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Principal Investigator: Shira Yomtoubian
Organization: SALK INSTITUTE FOR BIOLOGICAL STUDIES
Fiscal Year: 2024
Award: $76,756
Funding agency: National Cancer Institute
Project Summary
Lung cancer is the leading cause of cancer-related mortality worldwide. Lung tumors driven by mutant KRAS
are among the most aggressive and refractory to treatment, due in part by KRAS-driven metabolic
reprogramming. Efforts to better understand the pathways regulating metabolic adaptations in oncogenic KRAS-
driven tumors will provide insight into lung cancer progression and identify vulnerabilities that could be
therapeutically targeted to improve patient survival. The Shaw lab recently showed the requirement of AMP-
activated protein kinase (AMPK) to promote the growth of oncogenic KRAS-driven non-small-cell lung cancer
(NSCLC). AMPK is a master regulator of cellular and organismal metabolism that acts as a sensor of cellular
energy by altering metabolism when energy levels are low. While the Shaw lab and others have demonstrated
that AMPK signaling provides cancer cells with flexibility to adapt to metabolic stresses, the epigenetic
mechanisms by which AMPK promotes metabolic alterations and lung tumor growth remain poorly understood.
Preliminary studies identified a de novo DNA methyltransferase as a novel substrate of AMPK. DNA methylation
is involved in many normal cellular processes and is abnormally distributed in cancer cells, contributing to some
of their aggressive characteristics. This proposal addresses the consequences of AMPK-dependent regulation
of the de novo DNA methyltransferase on DNA methylation, metabolic programs, and lung tumor growth. First,
this work aims to define the methylation profile controlled by the AMPK-dependent phosphorylated form of the
de novo DNA methyltransferase using whole-genome bisulfite sequencing, CUT&TAG, and RNA sequencing
assays. Tumorigenicity and Seahorse real-time cell metabolic analyses will determine whether this regulation
disrupts DNA methylation patterns in a manner that generates tumor-promoting epigenetic lesions and metabolic
alterations. Additionally, the generation of autochthonous KRAS-driven NSCLC mouse lines expressing
constitutive knock-in of the de novo DNA methyltransferase with a serine-to-alanine mutation at the putative
phosphor-acceptor-serine will enable testing whether regulation of the de novo DNA methyltransferase impacts
tumor initiation, growth, and metastasis. This work addresses a fundamental relationship between two hallmarks
of cancer and if successful would lead to the mechanistic connection between metabolic stresses tumor cells
face and how they may trigger sustained DNA methylation changes.
Terms: <(hydroxymethylglutaryl-CoA reductase (NADPH)) kinase><5'-AMP-activated protein kinase><AMP-activated kinase><AMP-activated protein kinase><AMPK enzyme><Address><Alanine><Antiphosphopeptide-Specific Antibodies><Assay><BS-seq><Binding><Bio-Informatics><Bioassay><Bioinformatics><Biological Assay><Bisulfite-based sequencing><Cancer Cause><Cancer Cell Growth><Cancer Etiology><Cancer Patient><Cancer cell line><Cancers><Cell Body><Cell Communication and Signaling><Cell Function><Cell Growth in Number><Cell Multiplication><Cell Physiology><Cell Process><Cell Proliferation><Cell Respiration><Cell Signaling><Cells><Cellular Function><Cellular Physiology><Cellular Process><Cellular Proliferation><Cellular Respiration><Characteristics><Common Rat Strains><Consensus><DNA Methylation><DNA Methylation Regulation><DNA Methyltransferase><DNA Modification Methylases><DNA Modification Methyltransferases><DNA-Methyltransferases><DNMT3a><Dnmt><EC 2.1.1><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Exhibits><Face><GeneHomolog><Generalized Growth><Generations><Genetic><Genetic Alteration><Genetic Change><Genetic defect><Genus Hippocampus><Growth><HMG CoA reductase (NADPH) kinase><HMG CoA reductase kinase><HMG coenzyme A reductase (NADPH) kinase><Homolog><Homologous Gene><Homologue><Impairment><In Vitro><Intermediary Metabolism><Intracellular Communication and Signaling><Invaded><KRAS(G12D)><KRASG12D><Knock-in><Knock-out><Knockout><L-Serine><Lesion><Lung Neoplasms><Lung Tumor><Malignant Cell><Malignant Neoplasms><Malignant Soft Tissue Neoplasm><Malignant Tumor><Malignant Tumor of the Lung><Malignant neoplasm of lung><Mediating><Metabolic><Metabolic Processes><Metabolic stress><Metabolism><Metastasis><Metastasize><Metastatic Lesion><Metastatic Mass><Metastatic Neoplasm><Metastatic Tumor><Methylation><Methyltransferase><Mice><Mice Mammals><Mitochondria><Modeling><Modification Methylases><Molecular Interaction><Murine><Mus><Mutation><NSCLC><NSCLC - Non-Small Cell Lung Cancer><Neoplasm Metastasis><Non-Small Cell Lung Cancer><Non-Small-Cell Lung Carcinoma><Oncogenic><Pathway interactions><Patients><Phenotype><Phospho-Specific Antibodies><Phosphopeptide-Specific Antibodies><Phosphorylation><Phosphorylation State-Specific Antibodies><Phosphospecific Antibody><Proliferating><Protein Phosphorylation><Proteins><Pulmonary Cancer><Pulmonary Neoplasms><Pulmonary malignant Neoplasm><RNA Seq><RNA sequencing><RNAseq><Rat><Rats Mammals><Rattus><Refractory><Regulation><Reporting><Role><Sarcoma><Seahorse><Secondary Neoplasm><Secondary Tumor><Serine><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Site-Specific DNA-methyltransferase><Subcellular Process><Testing><Time><Tissue Growth><Tumor Burden><Tumor Cell><Tumor Load><Tumor Promotion><Tumor Suppressor Proteins><Tumorigenicity><Validation><Viral Oncogene><Work><aerobic metabolism><aerobic respiration><biological signal transduction><bisulfite sequencing><bisulfite-seq><cancer cell><cancer metastasis><cancer progression><colonization associated with lung><colonization in the lung><colonization within the lung><entire genome><epigenetically><epigenome><faces><facial><flexibility><flexible><full genome><genome mutation><hDNA methyltransferase 3a><hydroxymethylglutaryl-CoA-reductase kinase><improved><in vivo><insight><knockin><lung cancer><lung cancer cell><lung colonization><malignancy><malignant soft tissue tumor><methylase><methylation pattern><migration><mitochondrial><mortality><mouse model><murine model><mutant><neoplasm progression><neoplasm/cancer><neoplastic cell><neoplastic progression><novel><ontogeny><oxidative metabolism><pathway><programs><pulmonary colonization><sensor><social role><targeted drug therapy><targeted drug treatments><targeted therapeutic><targeted therapeutic agents><targeted therapy><targeted treatment><therapeutic target><therapeutically effective><transcriptome sequencing><transcriptomic sequencing><transmethylase><tumor><tumor cell metastasis><tumor growth><tumor initiation><tumor progression><tumor suppressor><tumorigenic><validations><whole genome>