2007-08 Levinson Scholars

Joan now hails from Gig Harbor, WA, but has lived all over the United States. She has wanted to attend the University of Washington since she was 10 years old.

The summer of her freshman year, Joan was given an opportunity to perform research in the Dovichi Lab working with breast cancer cells, and has been researching ever since. She cannot say enough for the importance of research for any would-be-chemist. After all, knowledge is nothing without practical application.

Research has also given Joan the chance to present her work all over the country. She enjoys these trips immensely, even though she has a tendency to get incredibly nervous before speaking in front of others.

Joan’s chemistry career began with an amazing high school chemistry teacher. She believes it is important that young students get a good introduction to chemistry. Joan tutors at the University of Washington’s CLUE center and for the Youth Tutoring Program (YTP) in Jackson Park. After getting her degree in chemistry, she hopes to teach chemistry for a few years before attending graduate school to pursue her Ph.D.

Mentor: Norman Dovichi, Chemistry

Project Title: Protein fingerprinting of MCF-7 breast cancer cells

Abstract: Breast cancer is leading cause of cancer mortality for women in the United States. Prognostic indicators like progesterone receptor and Her2/neu proteins guide treatment and predict outcomes. Their expression levels directly affect patient survivability. Even with these indicators, there is a need for more detailed information to guide therapy. By generating protein fingerprints, which show the expression levels of multiple proteins at one time, our lab hopes to provide a valuable prognostic tool for cancer patients. MCF-7 is an established breast cancer cell line. By dosing MCF-7 with the chemotherapeutic agent mitomycin C (MMC), we hope to isolate survivor populations for protein studies. Cell viability will be determined by hemocytometry with Trypan blue staining and flow cytometry. Proteins in the survivor cell lines will be analyzed using 2D SDS PAGE/LC-MS and capillary electrophoresis with laser-induced fluorescence detection.

Following an army enlistment and two years at Bellevue Community College I transferred to the School of Oceanography at the University of Washington. The School of Oceanography has a very strong tradition of involving undergraduates in research, and I soon found a position in the lab of Prof. Julian Sachs. In spring 2007 I had the opportunity to help plan and conduct an expedition to a region of Canada known to contain many hypersaline lakes and to the Great Salt Lake of Utah. During this expedition I became very interested in the density and diversity of microbial life present within these harsh environments. After our return I began planning a project to enumerate and isolate extreme halophiles from some of our samples in order to identify a potential salinity indicator in the form of a lipid biomarker.

Mentor: Julian P. Sachs, Oceanography

Project Title: Enumeration and isolation of culturable halophiles from several hypersaline lakes and the development of a potential salinity indicator

Abstract: Halophiles recovered from hypersaline environments in western Canada and Utah’s Great Salt Lake provide an opportunity to advance our knowledge of how these organisms interact with their harsh environment. By treating these organisms to both aerobic and anaerobic, and light and dark conditions we can reduce competitive inhibition among culturable halophiles and enumerate the widest variety possible. Isolation of morphospecies cultured in this manner will enable us to determine the salt tolerances for individual species by inoculating pure colonies onto media of varying salinities. Analysis of lipid biomarkers produced by halophiles cultured at these varying salinities using gas chromatography mass spectrometry may reveal salinity indicators that can be applied to paleoclimate questions.

Lauren Hanson is interested in the development of the human brain and the causes and treatment of neurological disorders. Having entered the neurobiology program a year early, Lauren gained the background and skills to immerse herself in the research that she is most passionate about for the remaining three years of her program. Lauren’s mentor describes her as the top student in her cohort in the neurobiology program; she is someone who produces the highest quality work in every aspect of her education. In terms of the goals of the Levinson scholarship, he says: “I cannot imagine a better candidate than Lauren. She really is an ‘emerging scholar’ in the best sense of the award. Granting her this award will enable an unusually talented student to experience a real long-term research project in a way that few students have the opportunity to do.” Lauren’s award will provide her with scholarship support, research books and supplies and conference travel.

Mentor: Professor William Moody, Department of Biology, Chair of Neurobiology

Project Title: Physiological properties of pacemaker neurons driving spontaneous activity in the neonatal mouse brain

Abstract: Spontaneous electrical activity plays a central role in nervous system development. In mouse cortex, spontaneous synchronous activity (SSA) is vital for processes such as the formation of appropriate synaptic connections and development of normal neuronal properties. For SSA to carry out its developmental functions, it must occur during the appropriate critical stages of development. SSA must also cease to provide for the correctly timed emergence of the mature information-processing functions of neurons. Therefore, how the developmental timing and generation of SSA are controlled is a major question in neurobiology. It has been determined that SSA is run by a pacemaker region in mouse cortex. By utilizing calcium imaging, extracellular recording, and whole cell recording techniques on mouse brain slices, I plan to learn a great deal about the specific location, characteristics, development and control of the pacemaker region responsible for SSA. I will identify the population of neurons that compose the pacemaker region and begin by investigating its developmental emergence. From this stems my investigation of the unique properties of the pacemaker cells. Understanding the emergence of the pacemaker function and the ways in which it controls the onset, activity, and cessation of SSA is vital in understanding the mechanisms of brain development. Continued exploration of the pacemaker region also has potential to improve understanding of later development when it may contribute to the mechanisms of pathological forms of synchronous activity, such as seizures.

