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（由申请人提供）：世界梦想用干细胞治愈每一种疾病。我们都知道这是不可能的。但是利用干细胞再生组织的主要障碍 与接受者的细胞在基因上不相同的干细胞容易产生免疫排斥反应。一种克服这一障碍的创新解决方案是使用患者自身的体细胞来产生干细胞，称为iPSC（诱导多能干细胞）。一旦产生，这些iPSC可以被给予患者而不用担心排斥，因为iPSC与患者体内的所有其他细胞在遗传上是相同的。然而，给予iPSC并不是大多数遗传性疾病的解决方案，因为产生的iPSC也含有致病的遗传缺陷。我们组建了一个拥有干细胞生物学、神经治疗学、转化科学和神经药理学专业知识的团队来解决这个问题。我们建议使用一种名为CRISPR-Cas9（clustered regularly interspaced short palindromic repeats-CRISPR associated protein 9）的新技术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来纠正从患者中分离的iPSC中的遗传缺陷。修正后的干细胞将被移植回患者体内，以提供具有基因功能拷贝的细胞。我们建议使用神经干细胞治疗遗传性儿童神经系统疾病粘多糖样变性IIIB型（MPS IIIB）来测试这一概念。基因校正的自体干细胞疗法非常适合MPS IIIB等疾病，这些疾病是由于缺乏分泌蛋白（在这种情况下，NAGLU [α-N-乙酰氨基葡萄糖苷酶]），因为非常少量的校正干细胞可以具有很大的治疗效果。我们还将采用糖基化非依赖性溶酶体靶向，其使用来自IGF 2的肽，其改善分泌的NAGLU进入大脑的摄取。在本提案的R21阶段，我们将进行体外研究，以确定我们校正的干细胞如何有效地分泌可被MPS IIIB细胞摄取的正常NAGLU和NAGLU-IGF 2。我们还将进行体内研究，以确定神经干细胞在新生动物和成年动物的三种不同剂量下在整个大脑中的分布情况。在提案的R33阶段，我们将把这些概念放在一起，用于校正神经干细胞对MPS IIIB疾病影响的体内研究。我们将评估生化、神经病理和神经行为结果。我们的目标是为这种毁灭性的、无法治愈的疾病创造一种永久性的治疗方法，并为治疗其他因缺乏功能性分泌蛋白而引起的遗传疾病建立一个平台。