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Principal Investigator: Cole A DeForest
Organization: UNIVERSITY OF WASHINGTON
Fiscal Year: 2024
Award: $428,361
Funding agency: National Cancer Institute
PROJECT SUMMARY
Despite innovations in diagnosis and treatment, colorectal cancer (CRC) remains one the most common causes
of cancer-related deaths in the United States. During tumorigenesis, the extracellular matrix (ECM) drastically
stiffens, resulting in a self-amplifying feedback loop, furthering cancer progression; in CRC, increased matrix
elasticity is correlated with more advanced stages and worse treatment outcomes. Cells sense tissue stiffness
via integrins, which relay biochemical signals down many pathways, including the Rho/ROCK pathway. The
Rho/ROCK pathway has been shown to be highly dysregulated in many cancers, but its role in CRC is still highly
debated: some studies have shown elevated levels in metastatic lesions, whereas others have correlated
inactivation of this pathway with metastasis. Herein, we propose to study the mechanical contributions to RhoA
activation and how this may lead to increased proliferation and signaling, with a particular emphasis on
comparing the mechanical environments of the colon and the liver—the most common metastatic site, which has
been shown to be significantly stiffer than the primary tumor tissue. We will develop a novel hydrogel biomaterial
that can stiffen around cells from 1kPa (primary tumor) to 6kPa (liver metastatic tumor) upon exposure to 365nm
light over the course of the week, by employing photomediated oxime ligation, a chemistry we have pioneered
in our previous work for making photostiffening hydrogels as well as immobilizing full-length proteins in natural
and synthetic materials. To encode for subsequent cell release for downstream assays, we will develop a suite
of enzymatically sensitive peptide crosslinkers to enable bioorthogonal, user-defined hydrogel degradation. In
Aim 2, we will examine RhoA activation in primary patient-derived CRC tumor organoids as a function of
progressive stiffening of the matrix, and compare to levels of RhoA activation in metastatic cancer organoid lines
in stiff gels, via pull-down assays, Western blotting, and a lentivirally transduced biosensor construct. In Aim 3,
we will investigate the mechanical memory of RhoA activation in metastatic tumor organoid lines and whether is
reversable upon systematic softening of the matrix back to baseline, primary colon stiffnesses. We expect results
from these studies will result in novel insights into the role of matrix mechanics in CRC progression and
potentially open the door for biophysical routes of tumor resolution and treatment.
Terms: <1,2-Ethanediol><2-Hydroxyethanol><3-D><3-Dimensional><3D><Abscission><Actins><Assay><Atomic Force Microscopy><Attenuated><Back><Binding><Bioassay><Biochemical><Biocompatible Materials><Biological Assay><Biomaterials><Biophysics><Biosensor><Body Tissues><Cancer Cause><Cancer Etiology><Cancers><Cell Body><Cell Communication and Signaling><Cell Signaling><Cell division><Cell-Extracellular Matrix><Cells><Cessation of life><Chemistry><Closure by Ligation><Colon><Colon Cancer><Colon Carcinoma><Colorectal Cancer><Crosslinker><Death><Development><Diagnosis><Dihydroxyethanes><Disease Progression><Disputes><Disseminated Malignant Neoplasm><Dorsum><ECM><Elasticity><Encapsulated><Environment><Enzyme Gene><Enzymes><Ethanediols><Ethylene Glycols><Event><Excision><Exposure to><Extirpation><Extracellular Matrix><Feedback><Force Microscopy><Future><Gel><Generalized Growth><Goals><Growth><Hepatic Neoplasm Secondary><Hepatic metastasis><Hydrogels><Hydroxyimino Compounds><Immunoblotting><In Situ><Integrin Binding><Integrins><Integrins Extracellular Matrix><Intracellular Communication and Signaling><Lead><Length><Lesion><Ligands><Ligation><Light><Liver><Liver secondaries><Liver secondary cancer><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Measures><Mechanics><Mediating><Mediator><Memory><Metastasis><Metastasize><Metastatic Cancer><Metastatic Lesion><Metastatic Malignant Neoplasm><Metastatic Mass><Metastatic Neoplasm><Metastatic Neoplasm to the Liver><Metastatic Tumor><Metastatic Tumor to the Liver><Metastatic malignant neoplasm to liver><Modeling><Molecular Interaction><Monitor><Monoethylene Glycol><Morphology><Movement><Myosin Light Chains><Neoplasm Metastasis><Oncogenesis><Operative Procedures><Operative Surgical Procedures><Organoids><Outcome><Oximes><Pathogenesis><Pathway interactions><Patients><Pb element><Peptides><Phenotype><Phosphorylation><Photolyses><Photoradiation><Physiologic><Physiological><Primary Lesion><Primary Neoplasm><Primary Tumor><Proliferating><Protein Phosphorylation><Proteins><Psychological reinforcement><RTKN><Recovery><Reinforcement><Removal><Reporting><Resolution><Role><Route><Sampling><Scaffolding Protein><Scanning Force Microscopy><Secondary Neoplasm><Secondary Tumor><Signal Transduction><Signal Transduction Systems><Signaling><Site><Spectroscopy><Spectrum Analyses><Spectrum Analysis><Stimulus><Surgical><Surgical Interventions><Surgical Procedure><Surgical Removal><Survival Rate><Suspension substance><Suspensions><System><Time><Tissue Growth><Tissues><Treatment outcome><Tumor Cell><Tumor Tissue><United States><Visualization><Western Blotting><Western Immunoblotting><Work><antifibrotic agent><antifibrotic medication><antifibrotic therapy><antifibrotic treatment><attenuate><attenuates><biocompatibility><biological material><biological sensor><biological signal transduction><biomaterial compatibility><biophysical foundation><biophysical principles><biophysical sciences><body movement><cancer cell><cancer in the colon><cancer metastasis><cancer progression><colon cancer patients><colorectal cancer patients><colorectal cancer progression><design><designing><developmental><dosage><ethylene glycol><extracellular><heavy metal Pb><heavy metal lead><hepatic body system><hepatic organ system><in vivo><in vivo Model><innovate><innovation><innovative><insight><integrin bound><lentiviral-transduced><lentivirally transduced><lentivirus transduced><light microscopy><liver metastases><malignancy><malignant liver neoplasm, specified as secondary><mechanic><mechanical><metastasis in the liver><metastasis to the liver><metastasize to the liver><metastatic cancer to liver><metastatic colo-rectal><metastatic colo-rectal cancer><metastatic colo-rectal carcinoma><metastatic colon cancer><metastatic colorectal><metastatic colorectal cancer><metastatic colorectal carcinoma><metastatic liver><metastatic liver neoplasm><neoplasm progression><neoplasm/cancer><neoplastic cell><neoplastic progression><novel><ontogeny><pathway><peptide aminoacid sequence><peptide sequence><pharmacologic><photolysis><protein aminoacid sequence><protein blotting><recruit><resection><resolutions><response><rho><rhotekin><scaffold><scaffolding><secondary liver malignancy><secondary malignant liver neoplasm><social role><sortase><srtA gene product><sugar><surgery><three dimensional><tumor><tumor cell metastasis><tumor progression><tumorigenesis>