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.

项目摘要尽管镰状细胞疾病（SCD）的医疗护理进展，大多数患者仍在经历使疼痛衰弱，生活质量差，进行性器官降低和过早死亡。我们正在开发一个基于自体造血干细胞基因组编辑的新型SCD治疗疗法（HSC）诱导红细胞（RBC）中胎儿血红蛋白（HBF，22）的产生。自然遗传变体可以产生高水平的HBF，以减轻或消除SCD的病理生理。我们出版了研究表明，这些变体之一可以通过Cas9介导的-珠蛋白基因的破坏来重新创建 Bcl11a识别的启动子图案，Bcl11a是一种转录复制蛋白，驱动正常围产期开关从珠蛋白到珠蛋白的表达。新的初步数据显示，正常或SCD患者捐助者的翻译用核糖核蛋白（RNP）复合物的CD34+细胞由Cas9和引导RNA（GRNA）组成 Bcl11a结合基序，然后进行异种移植到免疫缺陷小鼠中，始终达到≥70％在骨髓填充的HSC中进行靶向编辑，没有严格的基因组检测到的脱靶突变灵敏度为0.1％的广泛活性分析。修改后的HSC在体内生成了rbcs，> 30％的punchlular HBF和2％O2的HBF和18-31％的病态，而未修改的对照RBC中的HBF <5％和62-71％（p <0.0001）。现在，我们将通过设计和执行第一台 - 在人类临床研究，称为圣裘德自体基因组编辑的干细胞（SAGES1），检查了安全性根据3个目标，自体-珠蛋白启动子编辑的CD34+细胞（药物SJ-1001）的有效性。 AIM 1将定义SJ-1001的作用，效力和安全性机理，并通过FDA实现和探索性研究。 AIM 2将通过优化建立SJ-1001的CGMP临床规模制造过程开发，为临床试验生成药品释放分析，并将其转移圣裘德当前良好制造实践（CGMP）设施的协议。 AIM 3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。这将包括开发增强的多学科知情同意过程，安全性和效率评估以及治疗后的机械研究。我们的工作有可能挽救苦难并延长数千名严重SCD患者。