Thomas Yorio - US grants

DESCRIPTION (provided by applicant): This is a competitive renewal of the Short-Term Research Training for Minority Students that we entitle "Summer Minority Advanced Research Training (SMART)". This proposal continues a highly successful effort to encourage and recruit underrepresented minority undergraduate students to pursue careers in the biomedical sciences by providing a ten week summer traineeship in a research environment that focuses on chronic diseases that affect minority populations (e.g. cardiovascular disease, diabetes, hypertension and glaucoma). This program involves a multidisciplinary approaching incorporating faculty from among the biomedical science division of the health science center. Trainees attend workshops and lectures on: Laboratory Safety and Practice, Animal handling and Animal Rights, Scientific Communications and Health disparity issues. Students will take a class on "Introduction to cardiovascular science" and receive two hours of credit toward their degrees at their home institutions. Each student selects a research mentor from the participating faculty after reviewing their research programs and listening to faculty presentations. Students and faculty design and plan a research project that can be accomplished within the ten-week period. All students must complete an abstract suitable for submission and presentation at the National Minority Research Symposium that is held each fall. Students gain experience in 1) generating an hypothesis; 2) testing their hypothesis through research experimentation; 3) using new research methodology; 4) evaluating and analyzing data using computer techniques, and 5) presenting their findings in a written abstract and oral presentation at the end of the training period. Students also receive lectures and review case studies on the responsible conduct of research early on in their training program. The overall effect of these activities is to create a sense of becoming a member of the scientific research community. This sense is furthered by student participation in their mentor's research group and departmental weekly seminar meetings. Students and faculty mentors each evaluate the program and each other at the end of the program. An external evaluator is used to help assess the program process as well as program outcomes. Students will be tracked throughout their participation in the program and after by formal mailed questionnaires, email follow-ups and the use of a list serve. Students will be recruited to return to the McNair summer program, if still eligible, and for entry into graduate school. Success will be measured ultimately by the number of students who enter biomedical science graduate programs and research as careers.

This proposal requests funding for a state-of-the-art digital transmission electron microscope, the Zeiss EM 910. This instrument will serve a growing community of researchers at the University of North Texas Health Science Center at Fort Worth as well as a number of other researchers from local institutions. Currently the electron microscopy lab of the Microscopy Core Facility serves 12 researchers. This includes 5 major projects with the remainder being comprised of minor projects, graduate students and non-Health Science Center researchers. The Microscopy Core of the Health Science Center is housed within the Department of Anatomy and Cell Biology and is currently attempting to upgrade its facilities. Several pieces of state-of-the-art support equipment have already been acquired. The Zeiss EM 910 will replace an aging transmission electron microscope which cannot be upgraded to the levels of sophistication and ability currently needed. The Zeiss EM 910 is a state-of- the-art instrument which will enable us to meet our current and future needs, as well as add an integral piece of equipment to an ongoing effort to improve the research infrastructure at this new health science center.Our current research needs, which the Zeiss EM 910 meets and our present microscope does not meet, include: 1. Improved image contrast for work with low contrast specimens (i.e. immunocytochemistry and in situ hybridization). 2. The ability to record, store, manipulate and print digital images from the transmission electron microscope. 3. The ability to perform morphometry, stereology and particle analysis using digital images. 4. Storage and automated recall of specimen grid coordinates. 5. Low magnification micrography and automated montaging. 6. Enhanced data recording to both negatives and archival computer files. 7. Utilization of the Health Science Center's computer network for exchanging data and images between researchers here an d around the country. 8. Electronic image rotation to allow better orientation of images prior to recording. 9. Future upgradability to cryoelectron microscopy, scanning transmission electron microscopy, energy dispersive x-ray analysis and the in-line Omega electron energy filter. This instrument will also compliment our goal of improved training of institutional and community graduate students and.technicians. It will also enhance the current medical profession program and the planned allied and public health professions programs of this new health science center.

The Student, Teacher Applied Research Training (START) project incorporates a one-on-one research apprenticeship for high school students and K-12 teachers, while providing an opportunity for teachers to develop a science laboratory to incorporate into the science curriculum in the Fort Worth Independent School District (FWISD). This grant will provide the basis for an interactive partnership of professional biomedical scientists and their graduate students at the UNT Health Science Center (HSC) with the teachers and students of the FWISD. The program involves an interdisciplinary approach utilizing faculty from the Graduate School of Biomedical Sciences. The program is designed to accommodate trainees with varied backgrounds by providing a 2 hr twice weekly course organized to present the basics of some new areas of biomedical sciences. Students and teachers will work closely with faculty mentors on ongoing research projects in their laboratories and will be required to present their findings at the end of the summer in an oral presentation and written report. HSC faculty will assist teachers in the development and implementation of a laboratory experiment based on their summer research experience that is suitable for inclusion into the FWISD science curriculum. A quarterly in-service science teacher forum will also be conducted by the program directors in conjunction with the START teachers for the purpose of continued education and to demonstrate the laboratory experiments developed by the START teachers for dissemination district wide. Students, teachers and mentors will evaluate the program at the end of the summer training period for use in subsequent planning. This program will provide a stimulus to students to seek careers in science by introducing them to the scientific method and the excitement of doing research. Teachers should gain a heightened awareness of the new areas of biomedical sciences and feel confident in including this information in their classroom curriculums.

