Document text
Principal Investigator: Roberto Hugo Barbier
Organization: DUKE UNIVERSITY
Fiscal Year: 2024
Award: $41,798
Funding agency: National Cancer Institute
ABSTRACT
Prognosis for patients with small cell lung cancer (SCLC) remains poor due to high rates of metastatic disease,
vast intra-tumoral heterogeneity, limited therapeutic options, and rapid development of therapeutic resistance.
Our lab has recently shown that Abelson (ABL) family kinase inhibitors are effective in the treatment of SCLC
metastases in vivo. Targeted combination therapies can minimize cytotoxic adverse events and impair the
development of resistance. Thus, we performed a whole-genome CRISPR/Cas9 loss-of-function sensitization
screen to identify synergizing combination therapy targets with the ABL allosteric inhibitor ABL001. Consistent
with prior observations that ABL kinase inhibition dysregulates mitochondrial and metabolic function in lung
adenocarcinoma, numerous top sensitizers to ABL kinase inhibition revealed by the screen are involved in the
processing and clearance of metabolic products. Preliminary validation of the screen has shown that inhibition
of organic anion MCT transporters synergizes with ABL001 to cause cell death as a consequence of a
mitochondrial dysregulation phenotype observed in lung cancer cells following treatment with allosteric ABL
kinase inhibitors. Furthermore, targeted metabolomic sequencing revealed that following combination treatment
with ABL and MCT inhibitors, the levels of numerous metabolite species become significantly dysregulated.
These changes are consistent with the stalling of glutaminolysis and are most strikingly characterized by a
significant decrease in glutathione, an important mediator of metabolic stress responses. My hypothesis is that
the metabolic stress induced by inhibition of the ABL kinases when paired with MCT inhibition, results in the
stalling of glutaminolysis and the subsequent loss of glutathione, thereby promoting ferroptosis, and contributing
to the synergistic cell death phenotype observed. The aims of this proposal are 1) to determine how the ABL
kinases mediate homeostatic metabolic function and define the mechanism of metabolic dysregulation following
ABL kinase inhibition, 2) to determine the mechanism of dysregulation of glutaminolysis following combined ABL
kinase and MCT inhibition, and 3) assess the extent to which this combined therapy contributes to cell death via
ferroptosis. Approaches to address these objectives include metabolic profiling via stable isotope tracing of
glucose and glutamine, and Seahorse assays to measure oxidative and glycolytic energy metabolism. We will
also assess the extent to which this combination treatment induces ferroptotic cell death by in vitro drug-
treatment and isogenic knockout experiments. Subsequently we will perform in vivo drug treatment experiments
to assess the efficacy of combined ABL and MCT inhibition in the treatment of SCLC in mouse models. This
investigation into the role of the ABL kinases in metabolic dynamics will explore a novel mechanism of regulation
of mitochondrial homeostasis by the ABL kinases, as well as evaluate a potential treatment modality for patients
with SCLC.
Terms: <Active Oxygen><Address><Adverse Experience><Adverse event><Anions><Assay><Athymic Mice><Athymic Nude Mouse><Autoregulation><Bioassay><Biochemical Pathway><Biological Assay><CD147 antigen><CRISPR approach><CRISPR based approach><CRISPR method><CRISPR methodology><CRISPR technique><CRISPR technology><CRISPR tools><CRISPR-CAS-9><CRISPR-based method><CRISPR-based technique><CRISPR-based technology><CRISPR-based tool><CRISPR/CAS approach><CRISPR/Cas method><CRISPR/Cas technology><CRISPR/Cas9><CRISPR/Cas9 technology><Cancer cell line><Cas nuclease technology><Cell Body><Cell Death><Cell Death Induction><Cells><Chaperone><Clustered Regularly Interspaced Short Palindromic Repeats approach><Clustered Regularly Interspaced Short Palindromic Repeats method><Clustered Regularly Interspaced Short Palindromic Repeats methodology><Clustered Regularly Interspaced Short Palindromic Repeats technique><Clustered