Microfluidic cell squeezing platform for the transdifferentiation of somatic cells for efficient generation of a cell replacement therapy for Parkinsons Disease

用于体细胞转分化的微流控细胞挤压平台，可有效生成帕金森病的细胞替代疗法

• 批准号: 10483308
• 负责人: Devin Bridgen
• 金额: $ 100.14万
• 依托单位: SQZ BIOTECHNOLOGIES COMPANY, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-14 至 2022-12-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10483308
• 关键词: Affect; Age related macular degeneration; Allogenic; Animal Model; Autologous; Biopsy; Blood Cells; Cell Differentiation process; Cell Therapy; Cell Transplantation; Cells; Chronic; Clinic; Clinical; DNA Sequence Alteration; Data; Degenerative Disorder; Development; Disease; Disease model; Dose; Electrophysiology (science); Electroporation; Engineering; Engraftment; Ensure; Failure; Gene Expression; Generations; Genetic; Genetic Risk; Goals; Health; Heart failure; Human; Human Engineering; Immune; Immunosuppression; In Vitro; Insulin-Dependent Diabetes Mellitus; Legal patent; Liver Failure; Macular degeneration; Mammalian Cell; Messenger RNA; Methods; Microfluidics; Modeling; Modification; Molecular; Morphology; Mus; Nerve Degeneration; Neurons; Nucleic Acids; Organ; Oxidopamine; Parkinson Disease; Patient-Focused Outcomes; Patients; Peptides; Peripheral Blood Mononuclear Cell; Pharmacologic Substance; Phase; Physiological; Play; Pluripotent Stem Cells; Population; Pre-Clinical Model; Process; Production; Proteins; Quality of life; Rattus; Research; Risk; Safety; Signal Pathway; Small Business Innovation Research Grant; Solid Neoplasm; Somatic Cell; Symptoms; System; Technology; Therapeutic; Thromboplastin; Time; Tissues; Translating; Transplantation; Viral Vector; Work; age related; base; biopharmaceutical industry; cell injury; cell replacement therapy; cell transformation; cell type; clinical translation; clinically relevant; dopaminergic neuron; improved; in vivo; in vivo engraftment; induced pluripotent stem cell; lipofection; macula; neurotransmission; phase I trial; prevent; programs; reduce symptoms; stem cells; transcription factor; transdifferentiation; viral gene delivery; virtual delivery

PROJECT SUMMARY: Major diseases, such as heart failure, Parkinson’s Disease (PD), type 1 diabetes, and age-related macular de- generation, are examples of organ and systemic failure driven by damage to specific cell types. One possible therapeutic solution is to replace the damaged cells with ex vivo engineered, physiologically functional cells to alleviate clinical symptoms. However, expensive, time-intensive, and inefficient cell reprogramming methods for generating transplantable therapeutic cells for degenerative disorders hinders development and clinical transla- tion of potentially transformative therapies. Our goal is to develop a highly efficient process for producing cell replacement therapies that can be reliably manufactured at-scale to treat currently intractable diseases such as Parkinson’s Disease. This goal will build upon our patented and proven Cell Squeeze® technology that can deliver materials including mRNA, proteins, and peptides into sensitive primary cells. For this Phase II SBIR proposal, our overall objective is to demonstrate that with Cell Squeeze® technology we can introduce transcrip- tion factors that can increase the efficiency of transdifferentiating peripheral blood cells (PBMCs) into clinically relevant dopaminergic neurons. Our central hypothesis is that we can precisely control the timing, dose, and combinations of transcription factors to create high quality, functional cell products in greater quantities in a shorter time than is possible with current methods and free of risks associated with viral gene delivery. Support- ing this goal, we have already demonstrated that the squeeze treatment alone does not significantly affect gene expression, we can efficiently generate neurons from iPSCs, and we can introduce multiple transcription factors into PBMCs to upregulate expression of key neuronal signaling pathways. The rationale is that our non-viral method of delivering transcription factors to drive cell fate could significantly improve the efficiency and efficacy of cells produced with fewer safety and regulatory concerns as compared to other methods of transdifferentiation. Furthermore, in comparison to allogeneic iPSC derived products, autologous cells would not require chronic immunosuppression – a key factor to ensure long term health of the patient. In Aim 1, we seek to optimize methods for the Cell Squeeze® technology to deliver mRNA-based transcription factors to PBMCs to drive effi- cient transdifferentiation into dopaminergic neurons (DNs). Resultant DNs will be thoroughly characterized in vitro and compared to DN generated from iPSC using existing methods. In Aim 2, these DNs will be functionally assessed in an in vivo murine PD model to support the further development into a potentially transformative cell therapy. Successful completion of these aims may support partnering opportunities with other biopharmaceutical companies who are seeking differentiated cell therapy approaches in neurodegeneration.

项目摘要： 主要疾病，例如心力衰竭，帕金森氏病（PD），1型糖尿病以及与年龄有关的黄斑de- 产生是由特定细胞类型损害驱动的器官和系统故障的例子。一个可能 治疗解决方案是用离体工程的，物理功能的细胞代替受损的细胞 减轻临床症状。但是，昂贵，耗时且效率低下的单元重编程方法 产生可移植的治疗细胞用于退行性疾病，会阻碍发展和临床翻译 潜在的转化疗法的影响。我们的目标是开发一个高效生产细胞的过程 可以可靠的替代疗法，以治疗当前棘手的疾病，例如 帕金森氏病。这个目标将建立在我们获得专利的CellSqueeze®技术的基础上 将包括mRNA，蛋白质和胡椒粉在内的材料输送到敏感的原代细胞中。对于此II阶段SBIR 提案，我们的总体目标是证明，通过CellSqueeze®技术，我们可以引入笔录 - 可以提高转分化外周血细胞（PBMC）效率的因素 相关多巴胺能神经元。我们的中心假设是我们可以精确控制时间，剂量和 转录因子的组合以在A中创建高质量的功能性细胞产品 与当前方法相比，时间短，并且没有与病毒基因递送相关的风险。支持- 在这个目标上，我们已经证明了仅挤压治疗不会显着影响基因 表达，我们可以有效地从IPSC产生神经元，并且可以引入多个转录因子 进入PBMC，以上调关键神经元信号通路的表达。理由是我们的非病毒 传递转录因子以驱动细胞命运的方法可以显着提高效率和效率 与其他转变方法相比，与其他安全性和调节问题相比，产生的细胞的细胞生产。 此外，与同种异体IPSC衍生产品相比，自体细胞不需要慢性 免疫抑制 - 确保患者长期健康的关键因素。在AIM 1中，我们试图优化 细胞挤压技术的方法将基于mRNA的转录因子传递到PBMC以驱动努力 - 将良性转分解为多巴胺能神经元（DNS）。最终的DNS将在 体外，与使用现有方法从IPSC生成的DN进行了比较。在AIM 2中，这些DN将在功能上是 在体内鼠PD模型中评估，以支持进一步发展为潜在变化细胞 治疗。这些目标的成功完成可能会支持与其他生物制药的合作机会 在神经退行性中寻求分化的细胞疗法方法的公司。