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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：对 CD19 具有特异性的临床级 T 细胞已表现出抗肿瘤活性，我们现在提出一项转化研究，以研究与过继转移的 CD19 特异性 T 细胞相关的时空生物分布和微环境。为了针对侵袭性 B 细胞恶性肿瘤，我们启动了两项临床试验，以注入经过基因改造的自体和同种异体 T 细胞。表达 CD19 特异性嵌合抗原受体 (CAR)，该受体可识别细胞表面的 CD19，独立于 MHC。这项新的 R01 拨款申请建立了跨学科（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（CellSight Technologies, Inc.）合作，研究通过 PET 对输注的 CAR+ T 细胞进行成像的平台。可以通过使用睡美人 (SB) 转座子/转座酶系统在 T 细胞中共表达单纯疱疹病毒 1 型胸苷激酶 (sr39tk) 突变体和 CD19 特异性 CAR 来完成，我们已将该系统用于临床转化。使用我们称为 ¿ 的新方法转移表达 SB 转座子 (i) CAR 和 (ii) sr39tk 的两个 DNA 质粒双换位¿目标 #1 旨在确定非病毒基因转移是否会产生在组成型和条件启动子控制下共表达 CD19 特异性 CAR 和 sr39tk 的 T 细胞。 sr39tk 报告基因将与潮霉素磷酸转移酶 (Hy) 融合，从而与 CAR 融合。 +sr39tk+ T 细胞将在存在杀细胞浓度的潮霉素 B 的情况下选择性增殖γ-照射的人工抗原呈递细胞与所需的 T 细胞共刺激分子共表达 CD19，目标#2 旨在对免疫受损的人 CAR+sr39tk+ T 细胞进行纵向 μPET 成像。通过比较 (i) NFAT 控制下 sr39tk 的条件表达，对小鼠评估 T 细胞激活状态的生物分布和敏感性。 (ii) CellSight 开发的新型 PET 探针 [18F]F-AraG 将通过引入氧分子传感器来评估 sr39tk 是否可以报告低氧张力。一项新临床研究的临床前数据，该研究将 CAR+sr39tk+ T 细胞注入接受 CD19 特异性 T 细胞基因治疗的患者中。该试验将是我们现有试验的配套研究。 (IND# 14193) 在自体造血干细胞移植后为患有晚期 B 淋巴恶性肿瘤的研究参与者输注 CAR+ T 细胞由 CellSight Technologies 销售的 PET 探针 [18F]FHBG 将根据 IND 在 TMH 制造用于临床成像。 #61880 总的来说，这些研究将检验 CD19 特异性 CAR+sr39tk+ T 细胞可以被这些研究将建立基于 PET 的 CAR+ T 细胞成像的原理和实践，并提供有关转基因 T 细胞生物分布的第一个人体成像数据。