Modified neural stem cells for glycosylation-independent treatment of MPS IIIB

用于 MPS IIIB 糖基化独立治疗的修饰神经干细胞

• 批准号: 9089175
• 负责人: Michelina Iacovino
• 金额: $ 26.06万
• 依托单位: LUNDQUIST INSTITUTE FOR BIOMEDICAL INNOVATION AT HARBOR-UCLA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2018-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9089175
• 关键词: Adult; Age; Animal Model; Area; Autologous; Back; Behavior; Biochemical; Biochemistry; Brain; Cells; Cerebral Ventricles; Childhood; Clinic; Clustered Regularly Interspaced Short Palindromic Repeats; Data; Dementia; Derivation procedure; Development; Diffuse; Disease; Dose; Dreams; Fibroblasts; Fright; Genes; Goals; Hereditary Disease; IGF2 gene; Immune; Immune response; In Vitro; Inflammation; Inherited; Injection of therapeutic agent; Laboratories; LacZ Genes; Lateral; Liquid substance; Mucopolysaccharidoses; Mus; Mutation; N acetylglucosaminidase; Natural regeneration; Neonatal; Neuraxis; Neurodegenerative Disorders; Neurologic; Neuropharmacology; Newborn Animals; Normal Cell; Oncogenes; Other Genetics; Outcome; Pathology; Patients; Peptides; Phase; Problem Solving; Proteins; Research Institute; Somatic Cell; Source; Spinal Cord; Stem cells; Subfamily lentivirinae; System; Techniques; Technology; Testing; Therapeutic; Therapeutic Effect; Tissues; Transfection; Transgenes; Translational Research; Transplantation; Treatment Efficacy; Vent; brain research; cell preparation; effective therapy; enzyme deficiency; experience; gene correction; glycosylation; graft vs host disease; improved; in vivo; induced pluripotent stem cell; innovation; mature animal; nerve stem cell; nervous system disorder; neurobehavioral; new technology; pre-clinical; public health relevance; repaired; response; stem cell biology; stem cell differentiation; stem cell therapy; success; uptake; vector

 DESCRIPTION (provided by applicant): The world dreams of curing every disease with stem cells. We all know this is unlikely. But the main barrier to using stem cells to regenerate tissues and treat diseases is that stem cells that are not genetically identical to the recipient's cells ae prone to immune rejection. An innovative solution to overcome this barrier uses the patient's own somatic cells to generate stem cells called iPSCs (induced pluripotent stem cells). Once generated, these iPSCs can be administered to the patient without fear of rejection because iPSCs are genetically identical to all other cells in the patient. However, administering iPSCs is not a solution for most inherited diseases because the generated iPSCs also contain the disease-causing genetic defect. We have assembled a team with expertise in stem cell biology, neurotherapeutics, translational science, and neuropharmacology to solve this problem. We propose to use a new technology called CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9) to repair the faulty gene in the recipient's stem cells. CRISPR-Cas9 is the most efficient gene correction system to date. Our approach will use CRISPR-Cas9 to correct the genetic defect in iPSCs isolated from patients. The corrected stem cells will be transplanted back to the patient to provide cells that have a functioning copy of the gene. We propose to test this concept using neural stem cells to treat the inherited childhood neurological disease mucopolysaccharidosis type IIIB (MPS IIIB). Gene-corrected, autologous stem cell therapy is ideally suited to diseases like MPS IIIB, which are due to lack of a secreted protein (in this case, NAGLU [alpha-N-acetylglucosaminidase]), because a very small number of corrected stem cells can have a large therapeutic effect. We will also employ glycosylation-independent lysosomal targeting using a peptide derived from IGF2 that improves the uptake of secreted NAGLU into the brain. In the R21 phase of this proposal, we will perform in vitro studies to determine how effectively our corrected stem cells secrete normal NAGLU and NAGLU-IGF2 that can be taken up by MPS IIIB cells. We will also perform in vivo studies to determine how well the neural stem cells distribute throughout the brain at three different doses, in neonatal and adult animals. In the R33 phase of the proposal, we will put these concepts together for in vivo studies of the effect of corrected neural stem cell on MPS IIIB disease. We will assess biochemical, neuropathological, and neurobehavioral outcomes. The goal is to create a permanent treatment for this devastating, untreatable disease, and to build a platform for treating other genetic diseases that are caused by the lack of a functioning, secreted protein.

 描述（由适用提供）：世界梦想着用干细胞制造每种疾病。我们都知道这不太可能。但是使用干细胞再生组织的主要障碍 治疗疾病的是，与受体细胞不易免疫排斥的干细胞与受体细胞相同。克服该障碍的创新解决方案使用患者自己的体细胞产生称为IPSC（诱导多能干细胞）的干细胞。一旦生成，这些IPSC就可以对患者施用，而不必担心被排斥，因为IPSC在遗传上与患者中的所有其他细胞相同。但是，对大多数遗传疾病的施用并不是解决方案，因为产生的IPSC还包含引起疾病的遗传缺陷。我们已经组建了一个在干细胞生物学，神经疗法，转化科学和神经药理学方面具有专业知识的团队，以解决这个问题。我们建议使用一种称为CRISPR-CAS9的新技术（群集定期间隔短的短质体重复 - crispr相关蛋白9）来修复受体干细胞中的错误基因。 CRISPR-CAS9是迄今为止最有效的基因校正系统。我们的方法将使用CRISPR-CAS9纠正从患者中分离出的IPSC的遗传缺陷。校正后的干细胞将被移植回患者，以提供具有该基因功能副本的细胞。我们建议使用神经元干细胞来测试这一概念，以治疗IIIB型遗传性神经系统疾病粘二糖（MPS IIIB）。由基因校正的自体干细胞疗法非常适合MPS IIIB等疾病，由于缺乏分泌的蛋白质（在这种情况下，Naglu [alpha-n-乙酰葡萄糖酰胺酶）），因为少数校正的干细胞可能具有较大的治疗效果。我们还将使用源自IGF2的肽采用糖基化独立的溶酶体靶向，从而改善了分泌的Naglu对大脑的摄取。在该提案的R21阶段，我们将进行体外研究，以确定如何有效地校正的干细胞秘密正常Naglu和Naglu-igf2，这些naglu-igf2可以被MPS IIIB细胞占用。我们还将进行体内研究，以确定神经干细胞以三种不同剂量在新生儿和成年动物中分布在整个大脑中的分布程度。在提案的R33阶段，我们将将这些概念放在体内研究中，以研究校正神经元干细胞对MPS IIIB疾病的影响。我们将评估生化，神经病理学和神经性的结果。目的是为这种毁灭性，不可治疗的疾病创建永久治疗方法，并建立一个平台，以治疗由于缺乏功能正常的分泌蛋白质引起的其他遗传疾病。