High throughput derivation of disease-specific human induced pluripotent cell (hiPSC) lines: use of the [REDACTED] hiPSC selection and characterization platform to derive high quality, fully characterized pluripotent cell lines to support the CIRM iPSC de
Academic, pharmaceutical and biotech research has resulted in the development of incredibly effective medicines and therapies to treat disease. However, testing and developing potential therapies remains inefficient and costly. The process would be greatly improved with a method of accurately representing the diversity of human disease in a format applicable to drug identification. The recent discovery of Induced Pluripotent Stem Cells (iPSC) may allow the modeling of all human disease in a petri-dish. iPSC technology allows the “reprogramming” of any easily accessible cell type e.g. a skin cell or a blood cell, to become any other cell type in the body e.g. a brain cell, a heart cell or a pancreatic cell. Research has already shown the potential of this approach e.g. taking a skin cell from an individual suffering from a genetic disease such as Parkinson’s, reprogramming it to generate iPSCs, expanding these cells and then turning the cells into neurons (the dysfunctional cells responsible for Parkinson’s). In this way, a model of disease can be generated on which drugs can be tested. The current proposal is to use an industrial-scale iPSC technology previously developed in California to generate a stored bank of iPSC cells from thousands of individuals suffering from many different diseases. This bank will be a resource to researchers in universities and pharmaceutical companies who wish to develop models of human disease and identify more effective and safer drugs.
Statement of Benefit to California:
The current proposal is to generate the world’s largest bank of Induced pluripotent Stem Cells (iPSC) from a list of specific genetic diseases. This activity would be directly funded by proceeds from the California Stem Cell Research and Cures Act – Proposition 71 and would fund California-based research and technology organizations. California has consistently led the world in high-tech and biotech innovation. The advent of iPSC technology has the potential to shape the future of drug development and regenerative medicine for the primary benefit of patients. The establishment of the CIRM iPSC bank will maintain California’s position as a center of excellence for biological research and specifically regenerative medicine. Moreover, work to establish and maintain the bank will not only actively employ skilled researchers here in California but will help support established and ongoing Californian research and industrial operations that have been active employers for many years. The effort will also require close collaboration between the Californian research, biotech and technology sectors, potentially leading to other, unanticipated technologies and therapeutic developments.
The applicant proposes to generate 4551 human induced pluripotent stem cell (hiPSC) lines from 1517 individual donors, using a high throughput manual approach. The applicant requested, and CIRM’s president granted, an exception to the eligibility requirement that the project must be a proposal to derive 3 hiPSC lines each from a minimum of 3000 tissue donors. The for-profit applicant institution is located in California. The applicant intends to use skin biopsies as the preferred source of primary cells for derivation of hiPSC, while peripheral blood or other adult somatic cells can be used if necessary. A quarantine process is included to detect and exclude samples with mycoplasma contamination. The genes encoding reprogramming factors will be delivered using a genome integration-free method in combination with small molecule compounds. The characterization of derived hiPSC lines involves gene expression analysis, both at the pluripotent stage and following directed differentiation into cell types representing the 3 germ layers. Verification of chromosomal integrity and of identity between the source cells and the corresponding hiPSC lines will be accomplished by molecular analysis. Loss of the reprogramming vectors from the cells will be confirmed by polymerase chain reaction (PCR). For cell line tracking, the applicant proposes to use an off-the-shelf laboratory information management system (LIMS). Protocols - The protocols are well presented and laid out in a logical manner and the proposed allocation of personnel to specific tasks is well considered. However, a major weakness is that the proposal does not include automation for cell culture. - The proposed hiPSC clone selection process is an interesting approach. However, it is a serial method and thus less scalable than other approaches, posing a limitation on throughput. - The applicant intends to use feeder cells during the selection and characterization process, which will require the end user to use feeder cells or adapt hiPSC lines to feeder-free conditions. Reviewers did not consider this a major problem since the lines are not intended for clinical use. - The proposed hiPSC characterization methods are appropriate for the high throughput required for this project and will ensure the production of high quality lines. - Thaw recovery should be added to the hiPSC characterization tests. - Reviewers appreciated the proposed tests for mycoplasma contamination during the initial quarantine phase of the source cells. In addition, reviewers suggested that testing for microbial contamination be added, and that the applicant consider mycoplasma and microbial hiPSC release testing, especially if this is not performed on feeder cells. - The applicant does not discuss failure rates. Documentation and Quality Control - The quality control (QC) and documentation processes proposed are a strength of the proposal and documents and protocols are in place for most of the procedures that would be performed for this program. - The applicant institution is experienced in working under Standard Operating Procedures. Feasibility and Resources - The proposal to derive hiPSC lines from 1517 tissue donors does not meet the intended scope for RFA 12-03 (3000 donors). The applicant states that economies of scale may allow for a greater number of donors, but they cannot commit to that now. Reviewers considered whether the lower number of hiPSC lines translates into higher quality lines, as more funds would be spent per hiPSC line than in a proposal that includes 3000 tissue donors. Reviewers felt that at least some of the proposal’s limitations lie in the constraints imposed by the relatively low throughput of the proposed hiPSC clone selection process. - Although the applicant provided some evidence of feasibility in terms of hiPSC lines that have been generated and contracts that have been awarded, reviewers did express some concern about the lack of documented ability to scale up the existing process. - The proposed laboratory space is appropriate. - Reviewers appreciated that in addition to the equipment requested, a significant amount of already existing resources will be employed for this project. Project Director (PD) and Team - The track record of the PD is very strong and includes work with stem cells. The PD has been involved in biotechnology start-ups with a focus on biologic therapeutics for over ten years, and has been with the applicant institution for several years. - The commitment of the PD is adequate and the existing team should be able to hire staff and provide training and guidance to the new team members. Reasonable collaborations with other California-based academic institutions and companies are in place. - In addition to the PD, there are several experienced managers and scientists named on the proposal who will devote 100% of their time to this project. The team appears capable of doing the work as proposed; it consists of individuals who have worked together for some time and are familiar with the methods required to undertake the proposed work. - Several eminent stem cell researchers will support the applicant with their advice. - Although the applicant institution has several years of relevant experience, its focus is rather distinct from the proposed effort, raising some concern regarding the overall commitment of the company to this project. Budget - The overall budget for this project is realistic. The decision to generate hiPSCs from only 1517 donors, rather than 3000 tissue donors, is based on budgetary constraints. It is likely that some of the QC testing could be reduced thus allowing more hiPSC lines to be generated. - Reviewers appreciated that this applicant did not request indirect costs.