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Principal Investigator: MARK MERCOLA
Organization: STANFORD UNIVERSITY
Fiscal Year: 2024
Award: $597,054
Funding agency: National Heart Lung and Blood Institute
PROJECT SUMMARY
Cardiovascular complications of cancer therapy significantly contribute to the global burden of cardiovascular
diseases. Although remarkable progress has been made in understanding the genetic basis of doxorubicin-
induced cardiotoxicity (DIC), we cannot predict which patients will be affected by DIC or protect patients at risk
for suffering from DIC adequately. Here, we will use a novel multiplexing methodology of creating a "cell village"
by pooling multiple patients' induced pluripotent stem cell (iPSC) lines in a dish to map the genetic basis of inter-
individual differences in response to doxorubicin. In Aim 1, we will co-culture 100 iPSC lines in 10 distinct "cell
villages," where each "cell village" contains ten independent patient-specific iPSC lines. Next, we will differentiate
each "cell village" into iPSC-derived cardiomyocytes (iPSC-CMs). Finally, we will perform a single-cell multi-
omics sequencing analysis of the "cell villages" to understand the impact of genetic variability on cardiomyocyte
gene regulation and functions at baseline. In Aim 2, we will employ a single-cell multi-omic approach to uncover
and validate the role of response eQTL in DIC prediction. We will treat iPSC-CMs in each "cell village" with
doxorubicin at various doses. Next, we will perform single-cell multi-omics profiling to model the contribution of
genetics to variability in responses to doxorubicin treatment. In Aim 3, we will employ 3D engineered heart tissues
(EHTs) and CRISPR/cas9 genome-editing to comprehensively study the functional role of two candidate
doxorubicin response genes. All in all, the proposed experiments will serve as a proof-of- principle in using the
"cell village" model as a high throughput personalized drug screening platform.
Terms: <14-Hydroxydaunomycin><3-D><3-Dimensional><3D><ATAC><Adriamycine><Adverse effects><Affect><Aging><Alleles><Allelomorphs><Biological Function><Biological Process><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><Calcium><Cancer Treatment><Cardiac Muscle Cells><Cardiac Myocytes><Cardiac Toxicity><Cardiocyte><Cardiotoxic><Cardiotoxicity><Cardiovascular><Cardiovascular Body System><Cardiovascular Diseases><Cardiovascular Organ System><Cardiovascular system><Cas nuclease technology><Causality><Cell Body><Cell Line><Cell Survival><Cell Viability><CellLine><Cells><Chromosome Mapping><Circulation><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><Co-culture><Cocultivation><Coculture><Coculture Techniques><Coupled><DNA Damage><DNA Injury><Dose><Doxorubicin><Doxorubicina><Drug Screening><Drugs><Etiology><Exposure to><Gene Action Regulation><Gene Expression><Gene Expression Regulation><Gene Localization><Gene Mapping><Gene Mapping Genetics><Gene Regulation><Gene Regulation Process><Gene variant><Genes><Genetic><Genotype><Grant><Heart><Heart Muscle Cells><Heart Vascular><Heart myocyte><Hour><Hydroxyl Daunorubicin><Hydroxyldaunorubicin><Individual><Individual Differences><Investigators><Journals><Knowledge><LPTN><Linkage Mapping><Magazine><Malignant Neoplasm Therapy><Malignant Neoplasm Treatment><Maps><Medication><Methodology><Modeling><NR1B3><Nature><Patients><Pharmaceutical Preparations><Population Heterogeneity><Predisposition><Production><Progenitor Cells><Publishing><QTL><Quantitative Trait Loci><RARC><RARG><RARG gene><Research Personnel><Researchers><Risk><Role><SCM-1><SCM-1a><SCM1><SCYC1><Series><Single cell seq><Staining method><Stains><Strains Cell Lines><Susceptibility><Technology><Time><Total Human and Non-Human Gene Mapping><Toxic effect><Toxicities><Transcription Initiation Site><Transcription Start Site><XCL1><XCL1 gene><allele variant><allelic variant><anti-cancer therapy><cancer complication><cancer therapy><cancer-directed therapy><cardiac tissue engineering><cardiomyocyte><cardiovascular disorder><causation><circulatory system><cultured cell line><cytotoxic><differentiation protocol><disease causation><diverse populations><dosage><drug/agent><engineered heart tissue><entire genome><epigenome><experiment><experimental research><experimental study><experiments><full genome><gene function><genetic mapping><genetic variant><genome editing><genome scale><genome sequencing><genome-wide><genomewide><genomic editing><genomic variant><global gene expression><global transcription profile><heterogeneous population><hiPSC><human iPS><human iPSC><human induced pluripotent cell><human induced pluripotent stem cells><human inducible stem cells><iPS><iPS cell derived cardiomyocytes><iPSC><iPSC derived cardiomyocytes><iPSCs><induced human pluripotent stem cells><induced pluripotent cell><induced pluripotent stem cell><induced pluripotent stem cell derived cardiomyocytes><induced pluripotent stem cells derived from patients><induced pluripotent stem cells from patients><inducible pluripotent stem cell><inter-individual variability><inter-individual variation><interindividual variability><interindividual variation><multiomics><multiple omics><novel><panomics><patient derived human iPS><patient derived human iPSC><patient derived human induced pluripotent stem cell><patient derived iPS><patient derived iPSC><patient derived induced pluripotent cells><patient derived induced pluripotent stem cells><patient-derived pluripotent stem cells><personalized drugs><population diversity><precision drugs><response><scATAC sequencing><scATAC-seq><scRNA-seq><single cell ATAC-seq><single cell ATAC-sequencing><single cell Assay for Transposase Accessible Chromatin sequencing><single cell RNA-seq><single cell RNAseq><single cell analysis><single cell expression profiling><single cell next generation sequencing><single cell sequencing><single cell sequencing assay for transposase accessible chromatin><single cell transcriptomic profiling><single-cell Assay for Transposase-Accessible Chromatin with sequencing><single-cell RNA sequencing><single-cell assay for transposase-accessible chromatin using sequencing><single-cell assay for transposase-accessible chromatin-seq><social role><stem cells><three dimensional><transcriptome><whole genome>