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Blood Brain Barrier Disruption during Chimeric Antigen Receptor (CAR) T cell therapy

嵌合抗原受体 (CAR) T 细胞治疗期间的血脑屏障破坏

基本信息

批准号：
10249232
负责人：
Juliane Gust
金额：
$ 18.74万
依托单位：
SEATTLE CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-01 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10249232
关键词：
Address Adherence Adhesions Affect Animal Model Animals Area Astrocytes Award Basement membrane Behavior Biology Blood Blood - brain barrier anatomy Blood Platelets Blood Vessels Blood capillaries Blood flow Brain Brain Edema CAR T cell therapy CD19 gene Caliber Cell Adhesion Cell Count Cell surface Cells Cellular immunotherapy Cephalic Cerebral Edema Cerebrovascular Circulation Cerebrum Chemotherapy and/or radiation Child Health Clinical Communication Control Groups Data Delirium Dextrans Dose Edema Electroencephalography Electron Microscopy Endothelial Cells Endothelium Erythrocytes Experimental Designs Extravasation Flow Cytometry Functional disorder Goals Hematologic Neoplasms Hemorrhage Histology Human Image Imaging Techniques Immunology Immunotherapy Impaired cognition Impairment Inbred Strain Individual Infusion procedures Injury Knowledge Label Leadership Leukemic Cell Leukocytes Life Link Lymphoma Malignant Neoplasms Measures Mechanics Mentors Metalloproteases Modeling Molecular Mus Neurologic Neurologic Dysfunctions Neurology Oncology Pathway interactions Patient Care Patients Perfusion Pericytes Preparation Program Development Proliferating Proteins Refractory Relapse Research Resolution Risk Risk Factors Science Scientist Seizures Severities Shapes Speed Structure Surface Syndrome T-Lymphocyte Testing Therapeutic Thinness Tight Junctions Toxic effect Tracer Training Tumor-infiltrating immune cells Vesicle Visualization Wild Type Mouse Work animal model development arteriole blood-brain barrier disruption brain dysfunction brain health cancer cell cancer immunotherapy career career development cell killing cell type cerebral microbleeds cerebrovascular chemotherapy chimeric antigen receptor chimeric antigen receptor T cells cognitive change common symptom cranium cytokine cytokine release syndrome experimental analysis hemodynamics hypoperfusion improved in vivo in vivo two-photon imaging innovation interstitial leukemia/lymphoma model development mouse model neurotoxicity neurovascular unit novel postcapillary venule prevent receptor rhodamine 6G skills systemic inflammatory response therapeutic development translational physician vasoconstriction venule young adult

项目摘要

PROJECT SUMMARY/ABSTRACT Dr. Juliane Gust proposes a study to understand whether perturbations of the BBB and cerebral microvascular perfusion contribute to CAR T cell neurotoxicity. This work will prepare Dr. Gust for independence as a translational clinician-scientist at the intersection of neurology, oncology, and immunology. In CAR T cell therapy, patients’ T cells are modified with a receptor that recognizes cancer cell surface markers, and induces T cell killing of the target. Thousands of patients with previously little hope of cure have benefitted from CD19-directed CAR T cells for leukemia and lymphoma. However, ~40% develop neurologic toxicity, and ~1% die from cerebral edema. The mechanism of neurotoxicity is poorly understood. In patients, Dr. Gust has shown evidence of endothelial activation, glial injury, leukocyte infiltrates, and microhemorrhages. To model neurotoxicity in mice, Dr. Gust treated wild type mice with high dose CD19-CAR T cells made from syngeneic donor mice of the same inbred strain. CAR T treated mice, unlike mice treated with untransduced T cells, develop systemic cytokine release, abnormal behavior, and widespread cerebral microhemorrhages. Taken together, the human and mouse data suggest the following hypotheses: that the BBB is disrupted during neurotoxicity (Aim 1), and that neurotoxicity is accompanied by altered cerebral blood flow (Aim 2). For Aim 1, Dr. Gust will use immunolabeling of individual NVU components (endothelial cells, tight junctions, pericytes, basement membrane, astrocyte endfeet) and quantify cell number, shape, and contiguity. She will inject intravascular tracers followed by fluorescent and electron microscopy to visualize tracer leakage via paracellular and transcellular pathways. She will assess the contribution of immune infiltrates to BBB breakdown by flow cytometry and histology with colabeling for matrix metalloprotease-9. If disruption of the NVU by structural or functional alteration is confirmed, we can conclude that it is a key link from systemic inflammation to brain dysfunction, which warrants further detailed mechanistic studies. For Aim 2, Dr. Gust will measure blood flow in mouse cortical arterioles, capillaries and venules via in-vivo two-photon imaging through a thinned skull window. This innovative approach allows visualization of hemodynamics with single microvessel resolution by measuring vessel diameter and speed of red blood cell transit. To test the hypothesis that CAR T cell treatment leads to leukocyte adherence to vessel walls and consequent slowing of blood flow, transit and rolling of GFP-expressing CAR T cells and Rhodamine-6G labeled leukocytes and platelets will be quantified. Confirmation of impaired microvascular blood flow would guide a reconsideration of therapeutic approaches. During the award period, Dr. Gust will receive training in animal model development, advanced imaging techniques, immunology, vascular biology, rigor in experimental design and analysis, science communication, networking, and leadership skills. Under the guidance of primary mentor Dr. Andy Shih and her mentoring team, she will use data from this proposal to develop an R01 application for transition to independence.

项目总结/文摘