Grant Award Details
Explore a hypothesis that mitochondria both reflect and influence hESC decisions to self-renew versus differentiate
Grant Application Details
- Role of Mitochondria in Self-Renewal Versus Differentiation of Human Embryonic Stem Cells
Human embryonic stem cells (hESCs) hold great potential for treating multiple human dread diseases, including but not limited to cancer, diabetes, obesity, Alzheimer disease, and certain types of heart failure. However, a growing appreciation exists for the notion that not all hESCs have identical capabilities in correcting or ameliorating disease and not all hESCs will be valuable as potential therapeutic cell sources. Because hESCs contain genetic information like all human cells, some hESCs will have genetic mutations or alterations that will make them more or less desirable for therapy. The heritable information contained with hESCs comes from DNA in the cell nucleus and also from DNA within maternally inherited mitochondria. In fact, it is the functional capabilities of mitochondria in hESCs that this proposal addresses because over 400 mutations in mitochondrial DNA result in disease and many more disorders associated with mitochondrial dysfunction, often unidentified at the molecular level, arise from mutations in nuclear DNA.
It is potentially dangerous that so little is known about the functional capabilities and role for mitochondria in hESCs and in the major decisions that hESCs make, such as whether to self-renew and make more stem cells or to differentiate into any one of the known human lineages, including muscle, skin, brain, and other cell types. We anticipate the day when stem cell therapies to combat disease or provide replacements for worn out components will be a main part of individualized medical treatment. We believe it is therefore critical to choose the best stem cell starting materials for such therapeutic applications. This view, combined with a desire to understand how mitochondria, as the main source for a cell’s energy and building block generation, functions in stem cell decisions, propels us to provide 3 integrated areas of specific investigation into stem cell mitochondria and their role(s) in decision making. Our studies will evaluate basic mitochondrial functions and structures in a variety of hESCs to gain an appreciation for variability in distinct hESCs (Aim 1). We will alter mitochondrial function in hESCs with genetic and environmental insults to gain an appreciation for the global effects on hESC function and as a way to help select appropriate stem cells for future therapeutic applications (Aim 2). We will force hESCs with normal or altered mitochondria to differentiate into germ cells, blood cells, skin cells, or brain cells with expert collaborators in each lineage type to evaluate how the state of mitochondrial function will dictate the ability for hESCs to provide multiple, distinct replacement lineages for use (Aim 3).
In sum, we expect our studies will reveal the critical role for mitochondria In stem cell biology and this new knowledge and our analytical approach will help provide essential information for choosing optimum stem cells for future therapeutic applications.
Our proposal will benefit California by adding new knowledge on the functional capabilities of human embryonic stem cells (hESCs) and their lineage differentiated derivatives, which will support the California peoples’ and taxpayers’ commitments to individualized medical treatments of the near future. It will help us select the best possible stem cells for study and therapy development in our major academic centers and will provide information to many of California’s biotechnology and pharmaceutical companies in the burgeoning stem cell industry, whose success will propel hiring and increased economic prosperity for the state. Our work will provide additional information to patient advocates, ethicists, and even (eventually) medical geneticists to help select the optimal course for developing and modifying stem cell usage policies and infrastructure within California. This proposal will provide information for patients and their physicians, that may, at some future time, impact the selection of particular stem cell attributes for specific types of therapeutic applications. In sum, the added knowledge provided by a detailed analysis of mitochondrial capabilities in hESCs will have tangible health and economic impact on California, its academic institutions and biotechnology/pharmaceutical companies, and the rest of the nation as California and its people move forward with personalized medicine during the 21st century.
- Stem Cell Reports (2014) Defining the role of oxygen tension in human neural progenitor fate. (PubMed: 25418722)
- Nat Methods (2014) Live-cell mass profiling: an emerging approach in quantitative biophysics. (PubMed: 25423019)
- Methods Enzymol (2014) Techniques to monitor glycolysis. (PubMed: 24862262)
- Dev Cell (2013) A small molecule inhibitor of redox-regulated protein translocation into mitochondria. (PubMed: 23597483)
- Biophys J (2013) Quantification of biomass and cell motion in human pluripotent stem cell colonies. (PubMed: 23931307)
- Proc Natl Acad Sci U S A (2012) Correcting human mitochondrial mutations with targeted RNA import. (PubMed: 22411789)
- Nat Protoc (2012) Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells. (PubMed: 22576106)
- Cell Stem Cell (2012) Metabolic Regulation in Pluripotent Stem Cells during Reprogramming and Self-Renewal. (PubMed: 23122286)
- Stem Cells (2011) Mitochondrial function controls proliferation and early differentiation potential of embryonic stem cells. (PubMed: 21425411)
- EMBO J (2011) UCP2 regulates energy metabolism and differentiation potential of human pluripotent stem cells. (PubMed: 22085932)
- Lab Chip (2010) Microfluidic image cytometry for quantitative single-cell profiling of human pluripotent stem cells in chemically defined conditions. (PubMed: 20390128)
- Cell (2010) PNPASE regulates RNA import into mitochondria. (PubMed: 20691904)
- Cell Stem Cell (2009) Induced pluripotent stem cells and embryonic stem cells are distinguished by gene expression signatures. (PubMed: 19570518)
- Stem Cells (2009) Derivation of primordial germ cells from human embryonic and induced pluripotent stem cells is significantly improved by coculture with human fetal gonadal cells. (PubMed: 19350678)
- Stem Cells (2009) A self-renewal program controls the expansion of genetically unstable cancer stem cells in pluripotent stem cell-derived tumors. (PubMed: 19224508)
- J Vis Exp (2008) Probing for mitochondrial complex activity in human embryonic stem cells. (PubMed: 19066553)
- J Vis Exp (2008) From MEFs to Matrigel 2: splitting hESCs from MEFs onto Matrigel. (PubMed: 19066543)
- Nature Reports Stem Cells (2008) How to Assess a Stem Cell Genome
- J Vis Exp (2008) From MEFs to matrigel I: passaging hESCs in the presence of MEFs. (PubMed: 19066554)
- J Vis Exp (2008) From MEFs to Matrigel 3: passaging hESCs from Matrigel onto Matrigel. (PubMed: 19066542)
- Stem Cells (2008) Copy number variant analysis of human embryonic stem cells. (PubMed: 18369100)
- Cancer Cell (2007) Enhanced paracrine FGF10 expression promotes formation of multifocal prostate adenocarcinoma and an increase in epithelial androgen receptor. (PubMed: 18068633)