This CAREER proposal applies engineering approaches to generate fundamental knowledge describing how mitochondria function within intact mammalian cells. Mitochondria serve as both the powerhouse and chemical processing plants in virtually all higher organisms on earth, supplying the energy and building blocks necessary for life. Defects in mitochondria contribute to numerous diseases that include diabetes, cancer, neurodegenerative diseases, and aging. Despite their central importance in energy metabolism and physiology, several aspects of mitochondrial function remain unknown. For example, we still do not know how many important nutrients and chemicals enter and exit the mitochondria where they are broken down or synthesized. Control of such processes using drugs could treat numerous diseases. This proposal will address such questions by engineering and modeling advanced cell systems to study mitochondrial metabolism. This research will generate fundamental knowledge that can be used to improve human health, disease outcomes, drug development, and nutrition. The metabolic physiology research will also be integrated with an educational program that stimulates student and educator interest in biochemical engineering, biochemistry, and medicine. To accomplish this goal development of a lab-based program with a local high school is proposed to expose students to bioengineering research and its application to human physiology, relating yeast metabolism to human diseases such as cancer and diabetes. In addition a web-based interactive tool will be developed to enhance the ability of higher level students and researchers to learn biochemistry and metabolism.
It is proposed to identify new targets for selectively controlling substrate utilization in eukaryotes in the context of human disease and metabolic engineering applications. The interplay between mitochondria and eukaryotic cells is one of the most important symbiotic relationships on earth. Despite the extensive studies on mitochondrial structure and metabolic activity conducted over the last century, the identity and function of numerous mitochondrial components remains a mystery. Stable isotope tracing and systems-based metabolic flux analysis (MFA) provide the most effective means of quantifying cellular physiology in lower organisms, but application to more complex species is complicated by compartmentalization. The investigator developed a new approach for tracing metabolic pathways in specific subcellular compartments such as the mitochondria and cytosol. In the first objective within the proposed project this technique will be exploited to develop a comprehensive, compartmentalized model of eukaryotic metabolism and cellular redox potential. Cellular engineering strategies will be applied to functionally annotate the genes/proteins that transport metabolites into and out of mitochondria. These engineered cells will ultimately be characterized using the model developed in the first objective. These results will significantly advance the functional understanding of eukaryotic cell metabolism and improve the ability of clinicians and engineers to control mitochondrial function. This research program will be integrated into a broader educational theme that aims to educate students, teachers, and the public on biochemistry and metabolism. This educational plan includes efforts to (i) highlight the impact of STEM fields and in particular engineering approaches on human health, industry, and the environment; (ii) initiate a lab-based program with a local high school to expose students to bioengineering research and its relevance to human physiology; (iii) develop a web-based learning tool as part of our senior design course to promote and accelerate the learning of metabolic pathway interconnectivity by students and researchers. These research and education activities will also provide training opportunities for undergraduate, graduate, and postdoctoral scholars.
This CAREER award by the Biotechnology and Biochemical Engineering Program of the CBET Division is co-funded by the Systems and Synthetic Biology Program of the Division of Molecular and Cellular Biology.
