Development of a Mouse Model to Study Targeted Therapy to Prevent CNS Invasion by Pediatric T-ALL

开发小鼠模型来研究预防儿童 T-ALL 侵袭中枢神经系统的靶向治疗

基本信息

批准号：
10579626
负责人：
CHARLOTTE M VINES
金额：
$ 7.68万
依托单位：
UNIVERSITY OF TEXAS EL PASO
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-15 至 2024-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10579626
关键词：
Acute Lymphocytic Leukemia Acute T Cell Leukemia Animals Blood Blood - brain barrier anatomy Breeding CC chemokine receptor 7 CCL19 gene Cells Central Nervous System Central Nervous System Leukemia Cephalic Child Childhood Acute Lymphocytic Leukemia Childhood Precursor T Lymphoblastic Leukemia Circulation Clinical Complication Cranial Irradiation Data Development Diagnosis Enterobacteria phage P1 Cre recombinase Flow Cytometry Generations Goals Growth Human Immune Immune system Intelligence Interferons Invaded Justice Knock-out Knockout Mice Leukemic Cell Ligands LoxP-flanked allele Luciferases Lymphatic Macrophage Malignant Childhood Neoplasm Malignant Neoplasms Manuscripts Mediating Meninges Modeling Molecular Mouse Strains Mus NOTCH1 gene Nervous System Trauma Organ Organism Outcome Patients Peptide Receptor Peptides Peripheral Physicians Publishing Regimen Reporting Research Design Research Personnel Role Second Primary Cancers Signal Transduction Site T-Lymphocyte Testing Therapeutic Therapeutic Uses Tumor Cell Invasion Tumor-infiltrating immune cells Visualization Xenograft procedure acute T-cell lymphoblastic leukemia cell antagonist cell type chemokine receptor chemotherapy conditional knockout falls glymphatic system in vivo leukemia leukemia relapse leukemogenesis lymph nodes migration mouse model novel overexpression pediatric patients prevent receptor relapse prevention response side effect sphingosine 1-phosphate targeted treatment tool trafficking transcription factor

项目摘要

TITLE: Development of a Mouse model to test targeted therapy to prevent CNS Invasion by Pediatric T- ALL Abstract Leukemia is the most frequent pediatric cancer with acute lymphoblastic leukemia (ALL) being the most common leukemia type in children. In 15-20% of pediatric T-cell ALL (T-ALL) patients, the cancer invades the Central Nervous System (CNS), a site which is protected from systemic chemotherapies by the blood brain barrier. The invading leukemic cells are reservoirs, which can emerge and re-enter the circulation following treatment and cause T-ALL relapse. To prevent relapse, pediatric patients are treated with cranial irradiation and toxic chemotherapies, which are injected directly into the CNS. It is not surprising that these treatments can lead to morbid life-long side effects such as reduced intelligence, stunted growth, and secondary cancers in the CNS. Generally, the T-ALL has entered the CNS prior to diagnosis, and it is unclear if blocking the entry of T-ALL into the CNS at that point, after they have already entered, provides any therapeutic benefit. The overarching three goals of this proposal are: 1) to develop a mouse model of T- ALL that recapitulates human T-ALL, in which we can see the cells in living animals 2) to determine if blocking T-ALL cells from entering the CNS, prevents further T-ALL CNS invasion and 3) to determine if the T-ALL that has invaded the CNS will eventually clear the CNS and return to the circulation. Our long-term goal is to reduce or eliminate the need for damaging intrathecal or cranial treatments in patients with T-ALL. Our studies are based on a human xenograft mouse model of T-ALL, which revealed that during leukemogenesis, expression of NOTCH1, a transcription factor which is expressed in more than 80% of pediatric T-ALLs, induces expression of CC chemokine receptor 7 (CCR7). The study revealed that activation of CCR7 in T-ALL by one of its ligands, CCL19 promotes chemotactic migration of T-ALL into the CNS. Our proposal is based on a mouse model obtained from Dr. Monica Justice which uses an inducible CCR7(+)ROSA26 floxed-stop-PRDM14 (R26PR) locus. When crossed to an MX1-Cre these R26PR mice develop leukemia in response to overexpression of NOTCH1, which invades the CNS. Subsequent breeding of these mice with ROSA26 floxed-stop luciferase mice will result in Cre-recombinase induced expression of bioluminescent luciferase in T-ALL cells. To confirm a role for CCR7 in the invasion of the CNS, these R26PR mice will be crossed with our own CCR7 conditional knockout mice (CCR7floxed/floxed (CCR7f/f)) to produce R26PRDM14/CCR7f/f/MX1-CRE mice. Generation of this mouse is aim 1. Determining which immune cells co-localize with T-ALL cells upon entry into the CNS is aim2 and determining if blocking CCR7 using the CCL19 antagonist, CCL198-83 prevents or significantly reduces T-ALL CNS invasion, clears the CNS of T-ALL and increases mouse survival is aim 3. Ultimately, if successful, this mouse model will provide a tool for developing novel peptide therapies that can block T-ALL CNS invasion, without causing long-term CNS damage in patients and to study mechanisms of CNS invasion.

