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Principal Investigator: Juliane Gust
Organization: SEATTLE CHILDREN'S HOSPITAL
Fiscal Year: 2024
Award: $549,663
Funding agency: National Cancer Institute
PROJECT SUMMARY
This project studies the mechanism of neurologic toxicity in chimeric antigen receptor (CAR) T cell
therapy. CAR T cells are genetically modified, patient derived T cells that use the CAR to recognize and destroy
malignant target cells. Although CAR T therapy has shown impressive results against leukemia and lymphoma,
approximately 30-40% of patients experience neurologic side effects in the first month after receiving CD19-
targeted CAR T cells. This includes cognitive disturbances, seizures, and in rare cases fatal cerebral edema.
Systemic cytokine release syndrome after CAR T cell infusion is a well-established risk factor for neurotoxicity,
but the connection between systemic inflammation and brain dysfunction is poorly understood.
To study the mechanisms of neurotoxicity, we have developed an immunocompetent mouse model. After
treatment with high dose CD19-directed murine CAR T cells, mice develop motor and balance difficulties, as
well as brain microhemorrhages. Surprisingly, we found that >10% of cortical capillaries are obstructed by white
blood cells during neurotoxicity. This was accompanied by capillary remodeling and decreased vessel coverage
by pericytes. Based on these findings, we now propose the following experiments:
Aim 1: What molecular mechanisms cause white blood cells to plug capillaries during neurotoxicity? We
will use in vivo two-photon imaging in mice to determine which cell types cause the capillary plugging – the
mouse’s own or the transferred CAR T cells? We will then measure how CAR T cell treatment changes the
expression of adhesion molecules in brain capillary endothelial cells and in leukocytes, and test whether
blockade of these adhesion interactions can prevent capillary plugging and neurotoxicity.
Aim 2: Is neuroendothelial-leukocyte adhesion increase in human microvessels during neurotoxicity? In
parallel to our work in mice, we will use a 3D in vitro model of human brain capillaries to measure how soluble
factors in patient plasma affect adhesion molecule expression in the endothelium. We will then test whether
white blood cells from CAR T cell patients with neurotoxicity have increased predilection for plugging synthetic
microvessels that mimic capillaries, and whether we can prevent this plugging by blocking adhesion molecules.
Aim 3: Can the strength of cell-cell adhesion signaling separate CAR T cell efficacy from toxicity? We will
use quantitative multiplex immunoprecipitation to probe protein-protein interaction networks in CAR T cells that
cause high or low neurotoxicity in mice to understand what activation states are conducive to neurotoxicity. We
will then test whether knock down of adhesion molecule expression can direct CAR T cells away from a toxicity
phenotype by impairing their ability to signal to other cells, and to adhere to the brain microvasculature.
This work is innovative because it combines advanced imaging, in vitro modeling techniques, and protein
network analysis in a unique collaboration between neuroscience, vascular biology, and oncology. The work is
significant because it addresses key safety issues in emerging cancer immunotherapy modalities.
Terms: <0-11 years old><3-D><3-Dimensional><3D><Ablation><Acute><Address><Adhesion Molecule><Adhesions><Adventitial Cell><Affect><After Care><After-Treatment><Aftercare><Animal Model><Animal Models and Related Studies><Antibodies><Antigenic Determinants><Basic Research><Basic Science><Behavior><Behavioral><Binding Determinants><Biology><Biomedical Engineering><Bleeding><Blood Plasma><Blood Vessels><Blood capillaries><Blood leukocyte><Brain><Brain Dead><Brain Death><Brain Edema><Brain Nervous System><Brain Swelling><CAR T cell therapy><CAR T cells><CAR T therapy><CAR modified T cells><CAR-T><CAR-Ts><CD19><CD19 gene><CNS Nervous System><Cancers><Capillary Endothelial Cell><Cell Adhesion Molecule Gene><Cell Adhesion Molecules><Cell Body><Cell Communication and Signaling><Cell Signaling><Cell-Cell Adhesion><Cells><Cellular immunotherapy><Central Nervous System><Cerebral Edema><Child><Child Youth><Childhood><Children (0-21)><Clinical><Clinical Trials><Cognitive Disturbance><Cognitive Impairment><Cognitive decline><Cognitive function abnormal><Collaborations><Collagen><Coma><Coma Depasse><Comatose><Delirium><Development><Disturbance in cognition><Dose><Encephalon><Endothelium><Engineering><Epitopes><Equilibrium><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Gene Expression Monitoring><Gene Expression Pattern