin with baccalaureate or other initial professional education, such as nursing, include postdoctoral training and residency training if applicable. Add/delete rows as necessary.) INSTITUTION AND LOCATION DEGREE (if applicable) Completion Date MM/YYYY FIELD OF STUDY University of Wisconsin, Green Bay, WI B.S. 05/1978 Biol/Chem/Pop Dynamics Medical College of Wisconsin Ph.D. 05/1986 Physiology Texas A&M University Post Doc 05/1988 Physiology A. Personal Statement: My lab group and I have been involved in studies on the microcirculation with a focus on the physiology of lymph transport for over 30 years. My expertise in the microcirculation includes studies on arteriolar myogenic constrictor function, regulation of vascular smooth muscle calcium, measures of venular permeability/function, and a lymphatic function. My lab has had a number of NIH projects (continuously funded since 1988) focusing on the role of lymphatic function in the microcirculation. Each of these projects has, as one of their objectives, the evaluation of the mechanisms (molecular, cellular, mechanical and tissue-level) regulating of lymphatic function. These projects focus on the ionic/calcium, smooth muscle contractile/regulatory protein, endothelialdependent shear-sensitive/no/cgmp pathways that are involved in regulating lymph transport, lymphatic muscle function and the interactions between immune cells, inflammatory agents and the lymphatics. To support this work we have established cultured cell lines of endothelial and muscle isolated from microlymphatics, acute and cultured isolated microlymphatic vessel preps, methodologies to evaluate lymphatic function at the single vessel, whole tissue and animal levels, measurement of lymph flow in microscopic lymphatics in situ, methodologies to target cell-specific gene manipulation in isolated lymphatic tissues, methods to study the role of NO in contractile function including methods to measure NO with microelectrodes and fluorescent indicators, methodologies to microscopically image in 3D/4D models of lymphatic network formation in the rat and mouse. We have used these skills and tools to study lymphatic function and dysfunction in a number of different pathologies including IBD, metabolic syndrome, infections, aging and other inflammatory states. B. Academic Positions Postdoctoral Res. Associate, Dept. of Medical Physiology, Texas A&M Univ Postdoctoral Fellow, Dept of Medical Physiology, Texas A&M Univ Asst Research Scientist, Dept of Medical Physiology, Texas A&M Univ Asst Professor, Dept of Medical Physiology, Texas A&M Univ Assoc Professor, Dept of Medical Physiology, TAMUSHSC 2003-present Director, Div. of Lymphatic Biology, Integrated Microscopic Imaging Lab, Dept. of Medical Physiology TAMUSHSC , 2013-present Professor, Dept of Medical Physiology, TAMHSC , Vice Chair, Dept of Medical Physiology, TAMHSC Interim Chair, Dept of Medical Physiology, TAMHSC present Head, Dept of Medical Physiology, TAMHSC 2013-present Texas A&M System Regents Professor

2 Honors NIH Natl Res Service Award, ; Travel Award, European Soc for Microcirculation, 1992; Intl Res Travel Award, Texas A&M Univ, 1994; NIH Res Career Dev Award, ; APS Frontiers in Physiology School Science Teacher Fellowship Award, Host, 1998; 2006 Representative for the Texas A&M Health Science Center at the Academy of Medicine, Engineering and Science of Texas Conference; Lymphatic Research Foundation Scientific Advisory board 2000 present, 2010 Lymphatic Research Leadership Award; Microcirculatory Soc Executive Council 2006-present, President ; Lymphatic Res and Biology Editorial board 2002 present, Microcirculation - Editorial Board 2003 present; 2010 Plenary Lecturer at the 61st Congresso Nazionale della Societa Italiana di Fisiologia, The D. Eugene Strandness Memorial Plenary Lecture at the 23rd Annual Meeting of the American Venous Forum, 2/2011. Plenary lecture at the International Research Center of Microvascular Medicine in the Institute of Microcirculation, Chinese Academy of Medical Sciences & Peking Union Medical College, 10/2011. Keynote speaker at the 10 th National Lymphedema Network International Conference, 9/2012Selected as one of the Texas A&M System Regent s Professors, 1/2013. Plenary lecture at the Joint Congress for the International Conference for Microcirculation, Chinese Association of Integrative Medicine, 9/2013. Awarded a Distinguished Visiting Professor Fellowship by the UK Royal Academy of Engineering to visit Imperial College, London, England. 5/2015 C. Contribution to Science I have identified some key peer-reviewed publications that highlight my most important contributions to science ( that helped shape the direction my lab has taken, influenced the work within the Lymphatic Biology Division and established important new paradigms in the understanding of lymphatic function as a critical player in the microcirculation. 