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Principal Investigator: Curtis Harris
Organization: DIVISION OF BASIC SCIENCES - NCI
Fiscal Year: 2024
Award: $2,600,582
Funding agency: National Cancer Institute
In collaboration with Dr. Carl June (University of Pennsylvania), we have generated the d133p53a-armored CAR-T cells and revealed that they show superior anti-tumor activity compared with the control CAR-T cells (KIMRIAH(registered trademark) in clinical use) in co-culture and mouse injection models of Nalm6 pre-B leukemia cells. Importantly, d133p53a has significantly improved CAR-T cells derived from patients with chronic lymphocytic leukemia (CLL) who failed to respond to current CAR-T cell therapy. The RNA expression profiling and metabolic assays have suggested that the d133p53a-CAR-T cells are less vulnerable to cellular senescence and apoptosis, maintain genome stability, and have metabolic benefits in nutrition-competitive tumor microenvironment, which are all ideal phenotypes to overcome current obstacles in CAR-T cell therapy. Our recent publication (Roselle et al., PNAS 121: e2317735121, 2024) is a milestone toward clinical trials in non-responder and refractory CLL cases, as well as further improvement and modifications of the CAR-T cells for hard-to-treat solid tumors. We have generated the d133p53a transgenic mice to examine whether d133p53a has therapeutic effects in vivo for progeria pathologies and neurodegenerative diseases. The Cre recombinase-regulated inducible expression of d133p53a by tamoxifen injection has been confirmed in all organs and tissues examined, including aorta, heart, skin (major sites of progeria pathologies) and brain (neurodegenerative diseases). They are currently 12- to 15-month-old without any sign of increased tumorigenesis (in marked contrast to p53 knockout mice), being consistent with our previous human cell data that d133p53a is non-oncogenic and non-mutagenic. We have intiated a cross-breeding of the d133p53a transgenic mice with a progeria model mouse strain (LmnaG609G), which recapitulates Hutchinson-Gilford progeria syndrome. Our transgenic mice are also ready to be cross-bred with a mouse model of Alzheimer's disease and to be treated with an established protocol of brain irradiation or traumatic brain injury, which mimics brain radiotherapy-induced late cognitive impairment or chronic traumatic encephalopathy, respectively. Through high-throughput screening of small molecule libraries, repurposed drugs and natural products, we have identified two candidate drugs that can enhance the expression of d133p53a. We have confirmed that these drugs increase d133p53a protein levels and inhibit cellular senescence and associated secretory phenotype in human astrocytes and progeria patients-derived fibroblasts. These drugs have been injected into a human p53-knocked-in mouse strain (Hupki), which endogenously expresses this otherwise human/primate-specific d133p53a, to examine whether they are active in mice in vivo. Once confirmed, these drug-treated Hupki mice can be another model for in vivo therapeutic effects of d133p53a. We are also performing mechanistic studies to elucidate the molecular basis of their activity on d133p53a, including RNA expression profiling, proteome and metabolome analyses of the drug-treated cells and mice. We have pioneered a new area of p53 research by investigating cancer-associated mutant versions of d133p53a and p53beta. One of our new findings in this area is that a cancer-associated mutation of p53 (R273H) confers d133p53a (otherwise non-oncogenic and non-mutagenic, as mentioned above) with oncogenic properties (i.e., increased proliferation, migration