Development of high-throughput screening platform for directing hESCs into pancreatic progenitor cells and mature pancreatic beta cells

Funding Type: 
New Faculty II
Grant Number: 
ICOC Funds Committed: 
Public Abstract: 

Human embryonic stem cells (hESCs) have been recognized as an inexhaustible source for regenerative medicine, a promising platform for developing and testing new drugs as well as an invaluable tool to study human development. However, the mechanisms about how hESCs maintain its multiple lineage development potential (also known as pluripotency) and differentiate into different adult cell types in responding to exogenous stimuli are far from clear. The ability to direct controlled differentiations of hESCsinto specific cell types, e.g. pancreatic-cells, will significantly advance the hESCs research and translation into medical and pharmaceutical applications.
We propose to develop critical basic tools of establishing three-dimensional hydrogel-based cultural technique. We will then use this technology platform as cell-based high-throughput assays for hESCs directly differentiation into pancreatic progenitors and pancreatic ÔÅ¢-cells. We will use multiple reporter genes at the same time for each lineage/stage differentiation in order to ensure the fully differentiation program in responding to stimuli. We will first generate multiple reporter gene constructscontaining the EGFP/ECFP/ERFP, driven by multiple promoters of established pluripotency and differentiation genes. We will then establish hESCs clones stably expressing these reporter genes and culture these engineered hESCs in 3-D hydrogel environment in order to develop into high throughput screening platform. To develop cell-based high-throughput assays for detecting directed hESCs differentiation, we will test these reporter cell lines in 3-D culture environment by applying established multiple step dfferentiation procedure. We will then perform high throughput screening for hESCs differentiations into pancreatic progenitors, as well mature pancreatic ÔÅ¢-cells. We will exam the expression of endogenous lineage differentiation markers of hESCs by immunofluroescence staining. Finally, we will elucidate the targets of screening hits by combinatorial approaches to elucidate the underlining mechanisms. The host university is a center of excellence for chemical genomics with CIRM-funded stem cell faciliy and a NSF-supported chemical genomics platform, and committed to support stem cell research.
Technologies to make the stem cell manipulation easier in culture and to more efficiently direct their maturation into adult cell types are vital to further developments in the field. Stem cell technology provides a paradise of cell research, where rapid advances will make immediate impact on cell replacement therapy and drug discovery/development. This proposal will lead to new technologies and products developments and the outcomes of the project will enhance our capability of manipulating hESCs for medcal and pharmaceutical applications and particularly facilitate the eventually development of treatments for degenerate diseases like diabetes.

Statement of Benefit to California: 

The disability and loss of earning power and personal ability resulting from a disease or disorder are devastating and create a financial burden for California in addition to the suffering caused to patients and their families. Although it is only in its early stage, human embryonic stem cells (hESCs) have already been shown to be capable of long-term self-renewal in culture and have remarkable potential to develop into many different cell types in the body. They therefore represent an infinite source of precursor cells to treat degenerative, genetic diseases, or injury, and trauma. Technology and products resulting from this project will enhance our capability of manipulating hESCs for pharmaceutical applications and particularly facilitate the eventual development of treatments for degenerate diseases like diabetes. Improved function in patients afflicted with these diseases will greatly promote the public health and result in tremendous savings to California in healthcare costs, particularly in the areas of long-term care. Federal constraints on stem cell research create a critical need for non-federal funds to achieve these goals. Funding by the California Institute for Regenerative Medicine improve California’s stem cell infrastructure and speed the translation of basic university research into medical products that change lives.

The improved ability to maintain viable stems cells as individual cell in culture and to better control their maturation by exogenous stimuli will unlock whole new techniques for working with these valuable but sophisticated resources. The proposed project, when successful and scaled up, eventually will serve to improve the source, storage, price, availability, quality, purity, and diversity of a wide variety of stem cell types as used in many downstream medical, biotechnology and pharmaceutical projects. A ready source of conveniently available stem cells of all types, in turn, will act as a catalyst for new fundamental discoveries in stem cell biology, and will provide enabling reagents for many future products and methods. These discoveries, products, and methods will improve the tax base, create many new jobs, and save billions in healthcare costs in California.

The 3-D culture, high throughput screening technologies and small molecular modulator(s) produced in this program will have broad implications for the advancement of medical and life science related to human stem cells, including replacement tissue treatments and drug discovery. Stem cell technology is currently a very strong research area where rapid advances are possible and the research in developing new products and unlocking basic understanding of how human cells function is of very high value. California is viewed as a world leader in stem cell research, biotechnology and pharmaceutical research and development, and the advances in the field made here will contribute to the California’s leading position in these industries immediately.