标题：开发小鼠模型来测试预防小儿 T- 侵袭中枢神经系统的靶向治疗全部抽象的白血病是最常见的儿科癌症，其中最常见的是急性淋巴细胞白血病 (ALL) 儿童白血病类型。在 15-20% 的儿童 T 细胞 ALL (T-ALL) 患者中，癌症侵犯中枢神经系统（CNS），一个受血脑屏障保护免受全身化疗的部位。白血病的侵袭细胞是储存库，在治疗后可能出现并重新进入循环系统，导致 T-ALL 复发。到为了防止复发，儿科患者接受颅脑照射和注射有毒化疗药物进行治疗直接进入中枢神经系统。毫不奇怪，这些治疗可能会导致病态的终生副作用，例如减少智力、发育迟缓和中枢神经系统继发性癌症。一般来说，T-ALL 在进入 CNS 之前诊断，目前尚不清楚在 T-ALL 已经进入中枢神经系统后是否会阻止它们进入中枢神经系统，提供任何治疗益处。该提案的三个首要目标是：1）开发 T-小鼠模型 ALL 概括了人类 T-ALL，其中我们可以看到活体动物中的细胞 2) 以确定是否阻断 T-ALL 细胞进入中枢神经系统，防止 T-ALL 进一步侵入中枢神经系统，3) 确定 T-ALL 是否已侵入中枢神经系统 CNS最终会清除CNS并返回循环。我们的长期目标是减少或消除这种需求用于 T-ALL 患者的损伤性鞘内或颅内治疗。我们的研究基于人类异种移植 T-ALL 小鼠模型，该模型揭示了在白血病发生过程中，NOTCH1（一种转录因子）的表达在超过 80% 的儿童 T-ALL 中表达，诱导 CC 趋化因子受体 7 (CCR7) 的表达。研究揭示了 T-ALL 中的 CCR7 通过其配体之一的激活，CCL19 促进 T-ALL 趋化迁移到中枢神经系统。我们的建议基于从 Monica Justice 博士获得的小鼠模型，该模型使用诱导型 CCR7(+)ROSA26 floxed-stop-PRDM14 (R26PR) 基因座。当与 MX1-Cre 杂交时，这些 R26PR 小鼠会因响应而患上白血病 NOTCH1 过度表达，侵入中枢神经系统。随后用 ROSA26 floxed-stop 培育这些小鼠荧光素酶小鼠将导致 Cre 重组酶诱导 T-ALL 细胞中生物发光荧光素酶的表达。确认由于 CCR7 在 CNS 侵袭中的作用，这些 R26PR 小鼠将与我们自己的 CCR7 条件敲除小鼠杂交小鼠（CCR7floxed/floxed (CCR7f/f)）产生 R26PRDM14/CCR7f/f/MX1-CRE 小鼠。该鼠标的生成是目标 1。目标2是确定哪些免疫细胞在进入中枢神经系统后与 T-ALL 细胞共定位，并确定是否阻断 CCR7使用CCL19拮抗剂CCL198-83预防或显着减少T-ALL CNS侵袭，清除T-ALL的CNS 目标 3 是提高小鼠的存活率。最终，如果成功，该小鼠模型将为开发新型小鼠提供工具可以阻断 T-ALL CNS 侵袭且不会对患者造成长期 CNS 损伤的肽疗法正在研究中枢神经系统侵袭机制。