and invasion, impaired DNA repair, and inhibition of cellular senescence and apoptosis) via the IL4I1/IDO1/AHR pathway in GBM cells (Joruiz et al., Cell Death Dis. 15: 454, 2024). We have also revealed that siRNA knockdown of a splicing factor SRSF3 upregulates p53beta and induces cellular senescence and apoptosis in both p53-wild-type and -mutant GBM and lung cancer cells. Aiming at an efficient and cancer-specific delivery of this tumor-suppressive SRSF3 siRNA, we in collaboration with Dr. Drew Weissman (University of Pennsylvania) have generated the lipid nanoparticles (LNPs) encapsulating SRSF3 siRNA and coated with a scFv antibody to cancer-specific cell surface proteins (e.g., ROR1 and IL13RA2). These LNPs have been confirmed to efficiently knock down SRSF3 and induce p53beta, cellular senescence and apoptosis in GBM and lung cancer cells in vitro. They will be tested in mouse in vivo models (e.g., intracranially injected human GBM xenografts) and examined for cancel cell senescence and apoptosis, in vivo distribution profile, cancel cell-specific targeting, and adverse effects (e.g., liver damage). [Publications] Roselle C, Horikawa I, Chen L, Kelly AR, Gonzales D, Da T, Wellhausen N, Rommel PC, Baker D, Suhoski M, Scholler J, O'Connor RS, Young RM, Harris CC, June CH. Enhancing chimeric antigen receptor T cell therapy by modulating the p53 signaling network with d133p53a. Proc. Natl. Acad. Sci. USA. 121: e2317735121, 2024. Joruiz SM, von Muhlinen N, Horikawa I, Gilbert MR, Harris CC. Distinct functions of wild-type and R273H mutant d133p53a differentially regulate glioblastoma aggressiveness and therapy-induced senescence.
Terms: <AD model><Adverse effects><Aging><Alzheimer's disease model><Antibodies><Antioncogene Protein p53><Aorta><Apoptosis><Apoptosis Pathway><Area><Assay><Astrocytes><Astrocytus><Astroglia><B-Cell CLL><B-Cell Chronic Lymphocytic Leukemia><B-Cell Chronic Lymphogenous Leukemia><B-Cell Chronic Lymphoid Leukemia><B-Cell Lymphocytic Leukemia><B-Lymphocytic Leukemia><Bioassay><Biological Assay><Body Tissues><Brain><Brain Nervous System><Brain Trauma><CAR T cell therapy><CAR T cells><CAR T therapy><CAR modified T cells><CAR-T><CAR-Ts><CD8 Cell><CD8 T cells><CD8 lymphocyte><CD8+ T cell><CD8+ T-Lymphocyte><CD8-Positive Lymphocytes><CD8-Positive T-Lymphocytes><CRE Recombinase><Cancers><Cell Aging><Cell Body><Cell Communication and Signaling><Cell Death><Cell Senescence><Cell Signaling><Cell Surface Proteins><Cells><Cellular Aging><Cellular Senescence><Cellular Tumor Antigen P53><Chronic B-Lymphocytic Leukemia><Chronic Lymphatic Leukemia><Chronic Lymphoblastic Leukemia><Chronic Lymphocytic Leukemia><Chronic Lymphogenous Leukemia><Clinical><Clinical Trials><Co-culture><Cocultivation><Coculture><Coculture Techniques><Cognitive Disturbance><Cognitive Impairment><Cognitive decline><Cognitive function abnormal><Collaborations><Cranial Irradiation><Crossbreeding><DNA Damage Repair><DNA Repair><Data><Degenerative Neurologic Disorders><Development><Disturbance in cognition><Drug Screening><Drugs><Encapsulated><Encephalon><Enterobacteria phage P1 Cre recombinase><Expression Profiling><Fibroblasts><Future><Gene Transcription><Generations><Genetic Alteration><Genetic Change><Genetic Hybridization><Genetic Transcription><Genetic defect><Genome Stability><Genomic Stability><Glioblastoma><Goals><Grade IV Astrocytic Neoplasm><Grade IV Astrocytic Tumor><Grade IV Astrocytoma><Heart><Heterograft><Heterologous Transplantation><High Throughput Assay><Human><Hutchinson-Gilford Disease><Hutchinson-Gilford Syndrome><IL-13Ra><IL13RA1><IL13RA1 gene><Impaired cognition><Impairment><In Vitro><Injections><Injury to Liver><Interleukin-13 Receptor