Characterization of the regulatory events that lead to changes in gene expression and function is key to understanding the molecular mechanisms by which biological processes are governed. To address questions aimed at defining these regulatory interactions, it is necessary to accurately quantify changes in gene expression that result in downstream effects on cell/organism function. To that end, real-time Polymerase Chain Reaction (PCR) analysis provides a method by which changes in gene expression can be quantified. This approach allows precise assessment of transcriptional targets whose gene products are responsible for modulating adaptive responses. This grant will support the purchase of a Polymerase Chain Reaction machine that will be enable real-time PCR methodology to investigate multiple projects centered on examining diverse biological processes including immunology, cell survival, inflammation, tissue repair, differentiation, and smooth muscle cell biology. Projects of note include characterization of the effects of mechanical forces and extracellular matrix interactions on modulating function of smooth muscle cells and differentiation in mesenchymal stem cells. In the area of inflammation, the processes that regulate inflammatory responses in immune cells and brain as well as the downstream transcriptional targets of immune modulators will be investigated in macrophage and glial cell lines. These studies will be aimed at understanding the molecular mechanisms that result in initiation of cell survival and tissue repair in glial and neuronal cells. Finally, this technique will also be applied to study the role of the unfolded protein response during immune response, with a focus on cytokine production by B lymphocytes.
Assessment of gene expression patterns will allow the faculty at Mt. St. Mary's University to make more significant contributions to their research fields and will directly impact the training of students by enhancing the undergraduate educational experience. The current proposal describes a total of 11 research and educational projects to be carried out by four faculty members. Importantly, several of the projects address approaches for incorporation of real-time PCR methodology into both the undergraduate curriculum and outreach educational programs aimed at local high schools. This instrumentation provides a means by which to incorporate an inquiry based approach to science instruction. Rapid advances in the understanding of cellular and biochemical processes as a result of genetic and genomic research have generated considerable demand for individuals trained in biotechnology, genetics, biochemistry, and cell biology. Funds from this grant will be used to meet the challenge of improving undergraduate education in STEM areas and enhance student preparation for successful careers in the life sciences.
In 2011, Mount St. Mary’s University (MSMU) was awarded a Major Research Instrumentation award to obtain a quantitative PCR System. This instrument has benefitted the research programs of multiple faculty members within the Science department, the undergraduate biology program, and an outreach program to high school students. One of the research laboratories being served by this project is that of Dr. Dana Ward. A major research focus of the Ward lab is in investigating the mechanisms that regulate vascular smooth muscle cell growth. Abnormal smooth muscle cell growth is a hallmark of many vascular diseases including pulmonary hypertension. Recent work in the Ward lab has focused on examining changes in gene expression in vascular smooth muscle cells isolated from normal patients vs. those with pulmonary hypertension. Using qPCR, Dr. Ward and her undergraduate research students have uncovered several major changes in gene expression in diseased cells. These findings are exciting in that they provide greater insight into the mechanisms that drive disease progression. Another research laboratory served by this project is that of Dr. Katy Dye. The Dye Lab focuses on the effect that virus infection has on cells and the activation of a set of molecular signals called the unfolded protein response. The response is stimulated when the cell’s protein production machinery is overloaded, as is often the case during viral infections. Recent work in the Dye laboratory focuses on determining whether this response is induced during the course of infection by Ebolavirus, a lethal hemorrhagic fever virus that is the focus of several anti-bioterrorism efforts. Dr. Dye and her student have completed pilot studies to optimize the qPCR techniques for her system. Ultimately, their findings may provide rationale for prevention and treatment of Ebolavirus infection, for which there is no FDA-approved cure. Use of this instrument is also vital to the research of Dr. Christine McCauslin. The McCauslin laboratory investigates the mechanisms that regulate inflammation and tissue repair. Initiating inflammation is essential for triggering tissue repair; however, uncontrolled inflammation can result in severe damage to tissues. Understanding these mechanisms is particularly important for understanding the damage and repair mechanisms at work in the brain during stroke. Dr. McCauslin and her undergraduate students have recently used qPCR to characterize gene expression changes during inflammation. Future experiments will utilize qPCR in a high throughput assay to identify specific genes that are important for regulating these processes. Not only do these projects advance the research within their respective fields, they have the dual purpose of undergraduate training. Undergraduate training is the major mission of our university and possessing the pPCR instrument allows us to address this mission in new and exciting ways. It exposes our students to modern instrumentation, helps to cultivate their scientific thinking and prepares them for future careers in STEM areas. In addition to the research pursuits made possible by the acquisition of the qPCR instrument, several educational initiatives have also benefitted. The Science department at MSMU recently redesigned the curriculum of the biology major to include a required Genetics course. With the acquisition of the qPCR instrument, we designed a qPCR laboratory module focused on the topic of inflammation, where students investigate changes in gene expression that result from treatment of immune cells with inflammatory chemicals. Because of the Genetics requirement for the biology major, 100% of our biology majors (excluding cases of transfer students) will now have experience with quantitatively analyzing gene expression. This is an exciting experience for students and a unique educational opportunity at the undergraduate level. We are drafting a manuscript to be submitted to a science education journal, so that we can share our qPCR lab module with other educators. In addition to undergraduate training, we have also employed the qPCR instrument in a program targeted to high school students. MASE (Mount Academy for Scientific Excellence) is a program that provides high school students with hands-on, inquiry based laboratory experience in molecular biology. The MASE curriculum is designed to allow students to ask questions regarding the processes that regulate inflammation at the molecular level and now include an experiment using qPCR. For all of the students, this experience represented their first exposure to qPCR and we believe that it has provided an invaluable training experience that will help prepare them for college-level courses. Obtaining the qPCR instrument has significantly enhanced the research and teaching abilities of faculty and students of MSMU. In the first year alone, we have had nearly 90 undergraduate students perform qPCR experiments either in a classroom setting or in a research laboratory to ask questions about gene expression in different experimental systems. We have additionally served over 20 high school students by enabling them to gain experience with qPCR technology. We look forward to continued implementation and expansion of our research and educational initiatives with this instrument.