I first became interested in protein research during an honors biology seminar, studying the Ice Worm and the various adaptations that allow it to thrive in frozen environments. I was particularly intrigued by the protein modifications suited to low temperatures, structural variations that enhanced functionality in sub-zero climates. I was interested in further pursuing protein research, especially as it related to the medical field, and began working in the lab of Dr. Rachel Klevit. The Klevit Lab studies the breast cancer susceptibility protein BRCA1 using high resolution structural biology techniques such as Nuclear Magnetic Resonance (NMR). The research offers powerful insight into the structural and functional properties of BRCA1 and has profound implications in developing far more precise and effective cancer therapy.

My hope is to attend medical school and eventually pursue a career that incorporates both patient care and biomedical research. I am especially interested in pursuing cancer research, as the field is still just beginning to understand the complex biochemical interactions involved in such a wide array of pathologies. My current experiences in the Klevit lab studying BRCA1 are invaluable to that development. The knowledge and skills I have gained since beginning research are fascinating and have enhanced my understanding of biochemistry through intensified, hands-on application.

Mentor: Rachel E. Klevit, Biochemistry

Project Title: BRCA1-mediated ubiquitination

Abstract: The breast cancer susceptibility gene, BRCA1, encodes a crucial tumor suppressor protein involved in multiple DNA damage repair pathways. One of its most important functions is ubiquitin ligase activity, in which BRCA1 facilitates the transfer of the small protein ubiquitin to other protein substrates. The ubiquitin acts as a molecular tag, important for marking substrates in many cellular processes such as proteolysis, cell signaling, and DNA repair. The loss of ubiquitin ligase activity when BRCA1 contains cancer-associated mutations suggests an important role for this function in tumor suppression capacity. The various steps of ubiquitination also require adaptor proteins in addition to BRCA1 to successfully mediate ubiquitin transfer. The process by which the proteins interact, and why mechanistically the adaptors are necessary for transfer, is still unknown. The use of high resolution structural biology techniques, such as Nuclear Magnetic Resonance (NMR), offers powerful insight into the three dimensional characteristics of BRCA1-protein complexes. Structural elucidation will furthermore provide insight into the physical relationship and function of each interacting component, and expectantly lead to more precise treatment possibilities for breast cancer, the second most common cause of cancer related death in women.

My research experience started my freshmen year as I began working in the Applied Biomechanics Laboratory. We performed biomechanics testing on macaque monkey specimens in an attempt to understand the developmental changes in the cervical spine as a function of age. From there, I was invited to help engineer the force sensing glove I am currently involved with. It is an exciting opportunity to be creative and analytical while developing a device that will be practical and effective in physical medicine. The potential for this device to change the way clinicians obtain and share their results is exciting and motivating.

Mentor: David J. Nuckley, Mechanical Engineering

Project Title: Clinical Force-Sensing Glove

Abstract: For many clinicians, their effectiveness is dependent on the forces they apply to their patients. However, current care strategies lack quantitative feedback. My objective is to develop a force-sensing glove to provide real-time quantitative feedback to assist in clinical diagnosis and treatment. To minimally affect a clinician’s function, obtain maximal signal to noise in a medical environment, and maintain patient safety, I am currently developing a fiber optic sensor and methods for implementing it in a latex glove. When a fully functional instrumented glove is fabricated and calibration tests are performed, I plan on presenting the glove to various clinicians for a field test this spring.

Christine’s interest in medical research began with a histology project in high school and has deepened throughout her undergraduate years. She was her high school valedictorian, and won a prestigious Washington Scholars award for tuition at any college or university in the state as a high school senior. Her ultimate goal is to become a physician-scientist, linking clinical medicine and basic biology. She states: “I want to practice medicine with the analytical skills learned from research and to conduct research with human faces behind my motivation.” Her mentor ranks her among the very best students he has taught, speaks highly of Christine’s talent for research, and the critical timing of this award: “I believe that Christine has reached a defining moment in her undergraduate training as she embarks on an increasingly independent trajectory.” Christine’s award will provide support for research supplies and conference travel.

Mentor: Prof. Peter Rabinovitch, Department of Pathology

Project Title: Possible neuroprotective effects of mitochondrially-targeted catalase in the 3-NP model of acute oxidative stress

Abstract: There is considerable evidence that oxidative stress, resulting from exogenous and endogenous free radicals, has a causal role in aging and age-related disease, in particular, neurodegenerative diseases. The mitochondria have been shown to be the foremost site of endogenous free radical production. To further investigate this, we will study a transgenic mouse that expresses human catalase (an antioxidant enzyme) that is targeted to the mitochondria. This transgenic mouse demonstrates a 20% increase in life span and decreases in age-related pathology in the heart and possibly skeletal muscle. While there is research underway focusing on heart and skeletal muscle, this project will specifically relate to the brain, which has not yet been examined. One model that can test the importance of mCAT expression in the brain is the 3-nitropropionic (3-NP) model of acute oxidative stress. 3-NP is an irreversible inhibitor of succinate dehydrogenase, complex II in the electron transport chain of the mitochondria. 3-NP has been used by other investigators to recapitulate the clinical features of Huntington’s Disease, another age-related neurodegenerative disorder. These investigators have demonstrated that 3-NP produces selective striatal lesions that are attributed to neurotoxic effects of oxidative damage. In this study, we will investigate the possible neuroprotective effects of the mCAT compared to WT in the 3-NP model.

