This application proposes to develop a Stem Cell Core Facility of ~1700 square feet to support the use of human embryonic stem cells (hESC) for a growing consortium of stem cell scientists at the home institution as well as neighboring institutions. The facility will be built and managed so as to allow use of non-NIH-approved hESC cell lines as well as research funded by non-federal agencies including the California Institute for Regenerative Medicine (CIRM). The Facility will be centrally located adjacent to other existing, successful core facilities and within short walking distance of all the users at the home institution. The Facility will be managed by an Oversight Committee consisting of faculty experienced in hESC and associated technologies, as well as those with experience in managing shared core facilities. The Committee will have close contact with an established Biotechnology Impacts Center to address any ethical issues that may arise.
The users at the home institution consist of an energetic, interdisciplinary group of both young and established investigators who have made a substantial commitment to stem cell biology. Within the past several years, they have held workshops on embryonic stem cells with neighboring institutions, taught two graduate level courses in stem cell biology, including one in bioethics, established a Stem Cell Center, and applied for and received CIRM funding. They have recently hired an experienced hESC investigator and are currently recruiting others, demonstrating the home institution’s commitment to the field of hESC. The group currently consists of 30 investigators from three different colleges within the home institution who have common interests in molecular mechanisms of pluripotency and differentiation of hESC.
Several investigators have joint projects, including collaborations with investigators at neighboring institutions who will also be using the facility. The proposed Stem Cell Core Facility will allow this dynamic group of accomplished investigators to bring the promise of stem cell biology to an expanding, culturally diverse region of California.
The research programs that would use the facility concentrate on various aspects of the molecular mechanisms underpinning the pluripotency of hESC, as well as their ability to differentiate into different types of tissues. The results generated by these programs will contribute to the development of tools, diagnostics, and therapies by laying the foundation for understanding hESC and identifying new compounds and methodologies that will allow researchers to maintain hESC and prepare them for use in therapies. This basic understanding of the molecular networks governing hESC biology is essential before any safe and effective treatment can be considered for use in humans.
When Californians resoundingly passed Prop 71, they demonstrated the importance of stem cell research to all the citizens of our state. However, the human embryonic stem cell (hESC) lines that are currently sanctioned by the federal government are limited by many factors including genetic stability, contamination, poor growth characteristics, and lack of genetic and disease diversity. Working with non-federally approved hESC lines, including new more robust lines that will be developed in the future, will be necessary for any eventual therapeutic use of stem cells. Also critical to that success will be a thorough understanding of the molecular mechanisms that govern the pluripotency and differentiation of hESC, as well as attracting new scientific expertise to the field of stem cell biology.
This proposal meets these challenges and benefits all Californians by establishing a Stem Cell Core Facility (SCCF) that will greatly expand both the scientific as well as the geographic base of stem cell research. The SCCF will allow research on non-federally funded hESC lines and service a group of highly accomplished investigators at the host and neighboring institutions in the most ethnically and culturally diverse and fastest growing region of California. The investigators are all at the top of their respective fields, have a range of hESC expertise and are committed to applying their experience to some of the most critical issues facing the hESC field today. The group is highly interdisciplinary and has an established history of productive interactions and collaborations. They have created a new Stem Cell Center which is aggressively fostering stem cell research and have secured extramural funding for that research. All proposed users have existing projects that directly impact our understanding of the basic biology of hESC and will generate data that will be essential to the successful development of stem cell-based therapies.
SYNOPSIS OF PROPOSAL: This application from Dr. Prudence Talbot is for 3 years of funding to establish a Stem Cell Core Facility (SCCF) at UC Riverside (UCR). She proposes a 1700 sq ft core that will be shared by multiple investigators. The laboratory will consist of one cell culture lab for hESC, one cell culture lab for non-hESC, a separate room for growing feeder cells, plus X-ray irradiator, qPCR, FACS, confocal, karyotyping, and cryopreservation facilities. The lab will be located in space contiguous with the Institute of Integrated Genome Biology. It is predicted that the UCR SCCF will provide services for 15 investigators, 12 from Riverside, two from Loma Linda and one from UC Irvine (UCI).