DESCRIPTION (From the Applicant's Abstract): Glaucoma is a leading cause of blindness in the U.S. and worldwide that occurs as a result of a loss in retinal ganglion cells and a progressive optic neuropathy. Identification of the causative factors leading to this disease is of great interest. Recently, it has been shown that patients with primary open angle glaucoma (POAG) have an elevation in their aqueous humor endothelin (ET) concentrations. Similarly, in a animal model of glaucoma (Beagles) a four-fold increase in endothelin is seen in the aqueous humor. Our laboratory has been investigating the role of ocular endothelins for several years and recently have shown that it may be playing a role in the pathogenesis of glaucoma based on preliminary observations of its effects on axonal transport and ET's ability to activate the NOS-2/NO pathway. The overall goal of this application is to demonstrate that endothelin is an important ocular derived peptide that is not only involved in IOP and vascular homeostasis, but is a key component in the pathogenesis of glaucoma. The hypothesis to test is that ET is preferentially synthesised and released in the eye and at increasing concentrations mediates a series of events that exert detrimental effects on optic nerve function through decreases in axonal transport and increases in neurotoxic compounds, such as NO. The net effect is to promote retinal ganglion cell death. The following aims are planned to address this hypothesis: 1) Can elevated pressure or neuronal regulation increase endothelin synthesis and release in retinal pigment epithelium (RPE), optic nerve astrocytes (ONA) and lamina cribosa (LC) cells? Is there a functional cross-talk between ET and nitric oxide (NO)? 2) What signal transduction mechanisms and transcriptional regulation sites are associated with ET synthesis following elevation of pressure, neuronal regulation or activation of the NO pathway in RPE, ONA and LC cells? 3) Does ET play a role in the optic nerve damage observed in the in vivo rat elevated pressure model? Are there changes in ET receptor expression in various ocular tissues following elevation of IOP and is there a correlation between ET aqueous concentrations to elevations in IOP? and 4) Determine the ET receptor subtype involved and the role of NO in the optic nerve damage and effects on axonal transport induced by elevated pressure or ET administration. These specific aims are designed to determine if ET contributes to the optic neuropathy associated with elevations in IOP. Ultimately these studies could provide insight into the etiology of glaucoma and propose new alternative treatments for patients with glaucoma.

DESCRIPTION (provided by applicant): The University of North Texas Health Science Center is committed to the training of a diverse biomedical sciences workforce through the Graduate School of Biomedical Sciences masters and doctoral programs. Through a Coordinated Plan that links K-12 (Adopt-A-School) to undergraduate (SMART, McNair) and graduate (NIH MKITS and NSF K-12 Graduate/Fellow teacher programs) training in the biomedical sciences, the graduate school has achieved >20% underrepresented enrollment over the past several years. However, such accomplishments are still short of the need to develop parity that is reflective of the underrepresented population, particularly for Hispanics in Texas. The current IMSD Minority Opportunities in Research and Education (MORE) proposal addresses this need through the following goals: Goal 1 - Increase the number of underrepresented minority students entering graduate programs at UNTHSC to approach that of the Texas demographic profile of these groups and Goal 2 - Retain and subsequently graduate 95% of all matriculated underrepresented MORE Scholars within six (6) years of enrollment in a doctoral program. Imbedded in these goals are objectives and strategies that provide a specific educational program and a three-tier mentorship process for all MORE Scholars designed to overcome barriers to success. Key evaluation performance measures will be used to determine program effectiveness through a comprehensive evaluation process that includes three distinct entities: the Office of Outreach Programs (manages day-to-day concerns), the MORE Progress Committee (reviews student progress each semester) and the MORE Program Evaluation Committee (yearly reviews the extent the program outcomes and process objectives were met). Our comprehensive program with a network of partner institutions and past success in the recruitment of underrepresented minorities into graduate programs in the biomedical sciences should provide the formula for success with the proposed MORE program. The institutional commitment to these goals is reflected in the participation of the Executive Vice President for Academic Affairs and Research, as well as the Dean and Associate Deans of the graduate school and the level of financial support toward MORE program goals that has been included by the institution. With the foundation of our K-12 and undergraduate programs as providing the training ground for graduate enrollment, the attainment of the proposed goals will be achieved. Public health relevance: There are two major goals of this project: 1) Increase the number of underrepresented minority students entering graduate programs at UNTHSC from undergraduate programs to approach that of the Texas demographic profile of these groups; and 2) Retain and subsequently graduate 95% of all matriculated underrepresented MORE Scholars within six (6) years of enrollment in a doctoral program.

