The CIRM Shared Human Embryonic Stem Cell Core Laboratory will provide shared research facilities for use by California scientists. This laboratory will be hosted by a research institution focused on basic research into three of the most important medical problems of modern times: cardiovascular disease, AIDS, and neurodegenerative disorders. Each of these research areas addresses promising targets for regenerative medicine. We propose to develop a laboratory (1108 sq ft) for hESC tissue culture with specialized microscopy, and an animal holding and procedure space (500 sq ft) for in vivo pre-clinical studies of hESCs in mouse models of disease. The proposed laboratory will also help to train students from a nearby college be become laboratory technicians. This facility will contain advanced equipment for analyses of hESCs and complement existing space and incorporate hESC work provided by other core laboratories such as the genomics and flow cytometry cores that serve a broad community of researchers.
The host institution is renowned for the quality and administration of its extensive core facilities. Highly productive cores have always been at the heart of this institution’s culture and this continues to be a priority. Five years ago, the host institution founded an embryonic stem cell core, which allows investigators not familiar with ESC research to obtain training, expertise and knowledge regarding embryoid bodies and ESC differentiation. As a result, two-thirds of the current investigators have incorporated some aspect of stem cell research in their portfolio. The host institution is also located in close proximity to a major biomedical university, so that all stem cell services are being coordinated to provide the best possible array of services to all stem cell investigators.
The research interests of our investigators that are related to stem cells can be grouped into three areas: cardiovascular development and disease, neurodegeneration and repair, and mechanisms that control the genetic stability of the cells while they divide and develop. This research involves the creation of genetically altered ESCs that require maintenance, expansion, and characterization. To aid in the analysis of the cellular phenotypes, we propose to use advanced high-content microscopy equipment. Several leading laboratories that apply this technology to basic cell biological analysis are close to Gladstone. An important next step will be to examine the behavior, survival, and interactions of hESCs once they have been implanted into mice. Visualization of the cells in live animals will be greatly enhanced by the proposed imagining instrument that will allow us to examine living cells within animals by light signals transmitted from the implanted cells. This program represents a comprehensive basic approach to how stem cells develop into other kinds of cells and will form the foundation for future preclinical studies.
Contribution to the California Economy: A major goal of regenerative medicine is to repair damaged tissue. Our research focuses on developing new methods to differentiate human embryonic stem cells (hESCs) into specific cell types for regeneration of diseased tissues. Our research could benefit the California economy by creating jobs in the biomedical industry by developing new technology. Ultimately, this study could help reduce diseases, including cardiovascular, immune, and neurological diseases. Thereby, we hope to increase the productivity and enhance the quality of life for Californians.
The results of our studies will help develop new technology that could contribute to the California biotechnology industry. Our studies will create multiple lines of hESCs that have genetic markers that turn on at specific time points. These cell lines could be valuable for biotechnology companies and researchers who are screening for drug compounds that will cause these developmental changes. Furthermore, we are working closely with California companies to develop new microscopes and analysis software that could be the basis for new product lines or new businesses. If therapies do come to fruition, we anticipate that California medical centers will be leading the way.
The most important contribution of this study will be to improve the health of Californians. Diseases that are the target of regenerative medicine are major causes of mortality and morbidity, resulting in billions of dollars in healthcare costs and lost days at work. As we continue our efforts in medical research, we hope to one day unlock the secrets of tissue development and repair. This knowledge will help medical researchers develop beneficial therapies beyond what is currently available and potentially improve the quality of life and life expectancy of patients who suffer from disease.
SYNOPSIS OF PROPOSAL: The proposal is for the development of an 1100 square foot Shared Research Laboratory (SRL) for tissue culture and specialized microscopy and for a vivarium of approximately 500 square feet for translational model studies. The shared lab will support the great breadth of interest in human embryonic stem cell (hESC) research in the Gladstone Institutes and at the neighboring UCSF Mission Bay campus. The scientific focus for hESC work at the SRL includes the use of hESCs for cardiac and neuron repair and regeneration as well as research into the genomic changes defining pluripotency and differentiation. The institution currently has CIRM Scholars under the training program, several of its investigators have been approved for CIRM Leon J. Thal SEED grants and many of the institution's laboratories are engaged in some aspect of stem cell research.