QUALITY AND IMPACT OF THE SCIENCE: The proposal is responsive the RFA, and the majority of the work will be on non-NIH-approved cell lines. This is a well-organized proposal from Dr. Talbot, a senior investigator who leads this proposal. She has brought together a large number of investigators from UCR, including Dr. Sato, a young stem cell biologist who has been recruited to UCR, as well as faculty from neighboring institutions (such as Loma Linda and UCI) who could utilize this core. Some of the investigators already have been using hESC or have been approved for SEED funds from the CIRM, and one has applied for a CIRM Comprehensive Research grant. All of these investigators will examine basic functions of hESC including the molecular mechanisms that maintain stem cell pluripotency. The variety of the research to be conducted is impressive, including such diverse areas as voltage-dependent channels during differentiation, high throughput assays for pluripotency genes, and the role of non-coding RNAs in hESC. Part of the research strength lies in small molecule screens that may lead to the discovery of compounds for maintenance of undifferentiated hESC in defined media. In particular, Dr. Pirrung has been approved for a CIRM SEED grant in this area of high-throughput screening. Dr. Sauer is an outstanding molecular biologist focusing on miRNAs in Drosophila, and more recently hESC, who also has been approved for a CIRM SEED grant. Dr. Sato will be the driving force for analyzing molecular heterogeneity of pluripotential hESC. Other faculty will be pursuing applications of hESC in the context of skin repair, liver regeneration, brain and spinal cord injury. Collaboration with faculty at Loma Linda University will focus on in vivo localization of ferromagnetically labeled transplanted stem cells. The group in general has a very strong track-record of funding and publications, indicating they are productive scientists, but the program could be strengthened significantly with more developmental biologists. With the exception of Dr. Donovan from UCI, no one has extensive experience with hESC, and thus no one has a true sense of the difficulty of working with hESC. Many have proposed projects that will compete directly with many of the best labs in the business, who have already been doing these projects for years (for example, small-molecule screening on hESC). As a result, these investigators are at a disadvantage compared to other institutions.
APPROPRIATENESS OF SPACE AND EQUIPMENT TO SCOPE OF PLAN: There is a clear need for this facility. There are multiple investigators who would like to work with hESC, some of whom are CIRM funded, but only Dr. Sato currently has space due to his recent recruitment. The proposed space for the shared research lab is appropriate and will benefit from proximity to other core facilities in genomics, proteomics, bioinformatics and imaging. The Program Director has acquired 1700 sq feet of space for the facility in a building that already houses substantial shared equipment. The equipment requested is all reasonable and necessary. Funds are also requested for large equipment for shared facilities including an X-ray irradiator, qPCR machine, FACS, confocal, karyotyping, and cryopreservation facilities. The facility will have one hESC culture room with two biosafety cabinets, one cell culture room for non hESC culture with one biosafety cabinet, and one feeder culture room with one biosafety cabinet. Three dissecting scopes are requested, but it is not clear why since only two people can be working on hESC culture at any one time. Overall, the organization of the space is well described, but the facility is relatively small and the amount of tissue culture space seems limited. hESC are extremely time-consuming to culture, so four hoods and only four incubators seems conservative, especially when contamination can be problematic and the investigators will want their own incubators, and sometimes separate incubators for MEF expansion.