DESCRIPTION: Glucocorticoid (GC) administration has been associated with an increase in intraocular pressure (IOP), particularly in patients with primary open-angle glaucoma (POAG) where greater than 90% are considered glucocorticoid responders as compared to 33% of the general population. The reason for this difference in responsiveness among the population is not clear and the molecular mechanism for the hyper-responsiveness among glaucoma patients still remains unknown. It has been reported that the GC receptor beta (GRbeta), primarily located in the nucleus, functions as a dominant negative regulator of the transcriptional activity of GRalpha, possibly forming inactive heterodimers. Such an action is consistent with recent reports that elevated GRbeta is associated with glucocorticoid resistance in asthma patients. Preliminary observations in our laboratory shows that normal trabecular meshwork (TM) cell lines express relatively higher amounts of GRbeta than glaucomatous TM cell lines and that GRbeta is highly concentrated within the nucleus in these normal but not glaucomatous TM cell lines. This suggests a potential role of GRbeta in contributing to glucocorticoid responsiveness in TM cells. It is proposed that decreased nuclear amounts of GRbeta could result in the enhanced transcriptional activity of GRalpha and contribute to glucocorticoid hyper-responsiveness seen in glaucoma patients. Consistent with this hypothesis has been the finding that the peripheral vascular response to GCs is enhanced in patients with POAG. The overall goal of this proposal is to demonstrate that GRbeta is a key component in the regulation of the GC GRalpha response in ocular tissues and the relative expression of GRbeta is important for GC action. The hypothesis to test is that high expression of GRbeta in TM cells leads to GC resistance, and that low levels of GRbeta expression, as found in glaucomatous TM, leads to GC hyper-responsiveness and ocular hypertension. Four aims are designed to address this hypothesis and include: 1) determine the mRNA, protein expression and subcellular distribution of GC receptors, GRalpha and GRbeta, in normal and glaucomatous TM cell lines and intact tissue; 2) determine if GRbeta expression and overexpression inhibits GC-induced gene expression in TM cells and the increase in GC-induced IOP in isolated perfusion cultured human anterior segments; does GRbeta knockout with siRNA induce hyper responsiveness in normal cells; 3) identify the trafficking mechanism responsible for moving GRbeta from the cytoplasm to the nucleus and determine the role trafficking may have on the accumulation of nuclear GRbeta in normal and glaucomatous TM cells; 4) determine whether polymorphisms in GRbeta splice sites or in the splicesome factor gene SRp30c are associated with glaucoma or with GC-induced ocular hypertension and if in these patients there is a generalized sensitivity to cutaneous vasoconstriction. These specific aims are designed to determine if GRbeta regulates GC responsiveness and if decreases in its expression contribute to ocular hypertension often seen in patients with POAG.

RESEARCH & RELATED Other Project Information 6. Project Summary/Abstract This is a competitive renewal of the Short-Term Research Training for Increasing Diversity in Health-Related Research (R25) that we entitled, ¿Summer Multicultural Advanced Research Training (SMART)¿. This proposal continues a highly successful effort to encourage and recruit underrepresented and disadvantaged undergraduate students to pursue careers in the biomedical sciences by providing a ten week summer traineeship in a research environment that focuses on health disparities and chronic diseases that affect minority populations (e.g. cardiovascular disease, hypertension, obesity, diabetes, stroke, Alzheimer's disease and glaucoma). This program involves a multidisciplinary approach incorporating faculty from among the biomedical science division of the health science center. Trainees attend workshops and lectures on: Responsible Conduct of Research including issues on scientific misconduct, plagiarism, laboratory safety and practice, Animal handling and Animal Rights. Students will take a class on ¿Introduction to Cardiovascular Science¿ and receive two hours of credit toward their degrees at their home institutions. Each student selects a research mentor from the participating faculty after reviewing their research programs and listening to faculty presentations. Students and faculty design and plan a research project that can be accomplished within the ten-week period. All students must complete an abstract suitable for submission and presentation at the Annual Biomedical Research Conference for Minority Students (ABRCMS) that is held each Fall. Students gain experience in: 1) generating a hypothesis; 2) testing their hypothesis through research experimentation; 3) using new research methodology; 4) evaluating and analyzing data using computer techniques, and 5) presenting their findings in a written abstract and oral presentation at the end of the training period. The overall effect of these activities is to create a sense of becoming a member of the scientific research community. This sense is furthered by student participation in their mentor's research group and departmental weekly seminar meetings. A Role Model Seminar is also presented as a part of this summer experience. Students and faculty mentors each evaluate the program and each other at the end of the program. An external evaluator is used to help assess the program process as well as program outcomes. Students will be tracked throughout their participation in the program and after by formal mailed questionnaires, email follow- ups and the use of a list serve. Students will be recruited to return to the McNair summer program, if still eligible, and for entry into graduate school. Program success will ultimately be measured by the numbers of students who enter biomedical science graduate programs and research as a career. RESEARCH & RELATED Other Project Information 7. Project Narrative This program entitled, Summer Multicultural Advanced Research Training or SMART is part of an overall effort by our institution to increase the numbers of underrepresented and disadvantaged students into masters and doctoral programs in biomedical sciences. It is a ten week summer research training experience with specific emphasis on research career development in cardiovascular and related disorders. They will be in research laboratories most of their time, attend workshops on responsible conduct of research and attend a weekly course on cardiovascular science.