1. The first of these papers Inhibition of the active lymph pump by flow in rat mesenteric lymphatics and thoracic duct. (PMID: ) demonstrates the important principle that fluid flow/shear inhibits the lymph pump and alters lymph transport and does so at shears much lower than previously thought to be active based on blood vessel studies, thus demonstrating important similarities and differences in lymph versus blood transport. Blood vessels have been known for many decades to be sensitive to the mechanical influences blood (i.e. pressure/stretch and flow/shear) has on them and lymphatics have been known to be sensitive to pressure and stretch. However, lymphatics had not been shown to be sensitive to the effects of shear on the lymphatic endothelium. In this study we demonstrated for the first time that lymphatic vessels were extremely sensitive to the effects of shear acting on the lymphatic endothelium. To do this we developed and implemented the isolated microlymphatic preparation in conjunction with my collaborators Drs. Michael Davis and Anatoliy Gashev. This has led to a series of papers by others and us that have subsequently focused on the biomechanical, cellular and molecular mechanisms the lymphatic shear sensitivity is dependent on. In these series of studies, we also developed and implemented the first methods to measure lymph flow in microlymphatics in situ and implement these parameters in the design of our isolated lymphatic preparations. These studies demonstrate how the lymphatic system effectively utilizes the lymphatic vessels themselves as pumps when it is needed to overcome inherent pressure gradients in the lymphatic network or diminishes that intrinsic pumping activity to allow extrinsic forces to drive lymph flow under the appropriate conditions. 2. The principal that the biomechanical sensitivity of the lymphatics is the key factor in the regulation of lymph transport set by the paper discussed above set the stage for further studies that lead to the second paper in my list of key contributions Regional variations of contractile activity in isolated rat lymphatics. (PMID: ). This paper demonstrates the important concept that lymphatic vessels from different regions of the body that are exposed to different fluid conditions and thus regulate lymph transport differently. This links that important principle of biomechanical sensitivity of the lymphatic to the structure and function of different lymphatic vessels in different tissues and animal models. To do this we conducted detailed characterizations of lymphatic structure and function using in situ studies in rat, guinea pig, mouse as well as isolated lymphatic studies in those same animal models as well as bovine, porcine monkey and dog studies. These studies have been commonly cited papers in the field, won the Microcirculatory Society 2004, Award for Excellence in Lymphatic Research and established paradigms that have been used in many other labs around the world. Many of these findings and the ones that came from the follow-up studies are the grounds for the basic principles of lymphatic transport that are the basis of the chapter I and my collaborators authored, Microlymphatic Physiology in the Handbook of Physiology: Microcirculation, that is used by and taught to students in ours and many other courses.

3 3. The 3 rd paper I use as an example of our most important scientific contributions is Nitric oxide formation by lymphatic bulb and valves is a major regulatory component of lymphatic pumping (PMID: ). This paper shows for the first time measurements of the production of nitric oxide by the lymphatic endothelium is differentially controlled by the lymph shear patterns in the lymphatic bulbs and valves. This is documented a gradient of NO production within the lymphatics structure that alter overall lymph NO levels, which alters lymphatic contractile function and may affect lymphatic muscle coverage and function as well as provide a potential chemo-attractive gradient for immune cells. This has led to a series of studies and papers investigating the role of NO molecular pathways in lymphatic function in the normal and pathological responses. 4. We have reinitiated studies started many years ago in my lab evaluating the regulation of intracellular calcium within vascular muscle and endothelial cells in response to mechanical and agonist activation. These studies started long ago in isolated arterioles ( Calcium measurement in isolated arterioles during myogenic and agonist stimulation, PMID: ) and dispersed vascular muscle cells ( Stretch-induced increases in intracellular calcium of isolated vascular smooth muscle cells, PMID: ). We have extended these to document the role of intracellular calcium levels in the regulation of lymphatic muscle cell contractile function ( Calcium sensitivity and cooperativity of permeabilized rat mesenteric lymphatics, PMID: and PKC activation increases Ca²+ sensitivity of permeabilized lymphatic muscle via myosin light chain 20 phosphorylation-dependent and independent mechanisms, PMID: ) and lymphatic endothelial cell shear response ( Measurement of shear stress-mediated intracellular calcium dynamics in human dermal lymphatic endothelial cells, PMID: ). This work demonstrated the important role that critical role that intracellular calcium plays in the function of vascular muscle and endothelial cells and that these roles are different in vessels from different parts of the microcirculation. 