Alpha><Interleukin-13 Receptor Alpha 1><Intracellular Communication and Signaling><Invaded><Isoforms><KI mice><KO mice><Knock-in Mouse><Knock-out Mice><Knockout Mice><Leukemic Cell><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Malignant Tumor of the Lung><Malignant neoplasm of lung><Mediating><Medication><Metabolic><Mice><Mice Mammals><Modeling><Modern Man><Modification><Molecular><Molecular Tumor Suppression><Mouse Strains><Murine><Mus><Mutation><NR4><NTRKR1><Natural Products><Nerve Degeneration><Nervous System Degenerative Diseases><Neural Degenerative Diseases><Neural degenerative Disorders><Neurodegenerative Diseases><Neurodegenerative Disorders><Neurologic Degenerative Conditions><Neuron Degeneration><Neurotrophic Tyrosine Kinase Receptor-Related 1><Normal Cell><Null Mouse><Oncogenesis><Oncogenic><Oncoprotein p53><Organ><P53><Pathology><Pathway interactions><Patients><Pennsylvania><Pharmaceutical Preparations><Phenotype><Phosphoprotein P53><Phosphoprotein pp53><Premature Senility Syndrome><Primates><Primates Mammals><Progeria><Programmed Cell Death><Proliferating><Property><Protein Isoforms><Protein TP53><Proteins><Proteome><Protocol><Protocols documentation><Publications><Pulmonary Cancer><Pulmonary malignant Neoplasm><RNA Expression><RNA Splicing><ROR1><ROR1 gene><Radiation therapy><Radiotherapeutics><Radiotherapy><Receptor Tyrosine Kinase-Like Orphan Receptor 1><Refractory><Replicative Senescence><Research><Scientific Publication><Short interfering RNA><Signal Transduction><Signal Transduction Systems><Signaling><Site><Skin><Small Interfering RNA><Solid Neoplasm><Solid Tumor><Splicing><Syndrome><T cells for CAR><T8 Cells><T8 Lymphocytes><TP53><TP53 gene><TRP53><Tamoxifen><Testing><Therapeutic><Therapeutic Effect><Tissues><Trademark><Transcription><Transgenic Mice><Translating><Traumatic Brain Injury><Traumatic encephalopathy><Tumor Protein p53><Tumor Protein p53 Gene><Tumor Suppression><Universities><Unscheduled DNA Synthesis><Xenograft><Xenograft procedure><Xenotransplantation><alzheimer model><astrocytic glia><bacteriophage P1 recombinase Cre><biological signal transduction><brain irradiation><brain radiation><cancer cell><cancer microenvironment><chemical library><chimeric antigen T cell 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><chronic lymphoid leukemia><chronic traumatic encephalopathy><clinical applicability><clinical application><cognitive dysfunction><cognitive loss><cranial radiation><degenerative diseases of motor and sensory neurons><degenerative neurological diseases><developmental><drug candidate><drug repositioning><drug repurposing><drug/agent><genome mutation><glioblastoma multiforme><hepatic damage><hepatic injury><high throughput screening><improved><in vivo><in vivo Model><inducible expression><inducible gene expression><knock-down><knockdown><knockin mice><lipid based nanoparticle><lipid nanoparticle><liver damage><liver injury><lung cancer><lung cancer cell><malignancy><metabolome><metabonome><migration><mouse model><murine model><mutant><naturally occurring product><necrocytosis><neoplasm/cancer><neural degeneration><neurodegeneration><neurodegenerative><neurodegenerative illness><neurological degeneration><neuronal degeneration><non-oncogenic><nononcogenic><novel><nutrition><p53 Antigen><p53 Genes><p53 Tumor Suppressor><pathway><protein p53><radiation treatment><repurposing agent><repurposing medication><senescence><senescence associated secretome><senescence associated secretory phenotype><senescent><siRNA><small molecule libraries><spongioblastoma multiforme><traumatic brain damage><treatment with radiation><tumor><tumor microenvironment><tumorigenesis><xeno-transplant><xeno-transplantation>