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Principal Investigator: Yun Wei
Organization: MASSACHUSETTS GENERAL HOSPITAL
Fiscal Year: 2024
Award: $178,200
Funding agency: National Cancer Institute
PROJECT SUMMARY / ABSTRACT
Rhabdomyosarcomas (RMS) are the most common childhood soft-tissue sarcomas affecting hundreds of
patients in the United States annually. Current standard treatments for rhabdomyosarcoma (RMS) patients
include chemotherapy, surgery, and /or radiation. However, even with these combinations of therapeis,
significant subsets of patients suffer tumor recurrence, relapse, and metastasis, associated with extremely worse
prognosis and dismal 5-year survival rate. This remains the major hurdle to improve the patient outcomes with
rhabdomyosarcomas. To better understand the mechanisms such as tumor-propagating cells, critical molecular
regulators that drives therapy resistance and tumor-relapse in RMS, researchers has employed RMS cell lines,
transgenic animal models, and xenograft studies to study the potential tumor-propagating cells (TPCs) for RMS.
Yet, little is known about the tumor heterogeneity and cancer cell evolution dynamics in RMS. To dissect the
inter-tumoral and intra-tumoral heterogeneity, I have used the single-cell transcriptomics to profile patient-derived
samples of RMS. I uncovered distinct cell states in RMS tumors, including proliferation and a mesenchymal-like
subpopulations that have higher TPC potential, whereas the differentiated muscle subpopulation that barely
transits towards other cell states. With this knowledge, and the innovation of barcode tracing techniques, I
propose to dissect molecular mechanisms that contribute to cell state transitions, and concurrently assess the
cell phenotypes changes along with its transcripts, proteins, and epigenetics alterations. One class of important
and challenging molecules in regulating cancer stemness, evolution post therapies is chromatin regulators, which
requires deep sequencing in limited cell line models. The technical innovation of single-cell multiomics, including
single-cell RNA, single-cell ATAC, single-cell CUT&Tag, and cell lineage barcode tracing largely decrease the
cost and time needed to profile cancer cell evolution along with epigenetic modifications at single-cell levels.
With effective collaboration with computational biologists, I hypothesize that EZH2 and its catalytic product
H3K27me3 lock RMS cells in the proliferative cell state and inhibit their transition into other differentiated states.
To test this hypothesis, I will first assess the role of EZH2 in regulating cell state transitions with barcode tracing
and functional stem assays in the context of EZH2 knockdown (Aim 1). Independently, I will also profile the direct
targets of EZH2 and histone H3 lysine 27 trimethylation by performing single-cell CUT&Tag, and interrogate
mechanism that controls cell state transition (Aim 2). In addition, I will also assess the EZH2 inhibitors in
collaboration with chemotherapy and radiation utilizing the unique immune-compromised zebrafish models along
with cell line and mouse xenograft studies (Aim 3). The goals of the proposed research are to investigate
chromatin regulators in rhabdomyosarcoma samples while also acknowledging the tumor-heterogeneity and cell
plasticity in cell state transitions. By achieving these aims, I will illustrate a comprehensive mechanism as to how
RMS tumors evolve and how chromatin regulators play critical roles in controlling this process.
Terms: <ATAC><Actinomycin A IV><Actinomycin C1><Actinomycin D><Actinomycin I1><Actinomycin IV><Actinomycin X 1><Affect><After Care><After-Treatment><Aftercare><Animal Model><Animal Models and Related Studies><Assay><Bar Codes><Basal Transcription Factor><Basal transcription factor genes><Binding><Bioassay><Biological Assay><Brachydanio rerio><CTX><CYCLO-cell><Cancers><Carloxan><Catalytic Core><Catalytic Domain><Catalytic Region><Catalytic Site><Catalytic Subunit><Cell Body><Cell Cycle><Cell Differentiation><Cell Differentiation process><Cell Division Cycle><Cell Growth in Number><Cell Line><Cell Lineage><Cell Multiplication><Cell Proliferation><CellLine><Cells><Cellular Proliferation><Chemotherapy and Radiation><Chemotherapy and/or radiation><Childhood Neoplasm><Childhood Soft Tissue Sarcoma><Childhood Tumor><Chromatin><Ciclofosfamida><Ciclofosfamide><Cicloxal><Clafen><Claphene><Clinic><Collaborations><Complex><Cosmegen><Cycloblastin><Cycloblastine><Cyclophospham><Cyclophosphamide><Cyclophosphamidum><Cyclophosphan><Cyclophosphane><Cyclophosphanum><Cyclostin><Cyclostine><Cytophosphan><Cytophosphane><Cytoxan><Dactinomycin><Dactinomycine><Danio rerio><Drosophila Homolog of NOTCH 3><Drugs><ENX-1><EZH1><EZH2><EZH2 gene><Embryonic Muscle Cells><Endoxan><Endoxana><Enduxan><Enhancer of Zeste 2 Polycomb Repressive Complex 2 Subunit><Epigenetic><Epigenetic Change><Epigenetic