DESCRIPTION (provided by applicant): Glaucoma is a leading cause of visual impairment and blindness in the US and worldwide. However, the molecular mechanisms responsible for the development of glaucoma are poorly understood. The use of anti-inflammatory glucocorticoids (GCs) can cause elevated intraocular pressure (IOP) and glaucoma in susceptible individuals. These steroid responders are at higher risk for developing primary open-angle glaucoma (POAG). Almost all POAG patients are steroid responders. GC-induced ocular hypertension and glaucoma is clinically similar to POAG, and GC effects on the trabecular meshwork (TM), the tissue responsible for regulating aqueous humor outflow (and therefore IOP), mimic many of the molecular and biochemical changes that occur in POAG. Two major isoforms of the glucocorticoid receptor (GR) mediate GC biological responsiveness. GRa is a ligand activated transcription factor and GRb is a dominant negative regulator of GC activities. We recently found that glaucomatous TM (GTM) cells have very low levels of GRb compared to normal TM (NTM) cells, which makes GTM cells much more susceptible to the glaucomatous effects of GCs. Our overall hypothesis is that: (a) high expression of GR?; in normal TM cells leads to GC resistance, and low levels of GR?; expression, as found in GTM, leads to enhanced GC responsiveness and elevated IOP in glaucoma and (b) GR?; activity and expression are regulated by specific splicesome proteins and by translocation to the nucleus via specific importins. The following specific aims will address and test this hypothesis: (1) to investigate whether altered expression and/or activity of splicesome proteins regulate GR?; expression in the TM in the context of glaucoma, (2) to delineate the role of differential trafficking of GR?; and GR?; in regulating GC sensitivity in the normal and glaucomatous TM, and (3) to evaluate whether GC-induced ocular hypertension is regulated by GR?; expression in ex vivo perfusion cultured anterior segments as well as in vivo in mice. This research explores for the first time the roles of alternative splicing and nuclear translocation in the TM, will develop 2 new models of GC-induced ocular hypertension as well as will provide better insights into the pathogenesis of GC-induced ocular hypertension, steroid glaucoma, and POAG.

DESCRIPTION (provided by applicant): The Summer Multicultural Advanced Research Training or SMART program at The University of North Texas Health Science Center (UNTHSC) has existed since 1993 as part of its Coordinated Plan to Increase Diversity in the Biomedical Sciences. The program provides underrepresented minorities and individuals from disadvantaged backgrounds and with disabilities a 10-week summer research experience. SMART program trainees are recruited from our existing URM partner institutions as well as through attendance at undergraduate student science conferences. Prospective undergraduate applicants entering their sophomore or junior year are selected based on a GPA of 3.0 or greater and their indication of interest in pursuing a doctoral degree in biomedical sciences. SMART trainees participate in mentored research provided by UNTHSC faculty conducting funded research. Trainees also attend academic courses, study skill and survival workshops that expose and prepare them for entry and matriculation in graduate school. Trainees are provided stipend and room and board through this funding mechanism. To culminate the completion of their summer program, SMART trainees present their research in the form of a research poster competition followed by a Role Model Awards Banquet. In addition, SMART scholars are encouraged to submit an abstract of their research to be presented at a national on conference (SACNAS or ABCRMS). SMART trainees are also encouraged to apply for another year of summer research through the UNTHSC McNAIR Post-baccalaureate program as incoming juniors or seniors. In total, the expectation of this program is that 100% will complete their undergraduate degree and 80% will continue in doctoral degree programs. Thus, increasing the number and diversity of biomedical scientists. (End of Abstract)