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Principal Investigator: Francis Young-In Lee
Organization: YALE UNIVERSITY
Fiscal Year: 2024
Award: $507,749
Funding agency: National Institute of Arthritis and Musculoskeletal and Skin Diseases
Project Summary:This A1 proposal focuses on impaired pathological fracture healing in the presence of breast
cancer cells, not on the entire complex sequence of cancer spread from the breast to bone. Metastatic cancer
cells can settle in anywhere in the bone including bone marrow, cortical bone, or outside of the bone. Cancer
cells are spilled in and out of bone at the time of fracture. Pathological fracture calluses are deficient and very
often fail to ossify despite adjuvant therapies such radiation therapy and anti-resorptive agents including
Rank:Fc antibody (denosumab) or bisphosphonates (zoledronate). Failed fracture healing is not solely a result
of bone resorption. There is no mechanistic understanding as to why fractures do not heal well in the presence
of certain types of breast cancer cells. Unraveling the mechanisms underlying impaired fracture healing will
enable patients to benefit from future scientific endeavors employing mechanism-based treatments. If there is a
way of addressing impaired fracture repair while also inhibiting cancer growth in bone, the clinical care for
pathological fractures will be immensely impacted. We conducted whole transcriptome bulk RNAsequencing of
several different types of breast cancer cells that are grown in breast vs. bone. We observed that breast cancer
cells that inhibit fracture repair are pro-inflammatory with heightened MEK1-pERK1/2-cytokine-
hyperinflammation signaling in the bone microenvironment. Temporal expression of pro-Inflammatory cytokines
and bone-acting proteins such as TNF, interleukins, and sclerostin are completely deranged and prolonged
following femur fractures in the presence of breast cancer cells such as MDA231, HC1806, and 4T1 However,
there is a critical knowledge gap as to how these different breast cancer cell types affect chondro-progenitors,
osteoblasts, osteoclasts, and osteocytes within the intact fracture callus architecture as suggested by the
Reviewer. Two notable changes in this A1 proposal include the refinement of the animal models and a new set
of compelling spatial transcriptomic biology data incorporating the Reviewers’ critiques and suggestions. We
posit a central hypothesis that highly inflammatory pERK1/2-high breast cancer cells inhibit the normal fracture
callus formation and maturation by causing prolonged hyper-inflammation and subsequent derangements of
callus spatial transcriptomics. We propose 3 Specific Aims. Aim 1. To define temporal and spatial
transcriptomic changes of inflammatory, osteogenic, and chondrogenic lineage cells in structurally intact
fracture callus in vivo. Aim 2. To rescue impaired osteogenesis of osteoprogenitors, chondroprogenitors, and
periosteal cells in the presence breast cancer cells by inhibition of pERK1/2-induced inflammation ex vivo. Aim
3. In Vivo Pre-clinical Therapeutic Translation: To establish a mechanism-based rescue of impaired
pathological fracture healing by targeting specific pro-inflammatory kinases in vivo. We expect to establish a
novel therapeutic paradigm for devastating patients suffering from cancer and pathological fractures.
