Novel therapeutic gene editing to induce fetal hemoglobin for sickle cell disease

诱导胎儿血红蛋白治疗镰状细胞病的新型治疗性基因编辑

基本信息

批准号：
10587901
负责人：
Akshay Sharma
金额：
$ 118.3万
依托单位：
ST. JUDE CHILDREN'S RESEARCH HOSPITAL
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 2028-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10587901
关键词：
AMD3100 Acute Pain Adult Affect Allogenic American Attention Autologous BCL11A gene Binding Bioethics Consultants Biological Assay Biological Markers Birth Blood Bone Marrow CD34 gene CRISPR/Cas technology Caring Cell Differentiation process Cells Cessation of life Chromosomal Rearrangement Clinical Clinical Research Clinical Trials Communities Complex Data Deterioration Development Disease Education Engineering Engraftment Enhancers Erythrocytes Erythroid Erythropoiesis Euthanasia Exhibits Fetal Hemoglobin Functional disorder Future Genes Genetic Genetic Diseases Genetic Transcription Globin Good Manufacturing Process Guide RNA Hematopoietic stem cells Hemoglobin Human Immune Immunodeficient Mouse Impairment In Vitro Individual Informed Consent Infusion procedures Laboratory Scientists Lead Life Longevity Mediating Medical Missense Mutation Modification Molecular Monitor Multiple Organ Failure Mus Mutation Detection Normal Cell Nuclear Localization Signal Organ Outcome Pain Parents Participant Patients Perinatal Pharmaceutical Preparations Pharmacology Study Polymers Premature Mortality Process Process Assessment Production Protocols documentation Publishing Quality Control Quality of life Repressor Proteins Research Ribonucleoproteins Running Safety Saint Jude Children&apos s Research Hospital Sickle Cell Anemia Sickle Cell Trait Sickle Hemoglobin Specific qualifier value Technology Testing Therapeutic Toxic effect Toxicology Transcription Repressor Transfection Translating Transplantation Variant Work Xenograft procedure analytical method bench to bedside beta Globin biophysical properties chronic pain clinical practice curative treatments debilitating pain design early phase clinical trial efficacy evaluation erythroid differentiation experience feasibility testing first-in-human gene therapy genetic variant genome editing genome-wide genotoxicity hematopoietic cell transplantation improved in vivo indexing insertion/deletion mutation manufacture manufacturing process multidisciplinary novel novel therapeutics off-target mutation patient oriented polymerization pre-clinical premature process optimization promoter research study safety assessment scale up sickling stem cell growth stem cell therapy stem cells therapeutic gene

项目摘要

PROJECT SUMMARY Despite advances in the medical care of sickle cell disease (SCD), most patients continue to experience debilitating pain, poor quality of life, progressive organ deterioration, and premature death. We are developing a novel, potentially curative therapy for SCD based on genome editing of autologous hematopoietic stem cells (HSCs) to induce the production of fetal hemoglobin (HbF, 22) in red blood cells (RBCs). Natural genetic variants can generate high levels of HbF that alleviate or eliminate the pathophysiology of SCD. Our published studies show that one of these variants can be recreated by Cas9-mediated disruption of a -globin gene promoter motif recognized by BCL11A, a transcriptional repressor protein that drives the normal perinatal switch from -globin to -globin expression. New preliminary data show that transfection of normal or SCD patient donor CD34+ cells with ribonucleoprotein (RNP) complex consisting of Cas9 and guide RNA (gRNA) targeting the BCL11A binding motif, followed by xenotransplantation into immunodeficient mice, consistently achieved ≥70% on-target editing in bone marrow-repopulating HSCs, with no off-target mutations detected by rigorous genome- wide activity analysis at a sensitivity of 0.1%. The modified HSCs generated RBCs in vivo with >30% pancellular HbF and 18-31% sickling in 2% O2, compared to <5% HbF and 62-71% sickling in unmodified control RBCs (P<0.0001). We will now translate our findings “from bench to bedside” by designing and carrying out a first-in- human clinical study, termed St. Jude Autologous Genome Edited Stem Cells (SAGES1), examining the safety and efficacy of autologous -globin promoter-edited CD34+ cells (drug product SJ-1001) according to 3 Aims. Aim 1 will define the mechanism of action, potency, and safety profile of SJ-1001 through FDA-enabling and exploratory research studies. Aim 2 will establish cGMP clinical scale manufacturing of SJ-1001 by optimizing process development, generating drug product release assays for the clinical trial, and transferring these protocols to the St. Jude current good manufacturing practice (cGMP) facility. Aim 3 will establish and manage the SAGES1 clinical trial to evaluate one-time SJ-1001 infusion as a cure for SCD. This will include the development of an enhanced multidisciplinary informed consent process, safety and efficacy assessments, and post therapy mechanistic studies. Our work has the potential to relieve suffering and extend the lifespan of thousands of patients with severe SCD.

项目概要尽管镰状细胞病（SCD）的医疗护理取得了进步，但大多数患者仍然经历使人衰弱的疼痛、生活质量差、进行性器官恶化和过早死亡。基于自体造血干细胞基因组编辑的 SCD 新型潜在治愈疗法 (HSC) 诱导红细胞 (RBC) 中胎儿血红蛋白 (HbF, 22) 的产生天然遗传。变异体可以产生高水平的 HbF，从而减轻或消除我们发表的 SCD 的病理生理学。研究表明，其中一种变体可以通过 Cas9 介导的 γ-珠蛋白基因破坏来重建被 BCL11A 识别的启动子基序，BCL11A 是一种驱动正常围产期开关的转录抑制蛋白从 -珠蛋白到 -珠蛋白表达新的初步数据表明，正常或 SCD 患者供体的感染。 CD34+ 细胞具有核糖核蛋白 (RNP) 复合物，该复合物由 Cas9 和引导 RNA (gRNA) 组成，靶向 BCL11A 结合基序，然后异种移植到免疫缺陷小鼠中，始终达到 ≥70% 骨髓再生 HSC 中的定点编辑，严格的基因组检测未检测到脱靶突变修饰后的 HSC 能够以 0.1% 的灵敏度进行广泛的活性分析，在体内生成的红细胞具有 >30% 的全细胞成分。 2% O2 中的 HbF 和 18-31% 镰状细胞，相比之下，未经修饰的对照红细胞中 <5% HbF 和 62-71% 镰状细胞（P<0.0001）我们现在将通过设计和实施首创将我们的发现“从实验室转移到临床”。人类临床研究，称为圣裘德自体基因组编辑干细胞（SAGES1），检查安全性以及根据 3 个目标的自体 γ-珠蛋白启动子编辑的 CD34+ 细胞（药品 SJ-1001）的功效。目标 1 将通过 FDA 的授权和批准来定义 SJ-1001 的作用机制、效力和安全性。目标 2 将通过优化建立 SJ-1001 的 cGMP 临床规模生产。工艺开发，生成用于临床试验的药物产品释放分析，并将这些分析转移 St. Jude 现行良好生产规范 (cGMP) 设施的协议将建立和管理。 SAGES1 临床试验评估一次性 SJ-1001 输注作为 SCD 的治疗方法。制定强化的多学科知情同意流程、安全性和有效性评估，以及我们的工作有可能减轻患者的痛苦并延长患者的寿命。数以千计的严重 SCD 患者。