Funding opportunities

Derivation of New iPS Cell Lines and Development of a Novel Stem Cell Platform for Parkinson's Research and Therapeutic Applications

Funding Type: 
New Cell Lines
Grant Number: 
RL1-00653
Funds requested: 
$1 266 332
Funding Recommendations: 
Not recommended
Grant approved: 
No
Public Abstract: 
Stem cells are unique in their ability transform into many different cell types. This has great potential value in the discovery of new drugs and therapies for many diseases. The creation of stem cells types (or lines) with specific genetic makeup that predisposes, or leads to disease, is an essential tool for the development of therapies for heritable diseases. It is also important to study stem cells representing a diversity of genetic backgrounds, since different patients respond differently to a given therapy. Unfortunately, growing stem cells and transforming them into different cell types is a slow and sensitive procedure, which slows the pace of stem cell research. Better technology is needed to carefully control the environment in which the cells grow, and to continuously monitor the changes in cells that occur during growth, or when exposed to various chemicals or drugs. To address these needs{REDACTED}, in collaboration with the Center for Regenerative Medicine at the {REDACTED}and {REDACTED}, proposes to develop a new research tool and apply it to create new stem cell lines for Parkinson’s research. In keeping with the intent of the CIRM program, the goals of this project are to: a) Create Parkinson’s disease-specific stem cell lines b) Develop a standard research platform for performing stem cell research c) Demonstrate the utility of the approach for Parkinson’s Research d) Disseminate the technology to the general stem cell research community to accelerate the pace of therapeutic breakthroughs for various diseases With the largest population in the US, California has millions of citizens that can potentially benefit from advances in stem cell research.
Statement of Benefit to California: 
{REDACTED} is developing a complete stem cell research system to serve as the standard platform to facilitate and accelerate the pace of stem cell-based drug discovery and therapeutic breakthroughs. In particular, the proposed project will apply this technology to Parkinson’s research. With the largest population in the US, California has millions of citizens that can potentially benefit from advances in stem cell research. As a California-based biotechnology company, {REDACTED} has regional access to, and close relationships with, the institutes that form the core of the nation’s stem cell research community. Thus, CIRM funds will be efficiently leveraged as {REDACTED} will be able to support and interact closely with collaborators (specifically the Center for Regenerative Medicine at {REDACTED}. This interaction will enable these researchers to more quickly adopt this cutting-edge technology to achieve their research goals faster, and more cost-effectively. In addition to the potential public health benefits for the citizens of California, this funding could generate tens of millions in tax revenues for the state. Experts estimate that the total market for stem cell research products will reach $10B over the next ten years. CIRM funding to enhance the {REDACTED} system and demonstrate its utility on the proposed Parkinson’s application will enable {REDACTED} to establish {REDACTED}as the standard platform for stem cell research. As a California-based company, {REDACTED}’s successful commercialization of the {REDACTED} system will translate directly into increased tax revenues for the state.
Review Summary: 
Executive Summary This proposal aims to create induced pluripotent stem (iPS) cell lines from normal and Parkinson’s Disease (PD) patients and then exploit the applicant’s new cell culture technology for improving culture of cells, their differentiation, and then disseminate the cells to the community for study. The principal investigator (PI) proposes to use both plasmid and viral vector methods for transfer of genes encoding reprogramming factors into fibroblasts. The project will be conducted in collaboration with two California institutions with expertise in stem cells and PD, respectively. All in all, the proposal focuses on improving reprogramming efficiency and cell culture/differentiation, an important goal to advance stem cell research. The new PD-specific iPS cell lines (12-15 planned) would be useful for disease-specific studies. In terms of feasibility, the strength of this proposal rests on the new technology, which is put forward as a greatly improved method for cell culture and cell differentiation. However, reviewers found that the applicant did not provide enough evidence and did not adequately describe how the new technology will increase iPS cell derivation efficiency or how it will be advantageous for studying neuronal differentiation when compared to standard methods. Other issues that call the feasibility of this proposal into question include the fact that the proposed characterization of iPS cells is limited to