Lauren came to the UW for her undergraduate degree specifically because of the research opportunities offered for undergraduates. She has the unique experience of having been involved in research since the summer before her freshman year, working on two projects that explore cellular metabolism. She is very interested in a future that combines academic and biotechnology research and hopes to participate in academic-industry collaborations. Her mentor describes how Lauren has become an important member of her research group: “Not only has she significantly advanced her project, but she has also been instrumental in setting up and trouble shooting protocols for the lab…Her future work in uncovering the link between methanol metabolism and glutathione homeostasis is very important for our understanding of methylotrophy and the engineering of this organism for biotechnological and industrial use.” She adds: “She is one of the top students I have mentored and I have no doubt that she will have an outstanding scientific career and make a lasting contribution to science.” Lauren’s award will provide scholarship support, books, and conference travel.

Mentor: Professor Mary Lidstrom, Department of Chemical Engineering & Microbiology and Vice Provost for Research

Project Title: Metabolic Studies in Methylobacterium extorquens AM1

Abstract: Methylotrophs, or organisms that can grow on single-carbon compounds, have great potential for bio-industrial applications. Methylotrophs can be bioengineered to produce value-added products from methanol, a cheap and environmentally benign carbon source. Due to the interrelated nature of metabolism, successful engineering requires deep understanding of the organisms’ metabolism. We seek to further understanding of methylotrophy through study of the model methylotroph, Methylobacterium extorquens AM1. Building on the body of knowledge of methylotrophic pathways, my work seeks to elucidate links between methylotrophy and other metabolic networks. Previous results suggest strong links between methylotrophy and iron metabolism. My work will explore the differences in iron metabolism during methylotrophic and non-methylotrophic growth in AM1, including assaying iron requirements and characterizing iron-related proteins and mutants. This work will aid in the understanding of methylotrophic growth and general iron metabolism.

Teresa began her research experience she will was just a junior in high school. She was one of 14 students involved in a summer program called Lab Experience for the High School Student, which was created specifically to introduce high school students to the new and blossoming field of bioengineering. Since then, Teresa has been an active student researcher here at the University of Washington.

Her other activities include being a committee member for Relay for Life, and other cancer related organizations. She writes, “I regularly speak with the community about the importance of the work done by the American Cancer Society and how contributing to Relay for Life will affect the lives of millions of current and future cancer survivors. Before working with the American Cancer Society and Relay for Life, I had never understood just how devastating a disease cancer is.”

In the upcoming future, Teresa hopes to attend a graduate program where she can continue her research. She would like to seek a career in research where I can work towards making cellular and molecular treatments for cancer a clinical reality.

Mentor: Suzie Pun, Bioengineering

Project Title: Surface-modified Listeria-monocytogenes as a carrier for the intracytosolic delivery of therapeutics

Abstract: Macromolecule therapy, the introduction of therapeutic agents into diseased cells in the body, has the potential to revolutionize the treatment of life-altering diseases like cancer. One of the major needs in delivery to tumors is the development of carriers that can efficiently transport therapeutic agents into tumor cells. Listeria monocytogenes, a facultative intracellular pathogen, is a promising potential carrier because it is highly efficient at entering the cytosol after invading host cells. It is also capable of moving through and between cells, giving it the potential to make its way past the most accessible layer of the tumor, reaching deeper, central cancer cells for delivery. Attenuated strains of Listeria are of interest for intracellular therapeutics delivery. To deliver therapeutic genes to tumor cells, I propose to attach them to the surface of Listeria via a streptavidin-biotin linkage. I have developed a surface conjugation strategy that allows modular attachment of therapeutic cargo to Listeria. Listeria were surface-modified by reaction with NHS-biotin, enabling the attachment of biotinylated cargo through a streptavidin linker. Surface biotinylation of Listeria was confirmed through a plate-binding assay. Biotinylated Listeria bound to a plate displaying surface-adsorbed streptavidin, while unmodified Listeria failed to bind. The attachment of a biotinylated, fluorescently-labeled model protein cargo to the surface of biotinylated Listeria was examined, and the invasion of host cells by this Listeria-protein conjugate was investigated using fluorescence microscopy. I will further demonstrate delivery to the cell cytosol and begin investigation of cargo loading characteristics and cell cytotoxicity. This method will then be translated to a less pathogenic strain of Listeria and I will demostrate its ability to deliver antigenic peptides and stimulate T-cell activity. Listeria-drug conjugates in the future could be applied toward nucleic acid and protein delivery.