5. The next important contribution we have published, Molecular and functional analyses of the contractile apparatus in lymphatic muscle (PMID: ) demonstrated for the first time that the unique functional characteristics that lymphatic exhibit as both a vascular system and fluid pump are reflected in the molecular characterization of lymphatic muscle as a hybrid of smooth and striated muscle elements. This involved incorporating genetic, cellular and molecular techniques that required the expertise of one of my now longstanding colleagues and Multi-PI of this application, Dr. Mariappan Muthuchamy. Through our collaborations we developed and implemented many of the techniques commonly employed in cellular experiments (western blots of isolated vessels proteins, RT and qpcr of mrna from isolated microlymphatics to the significantly more challenging isolated lymphatic studies. The novel findings detailed that the bimodal functional properties of the lymphatic vessels being both a conduit and a pump are reflected in the molecular players involved in the contractile apparatus of the lymphatic muscle. This detailed for the first time that the tonic contractile functional character of the lymphatics is matched with classic fast/phasic smooth muscle contractile and regulatory proteins, while the same cells also have the striated contractile and regulatory proteins commonly found in cardiac and skeletal muscle. This linked the well-known functional and biomechanical properties that have been described for decades in lymphatics with a unique combination of smooth and striated contractile proteins. This is also linked to some of the upstream mechanisms we have also studied regulating the phasic lymphatic pumping the electrophysiological activity driving the pace making and action potential necessary for lymphatic phasic contractions and the subsequent increases in intracellular calcium needed for both the phasic lymphatic pumping and chronic lymphatic tone that we and others are now currently studying. 6. The lymphatic network is a complex system of vascular and immunological components that must act together in concert with other body elements to accomplish the main functions of the lymphatic system. Understanding these complex interactions is served well by developing appropriate computational models of them. My lab has developed and maintained strong collaborations with members of the engineering school in an effort to conduct modeling of lymphatic function. This has resulted in a series of papers, that model the elements of the lymphatics system from the molecular level to the scale of complex vascular network. An example of this work is depicted in our recent paper, Network Scale Modeling of Lymph Transport and Its Effective Pumping Parameters (PMID: ). 7. The lymphatic transport system is critically important to not only the transport of fluid, but also the transport of large molecules and cells/particulate matter. It is anatomically poised and functionally empowered to serve as the immune systems information pathway. Indeed, we have described the lymphatic vascular network as an immunocentric vascular network poised to transport immune-relevant signals from the parenchymal tissues of the body to the secondary lymphoid organs (lymph nodes) to regulate the body s immune response to self and foreign antigens. To this end we have recently published a series of papers

4 that document and highlight these interactions between immune and fluid responses: ( An immunological fingerprint differentiates muscular lymphatics from arteries and veins PMID: ; Collecting lymphatic vessel permeability facilitates adipose tissue inflammation and distribution of antigen to lymph node-homing adipose tissue dendritic cells. PMID: ). 