Targeted Enzyme Replacement Therapy for Rare Forms of Osteogenesis Imperfecta

靶向酶替代疗法治疗罕见的成骨不全症

基本信息

批准号：
8710973
负责人：
CAROLE L. CRAMER
金额：
$ 22.34万
依托单位：
BIOSTRATEGIES, LC
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8710973
关键词：
2-oxoglutarate 3-dioxygenase proline Address Adult Affect Affinity Binding Biodistribution Bone Density Bone Diseases Bone Resorption Carrier Proteins Cartilage Cell Lineage Cell surface Cells Chimeric Proteins Chondrocytes Codon Nucleotides Collagen Collagen Type I Complex Connective Tissue Connective Tissue Cells Connective Tissue Diseases Defect Disease Documentation Drug Delivery Systems Ear Endocytosis Enzyme Stability Enzymes Evaluation Exhibits Eye Fabry Disease Feasibility Studies Fibroblasts Gaucher Disease Genes Genetic Glycolipids Glycoproteins Goals Health Hereditary Disease Human Hydroxylation In Vitro Lead Lectin Legal patent Mammalian Cell Mediating Mesenchymal Mesenchyme Modeling Molecular Chaperones Molecular Genetics Monitor Mus Neuraxis Osteoblasts Osteogenesis Imperfecta Outcome Pathology Patients Pharmaceutical Preparations Pharmacologic Substance Phase Phenotype Plague Plant Lectins Plants Process Procollagen Production Proteins Replacement Therapy Retrieval Ricin Rough endoplasmic reticulum Safety Scaffolding Protein Signal Transduction Skeletal system Skin Small Business Innovation Research Grant Structure System Technology Testing Therapeutic Tissues Toxicology Variant base bisphosphonate bone bone mass cell type cost disease phenotype enzyme activity enzyme replacement therapy glucosylceramidase in vivo innovation malformation meetings mouse model novel novel therapeutics osteogenic outcome forecast patient population phase 2 study plant growth/development pre-clinical preference protein complex protein function research and development restoration skeletal therapeutic target trafficking transcytosis uptake

项目摘要

DESCRIPTION (provided by applicant): The long-term goal of this project is to develop an effective protein replacement therapeutic (PRT) for patients affected with rare forms of Osteogenesis imperfecta, OI-VIII (deficiency in prolyl 3-hydroxylase 1; P3H1) and OI-VII (deficiency in cartilage associated protein CRTAP). These PRTs will integrate the safety and cost advantages of plant-based bioproduction with innovations that enhance delivery and disease correction. OI is a group of genetic connective tissue disorders characterized by low bone mass and increased bone fragility. Current treatment options for OI are limited to bisphosphonates treatments, which inhibit bone resorption but are controversial due to the likelihood of deleterious effects in young patients as well as adults. Developing effective PRTs for these patients is challenging since classic enzyme replacement therapies (ERTs) have been relatively ineffective in correcting bone-related pathologies. PRTs for these rare OI diseases will require efficient delivery to multiple mesenchyme-derived cell lineages as well as effective targeting to rough endoplasmic reticulum (rER) to correct collagen malformation. To expand the delivery of corrective protein to critical cells and tissues, BioStrategies LC has been testing the potential of several plant lectins to function as effective carriers of PRT's into disease cells. These lectins have high affinity for glycoproteins and glycolipids common on mammalian cell surfaces and mediate efficient cellular uptake, transcytosis, and delivery to target tissues. We have demonstrated that one of these lectins, RTB, efficiently carries large fused model proteins into multiple cells of the skeletal systems including chondrocytes, osteoblasts, and mesenchymal cells. We also showed that RTB, modified to contain a KDEL ER-retrieval signal (RTBER), effectively directs payload proteins to the ER following endocytosis. We hypothesize that our fusion proteins will access cells of the skeletal system and will deliver PRTs to correct collagen defects in OI-VIII and OI-VII. The goal of this SBIR Phase I feasibility study is to test the potential of plants to produce RTBER fusions with P3H1 and/or CRTAP that are taken up into osteogenic cells, mobilized to the rER, and function to correct collagen processing defects at the cellular level. Specific aims for Phase I are 1) to produce and characterize RTB:collagen-modifying protein fusions using a transient plant-based expression platform, and 2) to demonstrate product efficacy in vitro by disease phenotype correction in OI cells. Based on successfully meeting the Phase I milestones of in vitro cell correction in either OI-VIII or OI- VI models of rare recessive OI, follow-on Phase II studies would focus on in vivo efficacy in mouse models of these OI diseases. These studies would include short-term biodistribution analyses and skeletal phenotype evaluation following extended replacement therapy treatments in either P3H1 deficient or Crtap-/- mice leading to Phase IIB toxicology assessment and other preclinical endpoints in support of an IND application at FDA.

描述（由申请人提供）：该项目的长期目标是为患有罕见形式的成骨不全症 OI-VIII（脯氨酰 3-羟化酶 1 缺乏；P3H1）的患者开发一种有效的蛋白质替代疗法 (PRT)。和 OI-VII（软骨相关蛋白 CRTAP 缺乏）。这些 PRT 将把基于植物的生物生产的安全性和成本优势与增强交付和疾病纠正的创新相结合。成骨不全症是一组遗传性结缔组织疾病，其特征是骨量低和骨脆性增加。目前成骨不全的治疗选择仅限于双磷酸盐治疗，这种治疗会抑制骨吸收，但由于可能对年轻患者和成人产生有害影响而存在争议。为这些患者开发有效的 PRT 具有挑战性，因为经典的酶替代疗法 (ERT) 在纠正骨相关病理方面相对无效。针对这些罕见 OI 疾病的 PRT 将需要有效递送至多种间充质衍生细胞谱系以及有效靶向粗面内质网（rER）以纠正胶原蛋白畸形。为了扩大纠正蛋白向关键细胞和组织的输送，BioStrategies LC 一直在测试几种植物凝集素作为 PRT 进入疾病细胞的有效载体的潜力。这些凝集素对哺乳动物细胞表面常见的糖蛋白和糖脂具有高亲和力，并介导有效的细胞摄取、转胞吞作用和递送至靶组织。我们已经证明，其中一种凝集素 RTB 可以有效地将大型融合模型蛋白携带到骨骼系统的多个细胞中，包括软骨细胞、成骨细胞和间充质细胞。我们还表明，经过修饰以包含 KDEL ER 检索信号 (RTBER) 的 RTB，可以在胞吞作用后有效地将有效负载蛋白引导至 ER。我们假设我们的融合蛋白将进入骨骼系统的细胞，并提供 PRT 以纠正 OI-VIII 和 OI-VII 中的胶原蛋白缺陷。这项 SBIR 第一阶段可行性研究的目标是测试植物与 P3H1 和/或 CRTAP 产生 RTBER 融合的潜力，这些融合被吸收到成骨细胞中，动员到 rER，并在细胞水平上纠正胶原蛋白加工缺陷。第一阶段的具体目标是 1) 使用基于植物的瞬时表达平台生产和表征 RTB：胶原蛋白修饰蛋白融合物，2) 通过 OI 细胞中的疾病表型校正来证明产品的体外功效。基于在罕见隐性 OI 的 OI-VIII 或 OI-VI 模型中成功实现体外细胞校正的 I 期里程碑，后续的 II 期研究将重点关注这些 OI 疾病小鼠模型的体内疗效。这些研究将包括在 P3H1 缺陷或 Crtap-/- 小鼠中进行长期替代治疗后的短期生物分布分析和骨骼表型评估，从而导致 IIB 期毒理学评估和其他临床前终点，以支持 FDA 的 IND 申请。