Regularly Interspaced Short Palindromic Repeats technology><Collaborations><Combined Modality Therapy><D-Glucose><Data><Dependence><Dextrose><Disease><Disorder><Disseminated Malignant Neoplasm><Drug Therapy><Energy Expenditure><Energy Metabolism><Energy Metabolism - Reference Pathway><Energy Metabolism Pathway><FDA approved><Family><Flow Cytofluorometries><Flow Cytofluorometry><Flow Cytometry><Flow Microfluorimetry><Flow Microfluorometry><Gene Down-Regulation><Gene Transcription><Genetic><Genetic Transcription><Genus Hippocampus><Gln><Glucose><Glutamine><Glutathione><Goals><HT7 antigen><Heterogeneity><Homeostasis><Human><Immunocompetent><Impairment><Implant><In Vitro><Injections><Intratumoral heterogeneity><Investigation><Kinases><Knock-out><Knockout><Knowledge><L-Glutamine><Lipid Peroxidation><Lung Adenocarcinoma><Malignant Tumor of the Lung><Malignant neoplasm of lung><Measures><Mediating><Mediator><Metabolic><Metabolic Networks><Metabolic Pathway><Metabolic stress><Metastasis><Metastasize><Metastatic Cancer><Metastatic Lesion><Metastatic Malignant Neoplasm><Metastatic Mass><Metastatic Neoplasm><Metastatic Tumor><Mitochondria><Modality><Modern Man><Molecular Chaperones><Multimodal Therapy><Multimodal Treatment><Nature><Neoplasm Metastasis><Nude Mice><Oat cell carcinoma><Oxidative Phosphorylation><Oxidative Phosphorylation Pathway><Oxidative Stress><Oxidative Stress Induction><Oxygen Radicals><Patients><Pharmacotherapy><Phenotype><Phosphotransferase Gene><Phosphotransferases><Physiological Homeostasis><Pro-Oxidants><Pulmonary Cancer><Pulmonary malignant Neoplasm><Q Levoglutamide><Q. Levoglutamide><RNA Expression><Radiation><Reactive Oxygen Species><Regulation><Research><Resistance><Resistance development><Resistant development><Role><Seahorse><Secondary Neoplasm><Secondary Tumor><Small Cell Lung Cancer><Stable Isotope Labeling><System><Therapeutic><Transcription><Transcription Repression><Transcriptional Repression><Transphosphorylases><Validation><Work><assess effectiveness><basigin><biological adaptation to stress><cancer metastasis><cancer sub-types><cancer subtypes><combat><combination therapy><combined modality treatment><combined treatment><cytotoxic><deprivation><determine effectiveness><determine efficacy><developing resistance><drug treatment><effectiveness assessment><effectiveness evaluation><efficacy analysis><efficacy assessment><efficacy determination><efficacy evaluation><efficacy examination><efficacy testing><entire genome><evaluate effectiveness><evaluate efficacy><examine effectiveness><examine efficacy><experiment><experimental research><experimental study><experiments><flow cytophotometry><full genome><gamma-L-Glu-L-Cys-Gly><gamma-L-Glutamyl-L-Cysteinylglycine><gene repression><genome scale><genome-wide><genomewide><heterogeneity in tumors><immune competent><improved><in vivo><inhibitor><intra-tumoral heterogeneity><intratumor heterogeneity><kinase inhibitor><knock-down><knockdown><loss of function><lung cancer><lung cancer cell><lung oat cell carcinoma><lung small cell neuroendocrine carcinoma><metabolic profile><metabolism measurement><metabolomics><metabonomics><mitochondrial><mouse model><multi-modal therapy><multi-modal treatment><murine model><necrocytosis><new drug treatments><new drugs><new pharmacological therapeutic><new therapeutics><new therapy><next generation therapeutics><novel><novel drug treatments><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel therapeutics><novel therapy><oat cell cancer><overexpress><overexpression><patient prognosis><pharmacologic><pyr translocator><pyruvate carrier><pyruvate transport protein><pyruvate transporter><reaction; crisis><resistance to therapy><resistant><resistant to therapy><shRNA><short hairpin RNA><small cell lung carcinoma><small cell undifferentiated carcinoma><small hairpin RNA><social role><stable isotope><stress response><stress; reaction><synergism><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic agent development><therapeutic development><therapeutic resistance><therapy resistant><treatment resistance><tumor><tumor cell metastasis><tumor heterogeneity><validations><whole genome>