The objective of this proposal is to develop an efficient and robust method to epigenetically reprogram naïve human T cells into regulatory T cells (Tregs) that can prevent graft versus host disease (GvHD). Our short term goal for Phase I of the STTR award is to demonstrate that we can epigenetically-reprogram naïve T cells using CRISPR/Cas9-based technology into immunosuppressive cells that have the protein marker, gene expression, and epigenetic regulatory profiles of Tregs. This will motivate Phase II of the STTR, where we will test the epigenetically-reprogrammed Tregs in a humanized mouse model of graft versus host disease. The data obtained during the STTR will position the therapy for clinical trials to prevent GvHD in hematopoietic cell transplantation. There is a clinical need to improve patient outcomes in allogeneic hematopoietic cell transplantation (HCT) due to the high incidence, morbiditity, and mortality associated with GvHD. Approximately half of all HCTs utilize hematopoietic cells from unrelated donors, and approximately 26,800 patients will undergo this procedure per year worldwide. Despite improved diagnostics to match the most immunogenic antigens on the donor cells with those of the recipient, up to 70% of allogeneic HCT recipients will develop GvHD as a result of their graft. Only 50% of HCT recipients that develop GvHD survive 3 years beyond the transplant procedure. GvHD is mediated by donor T cells that recognize polymorphisms in the protein-coding genome of the graft and the recipient, and these reactive T cells can be suppressed by Tregs. We will utilize CRISPR/Cas9-based epigenetic editing tools to reprogram naïve human T cells, which be easily collected from whole blood by apheresis or density gradient centrifugation, into immunosuppressive Tregs. We will first perform a high-throughput screens of candidate Treg- specific gene regulatory elements that we have identified through computational analysis of epigenomic data. We will then validate that these elements can be activated by CRISPR/Cas9-based epigenome modification tools that we have developed to generate T cells with the phenotype, in vitro suppressive properties, gene expression, and chromatin regulatory profile of Tregs. !
Public Health Relevance Statement: PROJECT NARRATIVE This STTR proposal seeks to identify gene regulatory elements that, when activated, allow T cells to adopt an immunosuppressive, regulatory function that is critical to prevent graft versus host disease (GvHD) after hematopoietic cell transplantation (HCT) from another individual. There is a significant clinical need for improved immunosuppressive therapies that can prevent GvHD because up to 70% of HCT patients develop GvHD and only 50% of those with the disease survive more than 3 years. In this STTR, we will reprogram graft T cells that are potentially reactive towards host antigens into regulatory T cells using tools that activate the expression of specific genes controlling the immunosuppressive function of these cells. !
Project Terms: Adopted; Adoptive Cell Transfers; Affect; Allogenic; Antigens; ATAC-seq; Award; base; Blood Component Removal; Cell physiology; Cell Therapy; Cell Transplants; Cells; Chimeric Proteins; Chromatin; Chromatin Structure; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Complex; Computer Analysis; CRISPR/Cas technology; Cyclosporins; Data; Density Gradient Centrifugation; Deposition; Diagnostic; Disease; Elements; Engineering; Enhancers; EP300 gene; Epigenetic Process; epigenome; epigenomics; Failure; Flow Cytometry; FOXP3 gene; Funding; Gene Expression; Gene Expression Profile; Genes; Genetic Polymorphism; Genome; genome-wide; Genomics; Goals; Graft-vs-Host Disease; Guide RNA; Hematopoietic; hematopoietic cell transplantation; high risk; high throughput screening; Histone Acetylation; histone acetyltransferase; HLA Antigens; Human; Human Genome; humanized mouse; Immune; Immune Tolerance; immunogenic; Immunosuppressive Agents; improved; In Vitro; Incidence; Individual; Inflammation; Inflammatory; innovation; Mediating; Methods; Mission; Modeling; Modification; mortality; mouse model; Opportunistic Infections; Patient-Focused Outcomes; Patients; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Phase; Phenotype; Positioning Attribute; prevent; Procedures; programs; Property; protein biomarkers; Proteins; Regulator Genes; Regulatory Element; Regulatory T-Lymphocyte; Sirolimus; Small Business Technology Transfer Research; Structure; success; Symptoms; T-Lymphocyte; Tacrolimus; Technology; Testing; Therapeutic immunosuppression; Therapy Clinical Trials; Tissues; tool; transcription factor; transcriptome sequencing; Transplantation; Whole Blood