Analysis><Gene Expression Profiling><Genetic><Hematologic Cancer><Hematologic Malignancies><Hematologic Neoplasms><Hematological Malignancies><Hematological Neoplasms><Hematological Tumor><Hematopoietic Cancer><Hemorrhage><Human><Human Engineering><Hydrogels><Hypoxia><Hypoxic><Image><Immune Precipitation><Immune system><Immunocompetent><Immunoprecipitation><Impaired cognition><Impairment><In Vitro><Inflammatory><Infusion><Infusion procedures><Injury><Integrin Binding><Integrins><Integrins Extracellular Matrix><Interdisciplinary Research><Interdisciplinary Study><International><Intracellular Communication and Signaling><Intracranial Edema><Investigation><Leukocytes><Leukocytes Reticuloendothelial System><Link><Malignant><Malignant - descriptor><Malignant Cell><Malignant Hematologic Neoplasm><Malignant Neoplasms><Malignant Tumor><Marrow leukocyte><Measures><Mediating><Medical><Mice><Mice Mammals><Modality><Modeling><Modern Man><Molecular><Motor><Multidisciplinary Collaboration><Multidisciplinary Research><Murine><Mus><Network Analysis><Neuraxis><Neurologic><Neurological><Neuronal Dysfunction><Neurosciences><Oncology><Oncology Cancer><Oxygen Deficiency><Pathway Analysis><Patients><Perfusion><Pericapillary Cell><Pericytes><Perivascular Cell><Phenotype><Plasma><Plasma Serum><Position><Positioning Attribute><Preventative intervention><Property><Proteins><Protocol><Protocols documentation><Receptor Activation><Reticuloendothelial System, Serum, Plasma><Risk><Risk Factors><Rouget Cells><Safety><Sampling><Seizures><Short interfering RNA><Signal Induction><Signal Transduction><Signal Transduction Systems><Signaling><Small Interfering RNA><Stimulus><T cells for CAR><T-Cell Proliferation><T-Cell Receptor Therapy><T-Cell Receptor Treatment><T-Cell Receptor based Therapy><T-Cell Receptor based Treatment><T-Cells><T-Lymphocyte><TCR Therapy><TCR based Therapy><TCR based treatment><Techniques><Testing><Therapeutic Intervention><Toxic effect><Toxicities><Transcript Expression Analyses><Transcript Expression Analysis><Transgenic Organisms><Translating><White Blood Cells><White Cell><Work><abnormal brain function><analyze gene expression><anti-cancer immunotherapy><anticancer immunotherapy><balance><balance function><bio-engineered><bio-engineers><bioengineering><biological engineering><biological signal transduction><blood loss><brain capillary><brain dysfunction><brain endothelial cell><brain impairment><brain microvascular endothelial cell><brain microvasculature><brain microvessels><brain vascular endothelial cell><cancer cell><cancer immunotherapy><capillary><cell adhesion protein><cell type><cell-based immunotherapy><cerebral capillary><cerebral death><cerebral endothelial cell><cerebral microvascular endothelial cell><cerebral microvasculature><cerebral microvessels><cerebral vascular endothelial cell><chimeric antigen T cell receptor><chimeric antigen receptor><chimeric antigen receptor (CAR) T cell therapy><chimeric antigen receptor (CAR) T cells><chimeric antigen receptor T cell therapy><chimeric antigen receptor T cells><chimeric antigen receptor T therapy><chimeric antigen receptor fusion protein T-cells><chimeric antigen receptor modified T cells><cognitive dysfunction><cognitive loss><cohort><cytokine><cytokine release syndrome><cytokine storm><delirious><developmental><dysfunctional brain><experience><experiment><experimental research><experimental study><experiments><flow cytophotometry><gene expression analysis><gene expression assay><genetically engineered cells><genetically modified cells><high risk><human model><imaging><imaging in vivo><immune cell therapy><immune competent><immune-based cancer therapies><immunological synapse><immunotherapy for cancer><immunotherapy of cancer><in vitro Model><in vivo><in vivo imaging><in vivo two-photon imaging><infusions><injuries><innovate><innovation><innovative><integrin bound><intervention for prevention><intervention therapy><kids><knock-down><knockdown><leukemia/lymphoma><lymphoma/leukemia><malignancy><manufacture><model of animal><model of human><mouse model><murine model><natalizumab><neoplasm/cancer><neural dysfunction><neuro-vascular unit><neuron toxicity><neuronal toxicity><neurotoxicity><neurovascular unit><novel><pediatric><peripheral blood><post treatment><pre-clinical safety><preclinical safety><prevent><preventing><prevention intervention><preventional intervention strategy><preventive intervention><protein protein interaction><receptor binding><receptor bound><siRNA><side effect><systemic inflammation><systemic inflammatory response><three dimensional><thymus derived lymphocyte><transcriptional profiling><transgenic><tumor><vascular><wet brain><white blood cell><white blood corpuscle><youngster>