Basic Biology V
$1 327 500
Recent advances in creating iPS cells from patients have opened up new doors to understand the pathophysiology of diseases and to discover new therapeutics. To unravel new signaling mechanisms involved in stem cell fate, we propose to investigate kinase functions in iPSC and differentiation. In aim 1, the role of cytoskeletal remodeling in pluripotency and stem cell differentiation will be investigated. We will probe the function of kinases by controlling their expression and function using pharmacological and RNAi methods. These findings will reveal mechanisms that regulate cellular shape and migration to create specific microenvironments for iPSC. In aim 2, nodes in networks of kinome will be perturbed and their effect on iPSC generation will be analyzed. Functional studies will be performed to identify the key enzyme substrates of specific pathways in inducing pluripotency and how the enzymatic reactions influence the network. In aim 3, the role of signaling network in sporadic and familial Alzheimer’s disease will be studied using iPS cells. We will develop a chemical biology strategy to identify proteins that are phosphorylated in disease states and regulate signals in neurons. When obtained results are compared and integrated with control iPSC, new mechanisms will provide insight into disease pathogenesis and new targets to develop patient-specific drugs in alleviating or slowing the progression of disease as well as new diagnostics to monitor the stages of disease.
Statement of Benefit to California:
The proposed research is likely to benefit the state of California in the following ways. (1) Understanding the signaling mechanisms to create a pluripotent state from somatic cells will create new knowledge that can be applied to generate new iPSC lines for disease modeling and developing cell based therapies. (2) Developing mechanism-based iPS technology in creating phenotypes relevant to familial and sporadic Alzheimer’s disease, which could take decades to study in patients, would provide new research and development opportunities. (3) The new insight from Alzheimer disease models can be useful in understanding the disease and developing patient-specific drugs for other neurodegenerative diseases. (4) New biomarkers can be developed to monitor the onset or progression of disease states. (4) Future capabilities to make new neurons in test tube will benefit in curing neuronal loss in disease and injuries. (5) Future work force will be trained in state of the art technologies required for the state to stay competitive in new economies.
This Track 1 (Fundamental Mechanisms) proposal is focused on the role of kinases in pluripotency and differentiation. The PI suggests that the findings from these studies will also provide new information regarding molecular mechanisms for Alzheimer’s disease. There are three aims proposed:  examine the role of specific kinases in stem cell generation and in cytoskeletal remodeling and cellular migration during reprogramming;  determine, using a kinome interaction network, the kinase signaling network that modulates stem cell generation; and  utilize stem cells derived from Alzheimer’s patients to investigate the role of the identified kinase network during neuronal differentiation. Significance and Innovation - The significance of the proposed studies is not clear and no innovative technologies or strategies are proposed. At best, these studies might contribute to an enhanced efficiency of reprogramming somatic cells, but this is not a goal of the RFA. - The examination of the role of kinases in Alzheimer’s is interesting, but not novel. - The project is unlikely to reveal any new mechanistic insights regarding Alzheimer’s disease. Feasibility and Experimental Design - The experimental design for Aims 1 and 2 is clear, but attempts to accomplish more than is feasible in a 3-year timeline. - The PI does not indicate the number of time points to be used in the transition of pluripotency to neurogenesis. Given that it takes greater than 10 weeks to differentiate pluriopotent cells to the type of neurons that are the targets for the disease (with an unknown number of transition states), there may be an infinite number of time points to consider. - The preliminary data are generally substantive and supportive of the proposal, although preliminary data for Aim 3 is lacking. - Specific Aim 3 appears to have no connection to the first two aims and there is no specific strategy described for isolating “neurons” that are affected by Alzheimer’s disease. - Alternative approaches are not clearly indicated for the proposed studies. Principal Investigator (PI) and Research Team - The PI is an established investigator with expertise in RNA and chemical biology. - The collaborating investigators have a strong emphasis in neurobiology, cell biology and electron microscopy imaging. - The commitment by the co-investigators (1%) appears to be inadequate, given the central role that they will play in this project. Responsiveness to the RFA - The project is responsive to the RFA.