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Engineered tissue arrays to streamline deimmunized DMD gene therapy vectors

工程组织阵列可简化去免疫 DMD 基因治疗载体

基本信息

批准号：
10724882
负责人：
Guy Leary Odom
金额：
$ 41.11万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10724882
关键词：
3-Dimensional Actins Activities of Daily Living Address Affinity Aftercare Age Months Amino Acid Sequence Amino Acids Antigen Presentation Antigen-Presenting Cells Binding Biological Assay Body Weight Cardiac Cell Separation Child Clinical Clinical Trials Collection Complex Computer software Cytotoxic T-Lymphocytes Data Databases Disease Progression Dose Duchenne muscular dystrophy Dystrophin Echocardiography Engineering Epitopes Evaluation Excision Exclusion Exclusion Criteria Exons Fiber Future Gene Transduction Agent Gene therapy trial Generations Genetic Diseases Glycoproteins Hand Strength Haplotypes Head Helper-Inducer T-Lymphocyte Hindlimb Human Immune Immune response Immunity Immunologics Individual Magnetism Major Histocompatibility Complex Measures Modeling Muscle Muscle Fibers Muscular Atrophy Mutation Outcome Patients Peptides Performance Phase Phase I Clinical Trials Phenotype Progressive Disease Proteins Proteomics Protocols documentation Rattus Recombinant adeno-associated virus (rAAV)Relaxation Residual state Resolution Respiratory Diaphragm Role Running Selection Criteria Serious Adverse Event Signal Transduction Spectrin Striated Muscles Structure System T cell response T-Cell Activation T-Cell Receptor T-Lymphocyte T-Lymphocyte Epitopes Testing Therapeutic Effect Time Tissue Engineering Tissue Microarray Update Validation Variant Viral Vector adeno-associated viral vector cell mediated immune response clinically relevant data integration de-immunization delivery vehicle design disease phenotype early phase clinical trial gene therapy gene therapy clinical trial genomic data immunogenic immunogenicity in vivo induced pluripotent stem cell innovation intravenous administration mdx mouse mechanical properties micro-dystrophin mini-dystrophin miniaturize muscle engineering muscular system novel pre-clinical predicting response predictive modeling predictive tools prevent screening skeletal symposium three dimensional cell culture treadmill vector

项目摘要

A major driver for immunogenicity is the presence of T-cell epitopes within a protein sequence that can activate helper T cells, resulting in neutralization of the therapeutic effect. For DMD gene therapy, the possibility of T-cell responses to pre-existing (from revertant fibers) &/or post-treatment dystrophin epitopes has recently emerged as a serious treatment issue. In the past year 5 DMD patients treated with AAV-microdystrophin (µDys) have developed T cell mediated immune responses against dystrophin. Based on immunological data, in all cases the antigenic region was highly suggested to be encoded by exons 8-11.1 Additionally, to being observed in patients deleted for this region (updates provided at MDA and ASGCT 2022 scientific conferences). A key component of our approach takes advantage of the fact that a single amino acid can anchor a novel peptide for a preferred binding state for antigen presenting cells’ via its major histocompatibility complex (MHC) and subsequent T cell receptor (TCR). It is the TCR recognition which leads to downstream signaling cascades and immune sequelae. Alternatively, conservatively modified residue(s), can disfavor antigen presentation, preventing dystrophin- specific immunity. Thus, avoidance of a specific immune response can be achieved if key epitope anchor residue(s) necessary for antigen presentation are modified to disrupt MHC recognition. Here, our proposal addresses the fundamental role of immunogenicity against dystrophin that has been found in DMD patients. This proposal integrates deimmunization protocols and structure-based protein redesign via Rosetta-software, with streamlined functional testing via our magnetic tissue array (Mantarray) platform-allowing for daily 3D DMD contractile performance assessment in-a-dish. In turn, dystrophin epitopes can be predicted & ranked according to their antigenicity by correlating their amino acid sequence content with immunological databases. Known and predicted epitopes will be targeted for redesign to reduce or eliminate putative MHC-binding peptides and subsequent T cell activation. Our deimmunization approach will build on previous efforts while focusing on exons 8-11 of dystrophin, & integrating data as it becomes available through any DMD clinical trial alliance that unfolds. The exon 8-11 region, now part of Pfizers’ updated patient exclusion criteria, encodes a portion of the actin- binding domain, extending through spectrin-like repeat 1 (note: we have already deimmunized much of exons 6- 8). Importantly, restricting deimmunization efforts to exons 8-11 of dystrophin would be beneficial for all µDys designs currently in clinical trials. In summary, we propose to deliver deimmunized µDys vectors to human DMD 3D engineered muscle tissues and measure contractile force(s) via our Mantarray system in an effort to screen for highly functional leading candidates with reduced immunogenicity. To fully validate our 3D muscle system as a predictive model we will test optimized candidates(s) via rAAV vectors driven by highly muscle specific regulatory cassettes for their capacity to ameliorate disease progression in the DMDmdx rat model of DMD. These results will provide necessary preclinical outcome metrics required to advance towards Phase 1 clinical trials.

免疫原性的主要驱动因素是可以激活的蛋白质序列中t细胞表位的存在