Mechanism><Epigenetic Process><Evolution><Fosfaseron><Frequencies><Gene Down-Regulation><Gene Expression><Gene Transcription><General Transcription Factor Gene><General Transcription Factors><Genetic Transcription><Genoxal><Genuxal><Goals><Heterogeneity><Heterograft><Heterologous Transplantation><Histone H3><Histones><Image><Immune><Immunes><Impairment><Inter-tumoral heterogeneity><Intratumoral heterogeneity><Investigators><KMT6><KMT6A><Knowledge><L-Lysine><LPTN><Ledoxina><Leurocristine><Lyovac cosmegen><Lysine><MEF-2C><MEF2C protein><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Medication><Meractinomycin><Mesenchymal><Messenger RNA><Metastasis><Metastasize><Metastatic Lesion><Metastatic Mass><Metastatic Neoplasm><Metastatic Tumor><Methylation><Mice><Mice Mammals><Mitoxan><Modeling><Modification><Molecular><Molecular Interaction><Murine><Mus><Muscle><Muscle Cells><Muscle Fibers><Muscle Tissue><Mutate><Myoblasts><Myocytes><Myotubes><NOTCH3><NOTCH3 gene><Nature><Neoplasm Metastasis><Neosar><Non-Polyadenylated RNA><Operative Procedures><Operative Surgical Procedures><PDX model><Pathway interactions><Patient derived xenograft><Patient outcome><Patient-Centered Outcomes><Patient-Focused Outcomes><Patients><Pediatric Neoplasm><Pediatric Soft Tissue Sarcoma><Pediatric Tumor><Pharmaceutical Preparations><Phenotype><Play><Polycomb><Precursor Muscle Cells><Process><Procytox><Prognosis><Proliferating><Proteins><RNA><RNA Expression><RNA Gene Products><Radiation><Recurrence><Recurrent><Recurrent Neoplasm><Recurrent tumor><Relapse><Repressor Proteins><Research><Research Personnel><Researchers><Resistance><Resolution><Rhabdomyocyte><Rhabdomyosarcoma><Ribonucleic Acid><Role><SCM-1><SCM-1a><SCM1><SCYC1><Sampling><Secondary Neoplasm><Secondary Tumor><Sendoxan><Skeletal Fiber><Skeletal Muscle Cell><Skeletal Muscle Fiber><Skeletal Myocytes><Stem Cell Assay><Stem Cell like><Strains Cell Lines><Stress><Surgical><Surgical Interventions><Surgical Procedure><Survival Rate><Syklofosfamid><Tail><Techniques><Testing><Time><Transcript><Transcription><Transcription Activator><Transcription Coactivator><Transcription Factor Coactivator><Transcription Factor Proto-Oncogene><Transcription Repression><Transcription factor genes><Transcriptional Activator><Transcriptional Activator/Coactivator><Transcriptional Coactivator><Transcriptional Repression><Transgenic Animals><Tumor Cell><Undifferentiated><United States><Vincristine><Vincrystine><Work><XCL1><XCL1 gene><Xenograft><Xenograft procedure><Xenotransplantation><Zebra Danio><Zebra Fish><Zebrafish><Zytoxan><barcode><cancer cell><cancer metastasis><cell killing><cellular differentiation><chemo/radiation therapy><chemotherapy><chemotherapy and radiotherapy><childhood sarcoma><cost><cultured cell line><deep sequencing><drug detection><drug testing><drug/agent><dynamical evolution><epigenetically><gene repression><heterogeneity in tumors><histone methylation><imaging><improved><in vivo><inhibitor><innovate><innovation><innovative><intertumoral heterogeneity><intra-tumoral heterogeneity><intratumor heterogeneity><irradiation><knock-down><knockdown><mRNA><malignancy><model of animal><mouse model><multiomics><multiple omics><murine model><muscular><myocyte enhancer factor 2C><myocyte-specific enhancer-binding-factor 2C><neoplasm recurrence><neoplasm/cancer><neoplastic cell><new drug target><new drug treatments><new druggable target><new drugs><new pharmacological therapeutic><new pharmacotherapy target><new therapeutic target><new therapeutics><new therapy><new therapy target><next generation therapeutics><novel><novel drug target><novel drug treatments><novel druggable target><novel drugs><novel pharmaco-therapeutic><novel pharmacological therapeutic><novel pharmacotherapy target><novel therapeutic target><novel therapeutics><novel therapy><novel therapy target><oxovincaleukoblastine><panomics><pathway><patient derived xenograft model><patient oriented outcomes><patient profile><patient subclass><patient subcluster><patient subgroups><patient subpopulations><patient subsets><patient subtypes><pediatric sarcoma><pharmacologic><post treatment><pre-clinical><pre-clinical efficacy><preclinical><preclinical efficacy><profiles in patients><progenitor><progenitor cell assay><programs><radiation or chemotherapy><repressor complex><resistance to therapy><resistant><resistant to therapy><resolutions><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 expression profiling><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><standard care><standard treatment><stem><stem cell characteristics><stemness><surgery><synergism><therapeutic resistance><therapy resistant><transcription co-activator><transcription factor><transcriptional co-activator><transcriptomics><treatment resistance><tumor><tumor cell metastasis><tumor growth><tumor heterogeneity><tumors in children><xeno-transplant><xeno-transplantation>