Develop novel assays for assessing cellular and gene therapies

开发评估细胞和基因疗法的新方法

• 批准号: 8565299
• 负责人: Harvey Klein
• 金额: --
• 依托单位: CLINICAL CENTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8565299
• 关键词: B-Lymphocytes; Biological Assay; Biological Markers; Bone Marrow; Cell Culture Techniques; Cell Therapy; Cell physiology; Cells; Clinical; Clinical effectiveness; Complex; Dendritic Cells; Engraftment; Enzyme-Linked Immunosorbent Assay; Gene Expression; Gene Expression Profiling; Genes; Genetic Transcription; Hematopoietic stem cells; Laboratories; Leukocytes; Measures; MicroRNAs; Molecular Profiling; Outcome; Outcome Measure; Population; Production; Recurrent disease; Sirolimus; Stem cell transplant; Stromal Cells; T-Lymphocyte; Testing; Time; Transplant Recipients; Unmarried person; enzyme linked immunospot assay; gene therapy; improved; monocyte; novel; peripheral blood; potency testing; prevent; stem cells; sterility testing

Cell and gene therapy products must be tested for sterility, stability, purity and potency. In addition, its important to test clinical cell thearpy products for identity, consistency and comparability. Testing cellular and gene therapies is challenging. These therapies are generally collected from a single person so the quantity of material available to test is limited. They are typically transfused immediately after they are produced so there is a very limited amount of time to complete the assays. Many of these therapies are complex cells that have multiple functions. The cell functions that are critical to the clinical effectiveness of these therapies are often not known. Traditionally, analytic assays such as flow cytometery, ELISA, ELISPOT and cell culture have been used to analyze cellular and gene therapies. While these assays have proven to be very useful, the number and types of factors that can be analyzed with these assays is limited. We have been investigating the use of gene and micro RNA expression assays for the analysis of cellular therapies. These assays can require the use of only small quantities of cells and can be used to test up to 36,000 factors at one time. We have been testing the ability of global gene and micro RNA expression profiling to determine the utility of these assays for assessing the stability, purity and potency of cellular therapies. We have shown that gene expression profiling can detect changes in stored cells and detect differences between peripheral blood leukocytes (T cells, B cells and monocytes) and hematopoietic stem cells. Gene expression profiling has also been able to detect differences between immature and mature dendritic cells (DCs) and has been useful for comparing mature DCs produced using different combinations of maturation agents. We will soon begin analyzing clinical DC products to compare gene expression with clinical outcome in order to identify biomarkers that might be useful of consistency, stability, identity and potency testing. We have assessed global gene expression profiles of Th2 Rapamycin cells that are given to hematopoietic stem cell transplant patients to improve stem cell engraftment and prevent disease relapse. The studies have revealed several biomarkers which by be useful for identity, consistency and potency testing of these products. We are now comparing Th2 Rapamycin product biomarkers with clinical outcome measure to identify potency biomarkers. We have been investigating sensecence of bone marrow stromal cells (BMSCs) and have found a 21 gene set that predicts BMSC time in culture as measured by population doublings.

细胞和基因治疗产品必须进行无菌、稳定性、纯度和效力测试。此外，对临床细胞学产品的一致性、一致性和可比性进行测试也很重要。测试细胞和基因疗法是具有挑战性的。这些疗法通常是从一个人身上收集的，因此可供测试的材料数量有限。它们通常在生产后立即输注，因此完成检测的时间非常有限。这些疗法中的许多都是具有多种功能的复杂细胞。对这些疗法的临床效果至关重要的细胞功能通常是未知的。传统的分析方法有流式细胞术、酶联免疫吸附试验、ELISPOT和细胞培养等，用于细胞和基因治疗的分析。虽然这些化验已被证明非常有用，但可以用这些化验分析的因素的数量和类型是有限的。 我们一直在研究使用基因和微型RNA表达分析来分析细胞疗法。这些分析只需要使用少量的细胞，一次可以测试多达36,000个因素。我们一直在测试全球基因和微RNA表达谱分析的能力，以确定这些分析在评估细胞治疗的稳定性、纯度和有效性方面的效用。我们已经证明，基因表达谱可以检测储存细胞的变化，并检测外周血白细胞(T细胞、B细胞和单核细胞)和造血干细胞之间的差异。基因表达谱也能够检测未成熟和成熟树突状细胞之间的差异，并用于比较不同成熟剂组合产生的成熟树突状细胞。我们将很快开始分析临床DC产品，将基因表达与临床结果进行比较，以确定可能对一致性、稳定性、身份和效力测试有用的生物标记物。 我们评估了给予造血干细胞移植患者的Th2雷帕霉素细胞的全球基因表达谱，以改善干细胞植入和防止疾病复发。研究揭示了几种生物标志物，它们对这些产品的身份、一致性和效力测试很有用。我们现在正在比较Th2雷帕霉素产品生物标记物与临床结果指标，以确定效力生物标记物。 我们一直在研究骨髓基质细胞(BMSCs)的感觉，并发现了一个21基因集，通过群体倍增来预测BMSC在培养中的时间。