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: 8952807
• 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/8952807
• 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; Peptides; Population; Production; Recurrent disease; Sirolimus; Stem cell transplant; Stem cells; Stromal Cells; T-Lymphocyte; Testing; Time; Transplant Recipients; Unmarried person; enzyme linked immunospot assay; gene therapy; improved; monocyte; novel; peripheral blood; potency testing; prevent; 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 44,000 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 T 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 may be useful for identity, consistency and potency testing of these products. We are now comparing T 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 24 gene set that predicts BMSC time in culture as measured by population doublings. We have also been seraching for biomarkers expressed by peptide-plused mature dendrtic cells whose expression coorelates with clinical effectiveness.

细胞和基因治疗产品必须经过无菌性、稳定性、纯度和效力测试。此外，对临床细胞治疗产品进行鉴定、一致性和可比性也很重要。测试细胞和基因疗法具有挑战性。这些疗法通常是从一个人身上收集的，因此可用于测试的材料数量有限。它们通常在生产后立即输注，因此完成测定的时间非常有限。这些疗法中有许多是具有多种功能的复杂细胞。对这些治疗的临床效果至关重要的细胞功能通常不为人所知。传统上，流式细胞仪、ELISA、ELISPOT和细胞培养等分析方法已被用于分析细胞和基因治疗。虽然这些分析已被证明是非常有用的，但可以用这些分析分析的因素的数量和类型是有限的。