Of the 44 applications submitted to the NIH CRM Pilot Award Program for FY12 funding, a total of 6 projects were ultimately funded. This number is smaller than in previous years. Funding recommendations were based on rankings by thirteen extramural reviewers, and the projects were selected because they had had the potential to translate into clinical discoveries and would help build a critical mass of investigators working on stem cell projects. The recommendations were supported by leadership and the NIH CRM advisory groups. The investigators with new projects funded in FY12 represent the following institutes: NIDDK, NEI, NINDS, NCI and NHGRI. As in previous years, the newly funded projects focus on iPSCs yet cover a broad range of topics. As detailed below, topics included constructing iPSC-derived RPE tissue for therapy, using lineage reprogramming of tumor-specific T cells, using patient-specific iPS cells for progress in cell therapy for FPD/AML, making neurons from iPSCs to study disease pathology and screen for drugs, and generating disease models. In a study focused on hepatoctyes, human iPSC lines will be generated from primary human fibroblasts, and differentiated into human hepatocytesm for examination of their functions. Hepatocytes will be applied not only to clinical therapy of end-stage liver disease secondary to viral hepatitis, but also to genetic and metabolic liver diseases, such as 1-antitrypsin deficiency, hemochromatosis, nonalcoholic steatohepatitis, alcohol-related cirrhosis, and possibly pediatric liver disease of inborn error of metabolism. Ultimately, they plan to examine whether they can generate iPSCs, correct any genetic defect, and differentiate them into hepatocytes from these patients for potential cell therapy. In a study focused on screening, screening applications will employ RPE fully differentiated from induced pluripotent stem cells (iPSCs). As previously validated in pilot experiments, compounds will be validated and fully differentiated RPE will be produced from iPSC. In this phase they will continue generating the screening assay and develop iPSC-derived RPE for future cell therapies. Building on earlier work showing that fetal human/iPSC-derived RPE cells generate an extracellular matrix (ECM) that helps maintain the RPE as a polarized sheet, they are testing the ability of biodegradable substrates to support the growth of RPE/ECM tissue which would lead to a safe and more effective autologous/heterologous cell-based therapy for patients. In a study focused on Fragile X syndrome (FX), fully reprogrammed iPSCs will be developed from FX patients using modified reprogramming strategies based on earlier gene silencing findings. FX iPSCs that show partial FMR1 reactivation can be differentiated into neurons, which will be useful for studying some aspects of disease pathology. They also intend to carry out a primary screen of an FDA-approved drug library to identify compounds capable of reversing FX gene silencing. This will be beneficial not only for understanding FMR1 gene silencing but will also be useful for understanding the factors necessary for complete reprogramming of human iPSCs. In a study focused on neurocognition, induced pluripotent stem cells (iPSCs) derived from patient fibroblasts will be differentiated into neurons. The study will characterize the neuronal cellular and synaptic phenotype, and undertake genetic and molecular manipulations to show that the synaptic phenotype is dependent on abnormal glycosylation of dystroglycan (DG) and can be modulated by interventions. Candidate compounds can then be screened for their translational potential to treat the neurocognitive impairment in patients with dystroglycanopathy. This project has additional significance as it would demonstrate that the synaptic dysfunction underlying a developmental form of intellectual disability and autistic spectrum disorder can be influenced by molecular treatment approaches. In a study focused on T cells, regenerative medicine techniques will be used that involve microRNA (miR) and transcription factor (TF)-based lineage reprogramming of T cells into younger forms. The study will focus reprogramming candidates identified in a FY10 project, which are differentially expressed in stem cell-like T cells and effector T cells. The study will now employ a zero-footprint adenoviral approach, evaluating results using highly multicolored flow cytometry, GFP reporter (in mice), and tumor treatment assays. This clinical trial could be among the first to employ regenerative medicine technology in a therapeutic setting in patients with cancer. And finally, in a study targeting Familial Platelet Disorder with propensity to acute myeloid leukemia (FPD/AML), iPSCS from family members will be derived, the mutant RUNX1 allele corrected using zing fingers, and iPSCS differentiated into HSCs for transplantation. If successful, the HSCs could reconstitute the hematopoietic system. In addition, to increase the efficiency iPSC differentiation to HSCs, a hematopoietic iPSC reporter line will be generated for small molecule and transcription factor screens to identify candidate molecules and genes directing hematopoietic differentiation. Of note, an EIR has already resulted from this last project. Through these continuing efforts, NIH CRM has been able to distribute seed funding appropriately for stem cell-based projects with tangible results, within a forum for sharing their work and participating in fruitful discussion.
