IMAGING T CELLS BY POSITRON EMISSION TOMOGRAPHY

通过正电子发射断层扫描对 T 细胞进行成像

• 批准号: 8539750
• 负责人: Laurence J.N. Cooper
• 金额: $ 57.51万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-04 至 2017-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8539750
• 关键词: Academia; Allogenic; Antigen Receptors; Antigen-Presenting Cells; Applications Grants; Architecture; Area; Autologous; B lymphoid malignancy; Biodistribution; Bioinformatics; Biometry; CD19 gene; Cell Hypoxia; Cell surface; Chemistry; Clinical; Clinical Data; Clinical Research; Clinical Trials; Companions; Cyclotrons; Cytoplasm; Data; Detection; Discipline of Nuclear Medicine; Electroporation; Enrollment; Event; Exhibits; Firefly Luciferases; Gene Transfer; Goals; Guanine; Hematopoietic Stem Cell Transplantation; Herpesvirus 1; Human; Hypoxia; Image; Immunocompromised Host; Immunology; In Vitro; Industry; Infusion procedures; Institution; Instruction; Lymphoma; Marketing; Measurement; Methods; Modification; Molecular; Monitor; Monkeys; Mus; Oxygen; Oxygen measurement, partial pressure, arterial; Participant; Patients; Phase I Clinical Trials; Positron-Emission Tomography; Renilla Luciferases; Reporter; Reporter Genes; Reporting; Research; Research Personnel; Sleeping Beauty; Stress; System; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Technology; Testing; Thymidine Kinase; Transgenes; Translating; Translations; Transposase; University of Texas M D Anderson Cancer Center; base; bioluminescence imaging; cell bank; clinical application; gene therapy; hygromycin A; hygromycin-B kinase; improved; in vivo; mutant; open source; peripheral blood; plasmid DNA; pre-clinical; promoter; sensor; tool; translational study; tumor; tumor microenvironment; uptake; Academia; Allogenic; Antigen Receptors; Antigen-Presenting Cells; Applications Grants; Architecture; Area; Autologous; B lymphoid malignancy; Biodistribution; Bioinformatics; Biometry; CD19 gene; Cell Hypoxia; Cell surface; Chemistry; Clinical; Clinical Data; Clinical Research; Clinical Trials; Companions; Cyclotrons; Cytoplasm; Data; Detection; Discipline of Nuclear Medicine; Electroporation; Enrollment; Event; Exhibits; Firefly Luciferases; Gene Transfer; Goals; Guanine; Hematopoietic Stem Cell Transplantation; Herpesvirus 1; Human; Hypoxia; Image; Immunocompromised Host; Immunology; In Vitro; Industry; Infusion procedures; Institution; Instruction; Lymphoma; Marketing; Measurement; Methods; Modification; Molecular; Monitor; Monkeys; Mus; Oxygen; Oxygen measurement, partial pressure, arterial; Participant; Patients; Phase I Clinical Trials; Positron-Emission Tomography; Renilla Luciferases; Reporter; Reporter Genes; Reporting; Research; Research Personnel; Sleeping Beauty; Stress; System; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Technology; Testing; Thymidine Kinase; Transgenes; Translating; Translations; Transposase; University of Texas M D Anderson Cancer Center; base; bioluminescence imaging; cell bank; clinical application; gene therapy; hygromycin A; hygromycin-B kinase; improved; in vivo; mutant; open source; peripheral blood; plasmid DNA; pre-clinical; promoter; sensor; tool; translational study; tumor; tumor microenvironment; uptake