QUALITY OF MANAGEMENT PLAN: Dr. Talbot will serve as lab director and is the interim Director of the Stem Cell Center. Appropriate UCR faculty will provide oversight particularly Dr. Sato and Dr. Pirrung. The plan for administration of the lab with appropriate oversight is excellent. A key interaction involves coordination with Dr. Donovan who is co-director the UCI stem cell center. Dr. Donovan runs a highly recognized program in stem cell biology. In fact, more information on the advisory and management role of Dr. Donovan would strengthen the application since coordination of stem cell activities on the two campuses is an asset in the long term. The laboratory itself will be overseen by 100% of an experienced lab manager, 20% of a FACS technician, and 5% each for a technician for FACS, genomics, and cytogenetics. These people are not yet in place, and the lack of extensive experience with hESC from any investigator intimately involved with this facility is a real concern. Because the day-to-day management of the facility will be done by a PhD coordinator, and because of the difficulty of working with hESC, it is essential that a highly experienced individual, familiar with hESC, is hired into this position. It is hard to believe that such a person can be hired for ~$50K, and that the technologist supporting this position (with an MA/MS) can be hired for $35K. hESC are extremely difficult to work with and the entire facility, from personnel on down to the type of hoods, has to be designed and selected by someone who really knows the cells. Provided that highly experienced people can be found and hired for the proposed salaries, Dr. Talbot may be able to establish a high quality facility for hESC culture that is badly needed at UCR. However, this team still has a lot to learn regarding establishing and running a shared hESC facility. For example, one technician will not be sufficient for the number of planned users. Moreover, there are concerns with regard to the operation and maintenance of some of the equipment. The facility will contain a BD FACS Aria, but no analyzer. The Aria is not really well designed for analysis, only for sorting, so an analyzer that is nearby is essential to monitor marker expression on the hESC as a first-pass for pluripotency. An operator who works at a different facility will oversee this Aria, but this is not really practical- the Aria, despite its advertising, does not work well as a relatively unsupervised instrument. Likewise, it may be better to send the karyotyping work out to an experienced lab as hESC karyotypes can be more difficult than standard karyotypes and the volume here does not necessarily justify a karyotyping system.
DISCUSSION: This shared lab is part of the recently organized stem cell biology center at UCR headed by Dr. Talbot. A number of faculty are interested in the applied (translational) aspects of stem cell research, yet this group was unable to write a strong application because they don’t yet know the pitfalls of setting up an hESC core. A key issue is the interaction with Dr. Donovan at UCI. The application would have been strengthened if the advisory role of Dr. Donavan and the coordination of activities between the two campuses had been more fully developed. The proposed program has a faculty grouping that is considered excellent, and there is a good effort here to build a stem cell program, yet reviewers believe the application reflects a lack of experience in what is required to run an hESC shared lab. The principals involved lack experience with hESC and given the difficulty of working with these cells this is a real concern. A lack of hESC experience is also represented by some disconnect in the lab set up and the required equipment. The applicants do not have the resident expertise and need to hire it in, but they have under-budgeted for finding people with the right expertise. Also, they do not address security and access very well. Altogether, these issues seem to reflect a lack of understanding. One discussant noted that while the salaries proposed for staff to run the facility are likely to be inadequate, one can’t directly compare salaries at institutions in different parts of the state given that there are differences in the cost of living. Another discussant noted that this is an institution with an emerging program showing a lot of academic promise that has a real need for this type of facility. Concerns were reiterated about the lack of management and personnel with hESC experience, despite already having an ESCRO committee, some security measures, and Dr. Sato on board for expertise.
PROGRAMMATIC REVIEW: A motion was made to recommend this Shared Research Laboratory application not be funded. While this institution has talented faculty and there are people interested in the facility, they currently lack a critical mass of research expertise. The shared lab also lacks experienced staff and has a weak structure. This application could be improved, but it is not an easy fix with the number of flaws and the lack of expertise. The question was raised of whether it could be improved if they found the expertise. The response was no, they would have to fully redesign and reallocate resources, thus this seems more problematic to fix than by simply bringing in a more experienced staff. Another question was why couldn’t UCR use the UCI facility? The director of the UCI facility is on the UCR team, but not taken advantage of well enough in this application. Another panel member pointed out that there were some fatal flaws, and that this proposal was not quite within the first quartile. The motion to recommend that this Shared Research Laboratory application not be funded passed.