PCR-based gene expression analyses, and it does not appear as if the applicant plans to use any other type of evaluation for pluripotency. Moreover, preliminary data presented by the applicant show that cell culture results obtained using the new technology seem as good as those obtained with standard tissue culture dishes, and s/he does not provide evidence that the new technology is superior. Without this demonstration, it is difficult to see how this proposal will advance the field. Finally, the proposal does not address how direct reprogramming would advance the study of PD. Instead the proposal focuses on development and improvements of the applicant’s new technology. Reviewer One Comments Significance: Making iPS lines from PD patients could accelerate work in the field (assuming phenotypes can be established from differentiated cells). Improving methods for iPS generation are important as well. Feasibility: The strength of this proposal rests on the SmartSlide system, which is put forward as a greatly improved method for cell culture and cell differentiation. Unfortunately, there is precious little provided in the application to validate the system. Indeed, Figure 4, where Oct 4 expression is shown for mouse embryonic stem cells (mESCs), there is no difference between the SmartSlide and a standard 6-well plate. The SmartSlide approach seems as good as standard tissue culture dishes, but we are not shown evidence that it is superior. Without this demonstration, it is difficult to see how this proposal will advance the field. Responsiveness to RFA: Responsive. Reviewer Two Comments Significance: The investigators at WaferGen aim to demonstrate the utility of the SmartSlide System by deriving new Parkinson’s disease-specific iPS lines. PD iPS lines would be very useful for research use. Improving reprogramming efficiency and cell culture / differentiation would also be an important advance. Feasibility: The experience of the investigators and the facilities available are more than adequate. The proposal does not address how direct reprogramming would advance the study of Parkinson’s Disease. Instead the proposal focuses on how the SmartSlide system cell culture platform should improve direct reprogramming efficiency and the ability to culture and differentiate stem cells. The experiments proposed do not adequately describe how the SmartSlide system will increase iPS efficiency over standard methods or how it will be advantageous over standard methods for studying neuronal differentiation. Moreover, the preliminary results fail to demonstrate an advantage of SmartSlide system over standard mES cell culture. Responsiveness to RFA: The investigators will generate pluripotent stem cell lines. The lines they create will be shared and the SmartSlide system will be commercialized for use in stem cell line derivation / maintenance / differentiation. Reviewer Three Comments Significance: The proposal calls for the derivation of new iPS lines from normal and Parkinson patients, and development of a novel platform (WaferGen microincubation and imaging system) for research and therapeutic applications. The new iPS will be useful for disease specific studies (e.g. drug testing). Feasibility: Design and Feasibility They plan to create 12-15 disease-specific cell lines. They will start with facial dermal fibroblasts, using relatively standard culture conditions, and experiment with both viral and non-viral approaches (however, the methods only describe Lenti approach – 4 separate vectors). They will use their proprietary SmartSlide system to control culture conditions post-transduction (claim up to 80% efficiency, but minimal direct evidence provided). They will characterize iPS by PCR gene expression, there did not seem to be any other evaluation for pluripotency? They will differentiate iPS cells into neuronal cell types, and evaluate for oxidative phosphorylation defects. Most of the proposal describes optimizing the SmartSlide system (oxygen monitoring, pH control, multiplex micro-incubator format) which they plan to sell to the stem cell community as a tool. The first upgrade will be O2 monitor, followed by micro-well arrays – increase to 12 plex, then 24 plex ,and include pH control. They are planning to employ these upgraded SmartSlide to differentiate iPS cells into specific neuronal cells, then challenge with chemical agents and monitor responses such as survival and morphology. They would provide these iPS to the CIRM community for Parkinson studies. Strengths: Consultants have significant relevant experience Concerns: • There is a presumption that iPS from Parkinson’s will be different from normal, however no data was presented to support that hypothesis. • PI has no direct experience with iPS Responsiveness to RFA: Focus is clearly on development and improvements in the WaferGen SmartSlide system, not really on the derivation of new iPS (only Aim 1 of the proposal has the derivation of new iPS), and even this includes use of SmartSlide. The other three aims are totally SmartSlide biased.
Conflicts: 

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