Terms: <(TNF)-α><3-D><3-Dimensional><3D><4T1><Address><Adjuvant Therapy><Affect><Animal Model><Animal Models and Related Studies><Anti-Cancer Agents><Antibodies><Antimorphic mutation><Antineoplastic Agents><Antineoplastic Drugs><Antineoplastics><Architecture><Biology><Biomechanics><Bisphosphonates><Blinded><Bone Formation><Bone Marrow><Bone Marrow Reticuloendothelial System><Bone Resorption><Bone callus><Bony Callus><Breast><Breast Cancer><Breast Cancer Cell><Cachectin><Callus><Cancer Drug><Cancer Patient><Cancers><Cell Body><Cell Communication and Signaling><Cell Lineage><Cell Signaling><Cells><Co-culture><Cocultivation><Coculture><Coculture Techniques><Complex><Critiques><Data><Differentiated Gene><Diffusion><Disseminated Malignant Neoplasm><Dominant Negative><Dominant-Negative Mutant><Dominant-Negative Mutation><Drug Therapy><ERK 1><ERK1><ERK1 Kinase><Engineering / Architecture><Extracellular Signal-Regulated Kinase 1><Femoral Fractures><Femur><Fracture><Fracture Healing><Future><Gene Expression><Generalized Growth><Genetic Suppression><Growth><Hematoma><Histology><Immunoblotting><Immunohistochemistry><Immunohistochemistry Cell/Tissue><Immunohistochemistry Staining Method><Impairment><Inflammation><Inflammatory><Interleukins><Intracellular Communication and Signaling><Kinases><Knowledge><MAP Kinase 3><MAP Kinase Kinase 1><MAP2K1><MAP2K1 gene><MAPK/ERK Kinase 1><MAPK3><MAPK3 Mitogen-Activated Protein Kinase><MAPK3 gene><MAPKK1><MEK-1><MEK1><MKK1><Macrophage-Derived TNF><Malignant Breast Neoplasm><Malignant Cell><Malignant Neoplasms><Malignant Tumor><Metastatic Cancer><Metastatic Malignant Neoplasm><Metastatic breast cancer><Mice><Mice Mammals><Mitogen-Activated Protein Kinase 3><Mitogen-Activated Protein Kinase 3 Gene><Mitogen-Activated Protein Kinase Kinase-1><Molecular Target><Monocyte-Derived TNF><Murine><Mus><NDC-Zoledronate><Neoplastic Disease Chemotherapeutic Agents><Non-Polyadenylated RNA><Osteoblasts><Osteoclastic Bone Loss><Osteoclasts><Osteocytes><Osteogenesis><P44ERK1><PRKMK1><PSTkinase p44mpk><Pathologic Fracture><Pathological fracture><Patients><Periosteal Cell><Pharmacotherapy><Phosphorylation><Phosphotransferase Gene><Phosphotransferases><Physiologic Ossification><Physiological Ossification><Protein Phosphorylation><Proteins><RNA><RNA Gene Products><Radiation therapy><Radiotherapeutics><Radiotherapy><Ribonucleic Acid><Sampling><Scientist><Secondary to><Signal Transduction><Signal Transduction Systems><Signaling><Site><Spontaneous Fractures><Staining method><Stains><Suggestion><TNF><TNF A><TNF Alpha><TNF gene><TNF-α><TNFA><TNFα><Testing><Therapeutic><Therapeutic Trials><Time><Tissue Growth><Transfection><Transphosphorylases><Tumor Necrosis Factor><Tumor Necrosis Factor-alpha><Tumor-Specific Treatment Agents><Western Blotting><Western Immunoblotting><Woman><X-ray microtomography><Xray microtomography><Zoledronate><Zometa><adjuvant treatment><anti-cancer drug><biological signal transduction><biomechanical><biomechanical analyses><biomechanical analysis><biomechanical assessment><biomechanical characterization><biomechanical evaluation><biomechanical measurement><biomechanical profiling><biomechanical test><biphosphonate><bisphosphonate><bone><bone fracture><bone fracture healing><bone fracture repair><bone tissue formation><breast tumor cell><cancer cell><cell type><clinical care><compact bone><cortical bone><cytokine><diffused><diffuses><diffusing><diffusions><diphosphonate><drug treatment><experiment><experimental research><experimental study><experiments><femur fracture><fracture repair><global gene expression><global transcription profile><improved><in vivo><inflammatory breast cancer><inhibitor><kinase inhibitor><malignancy><malignant breast tumor><micro CT><micro computed tomography><microCT><microtomography><model of animal><neoplasm/cancer><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><normal ossification><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><ontogeny><ossification><osteochondral><osteochondral tissue><osteogenic><osteoprogenitor><osteoprogenitor cell><p44 MAPK><pre-clinical><preclinical><progenitor><protein blotting><radiation treatment><three dimensional><transcriptome><transcriptomics><translational therapeutics><translational therapy><treatment group><treatment with radiation>