DESCRIPTION (provided by applicant): Clinical-grade T cells rendered specific for CD19 have demonstrated anti-tumor activity. We are now proposing a translational study to investigate the temporal-spatial biodistribution and microenvironment associated with adoptively transferred CD19-specific T cells as achieved using positron emission tomography (PET). To target aggressive B-cell malignancies, we have initiated two clinical trials to infuse autologous and allogeneic T cells that have been genetically modified to express a CD19-specific chimeric antigen receptor (CAR) which recognizes CD19 on the cell surface, independent of MHC. This new R01 grant application establishes an inter-disciplinary (chemistry, imaging, biostatistics, bioinformatics, nuclear medicine, gene therapy, and immunology) and multi-institution (MDACC and TMH) team, partnering with industry (CellSight Technologies, Inc.) to investigate a platform for imaging infused CAR+ T cells by PET. This will be accomplished by coexpressing a mutant of herpes simplex virus-1 thymidine kinase (sr39tk) with the CD19-specific CAR in T cells using the Sleeping Beauty (SB) transposon/transposase system which we have adapted for clinical translation. We will synchronously electro-transfer two DNA plasmids expressing the SB transposons (i) CAR and (ii) sr39tk, using a new method we dub ¿double transposition¿. Aim #1 seeks to determine if non-viral gene transfer will produce T cells that co-express CD19-specific CAR and sr39tk under control of constitutive and conditional promoters. The sr39tk reporter gene will be fused to hygromycin phosphotransferase (Hy) and thus CAR+sr39tk+ T cells will be selectively propagated in presence of cytocidal concentration of hygromycin B on γ- irradiated artificial antigen presenting cells that co-express CD19 along with desired T-cell co-stimulatory molecules. Aim #2 seeks to undertake longitudinal μPET imaging of infused human CAR+sr39tk+ T cells with the reporter probe [18F]FHBG in immunocompromised mice to assess biodistribution and sensitivity of detection. T-cell activation status will be imaged by comparing (i) conditional expression of sr39tk under control of NFAT promoter with (ii) the new PET probe [18F]F-AraG developed at CellSight. T-cell hypoxia will be assessed by introducing a molecular sensor for oxygen to test whether sr39tk can report low oxygen tension. Aim #3 seeks to translate these pre-clinical data to a new clinical study infusing CAR+sr39tk+ T cells in patients undergoing gene therapy with CD19-specific T cells. This trial will be a companion study to our existing trial (IND# 14193) infusing CAR+ T cells after autologous hematopoietic stem-cell transplantation for research participants with advanced B-lymphoid malignancies. The PET probe [18F]FHBG, marketed by CellSight Technologies, will be manufactured for clinical imaging at TMH, per IND #61880. In aggregate, these studies will test the central hypothesis that CD19-specific CAR+sr39tk+ T cells can be imaged in humans using PET. These studies will establish principles and practices for translating PET-based imaging of CAR+ T cells and provide the first human imaging data on the biodistribution of genetically modified T cells.

描述（由适用提供）：针对CD19的临床级T细胞表现出抗肿瘤活性。现在，我们提出了一项翻译研究，以研究使用正电子发射断层扫描（PET）实现的与适当转移CD19特异性T细胞相关的临时空间生物分布和微环境。为了靶向侵略性的B细胞恶性肿瘤，我们已经开始了两项临床试验，以注入自体和同种异体T细胞，这些细胞通常经过修饰，以表达CD19特异性的嵌合抗原受体（CAR），该抗原受体（CAR）在细胞表面识别CD19，与MHC无关。这项新的R01赠款应用建立了一个跨学科（化学，成像，生物统计学，生物信息学，核医学，基因治疗和免疫学）和多机构（MDACC和TMH）团队，与行业（Cellight Technologies，Inc。）合作，以调查宠物受感染的CAR+ T细胞的平台。这将通过使用“睡美人（SB）转座子/转座酶系统”中的CD19特异性CAR与CD19特异性CAR与CD19特异性CAR共同表达单纯疱疹病毒-1胸苷激酶（SR39TK）的突变体来实现，我们已经适应了临床翻译。我们将同步电转移表达SB转座子（I）汽车和（ii）SR39TK的两个DNA质粒，使用一种新方法，我们dub�AIM＃1旨在确定非病毒基因转移会产生T细胞是否会产生T细胞是否会产生与CD19特定于CD19特异性CAR和SR39T的CD19特异性启动仪和条件促进仪和条件促进仪和构造的构造。 SR39TK报道基因将融合到湿霉素磷酸转移酶（HY）中，因此将在湿霉素B上在γ-辐照的人工抗原上伴有thematife cd19 cd19 cd19的thegrymycin b上选择CAR+ SR39TK+ T细胞在γ-辐照抗原上的细胞含量浓度的情况下进行传播。 AIM＃2旨在使用记者探针[18F] FHBG在免疫功能低下的小鼠中对注入的人类CAR+ SR39TK+ T细胞进行纵向μPET成像，以评估检测的生物分布和敏感性。通过比较（i）在NFAT启动子控制下SR39TK的条件表达与（ii）（ii）新的PET探针[18F] F-ARAG在Cellight中开发的新的PET探针[18F] F-ARAG。 T细胞缺氧将通过引入氧的分子传感器来测试SR39TK是否可以报告低氧张力来评估。 AIM＃3试图将这些临床前数据转化为一项新的临床研究，在接受CD19特异性T细胞基因治疗的患者中注入CAR+ SR39TK+ T细胞。该试验将是我们现有试验（IND＃14193）的一项伴侣研究，在自体造血干细胞移植后，对患有晚期B淋巴恶性肿瘤的研究参与者进行了自体造血干细胞移植。由Cellight Technologies销售的PET探针[18F] FHBG将在IND＃61880处制造用于TMH的临床成像。总体而言，这些研究将检验一个中心假设：CD19特异性CAR+ SR39TK+ T细胞可以使用PET在人类中成像。这些研究将建立用于翻译CAR+ T细胞基于PET的成像的原理和实践，并提供有关一般修饰T细胞生物分布的第一个人类成像数据。