8. Inflammatory responses are the classic signals of tissue injury/damage that start the transition from normal immunological self-tolerance towards foreign antigen responses. Historically lymphatics re know to play key roles in the body s response to inflammation. We have published numerous studies that have evaluated the effects of inflammatory signals on various elements of lymphatic function: ( The effects of inflammatory cytokines on lymphatic endothelial barrier function. PMID; Lipopolysaccharide modulates neutrophil recruitment and macrophage polarization on lymphatic vessels and impairs lymphatic function in rat mesentery. PMID; Effects of f-met-leu-phe-induced inflammation on intestinal lymph flow and lymphatic pump behavior. PMID: ; Effects of substance P on mesenteric lymphatic contractility in the rat. PMID: ) 9. The impact of inflammation on lymphatic function and vice versa has lead our group to the study of lymphatic function under various pathological models of inflammation: ( Colonic Insult Impairs Lymph Flow, Increases Cellular Content of the Lymph, Alters Local Lymphatic Microenvironment, and Leads to Sustained Inflammation in the Rat Ileum PMID; Macrophage alterations within the mesenteric lymphatic tissue are associated with impairment of lymphatic pump in metabolic syndrome PMID: ) 10. The last important contribution to science I wish to highlight is our series of studies on the impact of the space environment of lymphatic function. Lymphatic vessels are particularly sensitive to small changes in pressure and their function are heavily affected by the gravitational influences in hydrostatic pressure. Astronauts in long-term space flight experience symptoms of lymphatic dysfunction, upper body edema, regional inflammation, immune dysfunction etc. We have utilized both land-based microgravity models and animals flown in space to study the changes that occur in lymphatic function under the conditions experience in long space flight. ( Inhibition of active lymph pump by simulated microgravity in rats. PMID ). Complete List of Peer-reviewed Publications in MyBibliography: D. Research Support ONGOING NASA Research Announcement (NRA) NNH14ZTT001N Spaceflight Research Opportunities in Space Biology, 14-14SF_Step2-0079, (Zawieja, D PI, Cromer, W. CoI.) 9/2014-9/2016 Effects of microgravity on lymphatic structure and function (#14-14SF_Step2-0079) The central hypothesis is that long-term exposure to microgravity during space flight induces the development of a low level chronic inflammation of the gut, alters lymph flow, lymphatic proliferation and lymphangiectasia and the transport of inflammatory cells in the lymph. Our group will specifically study the impact of space flight on function of the mesenteric lymphatics and gut inflammation. NIH-NBIB/NHLBI U01HL123420, (Zawieja, D. MPI, Moore, J. MPI) 09/ /2019 Transport Phenomena in the Lymphatic System The goal of this grant is to develop a multiscale model of lymph transport using data derived from mouse and rat tissues. NASA, NNH12ZTT001N Research Opportunities in Space Biology. (Delp-PI; Zawieja- CoPI) Effects of Microgravity on Cerebral Arterial, Venous, and Lymphatic Function: Implications for Elevated Intracranial Pressure The central hypothesis is that long-term exposure to microgravity during space flight alters the balance of forces regulating intracranial pressure. Our group will specifically study the impact of space flight on function of the cervical lymphatic that help regulate CSF pressure. NIH R15, HL (Montamas Suntravat-PI; Zawieja-Consultant) 11% score pending Council review The Acute Effects of Snake Venom CRiSP Toxins on Blood and Lymphatic Endothelial Cell Permeability: New Insights into the Pathology of Snakebite The central theme of this project is to evaluate the effects of components of rattlesnake venom (CRiSP) on the gross edema associated with rattlesnake bites.

5 Grants Completed In The Past Three Years NIH R01, DK09922, (Zawieja-MPI); Muthuchamy-MPI) Role of mesenteric lymphatics and dietary endotoxin in metabolic syndrome The central theme of this project is to study the changes in lymphatic function during the inflammation induced during the development of metabolic syndrome in rats by high fructose feeding. NIH-NIDDK Admin. Supp. - Women s Health Initiative, (Zawieja-MPI, Muthuchamy MPI) 9/2014-8/2015 Role of mesenteric lymphatics and dietary endotoxin in metabolic syndrome The central theme of this project is to study sex-based differences in the changes in lymphatic function during the inflammation induced during the development of metabolic syndrome in rats by high fructose feeding. NIH-NCI, R01 (Ran PI; Zawieja CoI) NF-kB mediated induction of VEGFR-3 and new lymphatic vessels in breast cancer This project evaluates how the inflammatory agent NF-kB induces expression of VEGFR-3 and stimulates lymphangiogenic growth. NIH, R01 HL (Zawieja MPI; von der Weid Univ of Calgary MPI) Ionic Mechanisms of Stretch-Activation In Muscular Lymphatics The central theme of this project is to study the ionic mechanisms responsible for the regulation of lymphatic contractility via stretch-activation. NIH R01 AG (Gashev PI; Zawieja - CoI) 08/15/08-07/31/14 National Institute of Aging Mechanisms of the age-related alterations in lymphatic pumping The central hypothesis of this proposal is that aging impairs both pressure and flow-dependent regulatory mechanisms in lymphatic vessels. This will result in a regression of the functional adaptive reserves in lymphatic contractility due to enos/inos imbalances, along with alterations in crossbridge activation mechanisms in lymphatic vessels.