QUALITY AND IMPACT OF THE SCIENCE: This is a strong proposal from a top-flight institution. The investigators who will use the shared lab are highly qualified and there is a great breadth of interest in hESC research at the Gladstone Institutes and at the neighboring UCSF campus. The use of hESCs fits well into two of the Gladstone’s main research missions, cardiovascular and neurodegenerative disease. The investigators will utilize the shared laboratory space to conduct research on hESCs in three primary areas: a) differentiation of hESCs into mesodermal and cardiac lineages in vitro and the pathways involved in these lineage progressions for treatment of cardiovascular disease; b) generation of neural progenitors from hESCs in which genes involved in neural connectivity and survival are knocked down and their in vitro and in vivo development studied for treatment of neurodegenerative diseases; and c) study of genomic changes underlying pluripotency and differentiation. They will be using state-of-the art approaches to explore important questions in the field including the implementation of bioluminescent/fluorescent imaging to non-invasively follow cell fate and differentiation in mouse model systems. To carry out the above, Dr. Srivastava, the program director(PD), has recruited a group of top-flight investigators at the Gladstone Institute and collaborators at the UCSF and other institutions. Some members of the group have extensive experience with mouse ESCs and Dr. Conklin (associate director) leads the mouse cell core. The PD has an impressive track record in publication and is very well funded as are many other members of the group. A number of investigators have received CIRM SEED grants and the institution also has ten CIRM-funded post-doctoral fellowships under the CIRM training grant. There is no doubt that the proposed studies will generate high quality research that will have high impact. They also plan use the shared lab to develop novel and useful education and outreach ties with the San Francisco Exploratorium and the City College of San Francisco.
APPROPRIATENESS OF SPACE AND EQUIPMENT TO SCOPE OF PLAN: The space available for development of a hESC lab seems modest, if not inadequate (1100 sq. ft) given the number of investigators that will use this resource. The equipment they request is large, but appropriate and justified overall. The personnel named are appropriate and experienced.
QUALITY OF MANAGEMENT PLAN: The management plans for the lab (both overall and on a daily basis) as well the oversight are all excellent. Dr. Srivastava, the PD is outstanding. He and Dr. Conklin will have overall responsibility for running the lab and they have identified individual scientists within the San Francisco area to serve on an oversight committee. They have an experienced cell biologist, Dr. Russnak, who currently runs their ESC facility, who will be responsible for the day-to-day running of the lab. All of this seems fine, but Drs. Srivastava and Conklin only list a 5% effort each and the reviewers expressed concern as to whether this is sufficient. The applications names Dr. Aalto-Setala as one of the key personnel, but the reviewer could not find any description of his/her role in this section.
DISCUSSION: This is a very impressive application from a premier institution. The PD is a very strong investigator as are each of the many faculty who would use the shared space. The PD has many high impact publications and funding. The institution has research strengths in cardiovascular disease, HIV and neurodegenerative diseases. Sixty percent of the investigators at the institution will use hESC or hESC based techniques. There is a major collaboration with UCSF. The scientific focus for hESC work includes the use of hESC for cardiac and neuron repair and regeneration as well as research into the genomic changes defining pluipotency and differentiation. Investigators will be differentiating hESC into mesoderm, labeling with BACs and fluorescent reporters, and doing in-vitro differentiation to cardiac lineages for in-vivo analyses. The effect of gene knockdown in ESC of genes involved in neural connectivity and neuronal survival will be studied in-vitro and in animal models. There is a major emphasis on Alzheimer’s disease through the study apoE and its isoforms in the hippocampus using hESC transplantation. Sophisticated imaging methods will be used in both the cardiac and neurobiology work. There are interested UCSF faculty collaborating in this area – Dr Fisher at UCSF will contribute lines and culture methods. The other major area of research interest in a shared laboratory is in genomic changes (stability/histone regulation/gene regulation) underlying pluripotency and differentiation. The institution has recruited Dr. Yamanaka from Japan who published a seminal paper last year on the minimal requirements for reprogramming of mouse somatic cells to ES-like cells. The applicants also propose that the shared lab will contribute to developing educational outreach programs with the Exploratorium and with City College of SF.
One negative point is that the 1100 sq ft lab space and the one technician proposed will likely be inadequate given the interest, depth and breadth of science planned to be carried out in the facility. The management plan and the oversight committee are well documented and are excellent. The PD is an outstanding scientist and has assembled a well qualified team. The distribution of management between the program director and co-director with complementary expertise is good. The PD has included a long list of letters of support indicative of the breadth of interest in the shared lab. The percentage time commitment is only 5% for each PD and associate PD, this may be insufficient. A discussant noted that the PD has good commitment but only charges the grant for 5% and likely will provide more leadership than what is charged to the grant.
Development of a hESC lab at the Gladstone Institutes seems well warranted given the scientific direction of the Institutes and the number of scientists there who would use the facility. Its close ties with UCSF and significant number of senior and accomplished scientists is important.