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Mechanistic Insights and Diagnostic Applications for Hypoxia-Induced Vasorin in Pancreatic Cancer

缺氧诱导的 Vasorin 在胰腺癌中的机制见解和诊断应用

基本信息

批准号：
10088332
负责人：
Lisa Kirkemo
金额：
$ 4.11万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-01-01 至 2022-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10088332
关键词：
Antibodies Antigens Apoptotic Bacteriophages Binding Binding Proteins Biological Assay Biological Markers Biotinylation Cancer Cell Growth Cancer Diagnostics Cell Line Cell Proliferation Cell Survival Cell surface Cells Classification Cleaved cell Data Data Set Detection Development Diagnosis Diagnostic Disease Disease model Early Diagnosis Early Intervention Environmental Risk Factor Enzyme-Linked Immunosorbent Assay Epitopes Event Exhibits Fab Immunoglobulins Fibroblasts Future Genes Glioblastoma Goals Growth Hematologic Neoplasms Human Hypoxia Image Immune ImmunoPET Immunoprecipitation In Vitro Label Link Malignant Neoplasms Malignant neoplasm of pancreas Mass Spectrum Analysis Membrane Metabolic Methods Modality Modeling Molecular Target N-terminal NOTCH1 gene NU/NU Mouse Neoplasm Metastasis Non-Invasive Cancer Detection Oxidative Stress Oxygen Pancreas Pathway interactions Patients Phage Display Phenotype Play Positron-Emission Tomography Primary Neoplasm Proteins Proteolysis Proteome Proteomics Publishing Recombinant Antibody Recombinant Proteins Recombinants Role Serum Signal Transduction Site Solid Solid Neoplasm Stem Cell Development Streptavidin Stress Surface Technology Testing Therapeutic Therapeutic Intervention Tissues Toxic effect Western Blotting Zirconium antibody diagnostic antibody engineering base cancer biomarkers cancer cell cancer immunotherapy cancer stem cell cancer type cell growth clinically relevant deprivation desensitization design experience experimental study glycoproteomics in vivo in vivo Model insight knock-down migration mortality mouse model novel novel strategies pancreatic cancer cells pancreatic cancer model pancreatic cancer patients pancreatic neoplasm programs protective effect protein expression protein protein interaction radiotracer response tool transcriptomics tumor tumor hypoxia tumor microenvironment virtual

项目摘要

Project Summary With the advent of cancer immunotherapies, patients have seen dramatic increases in the availability of treatment options for various malignancies. However, many of these modalities lack efficacy in treating solid tumors, namely because of the unique set of environmental factors that exist within these tumors. One of the most ubiquitous features within the tumor microenvironment (TME) is oxygen deprivation, also known as hypoxia. Hypoxia has long been linked to increased metastasis and poorer prognoses for patients. In response to hypoxic stress, cancer cells activate highly regulated cellular pathways and gene programs that promote survival, migration, immune privilege, and increased mortality for patients. Across all solid tumors, pancreatic cancer exhibits the most severe hypoxic phenotype, which may contribute to the high mortality rate associated with this cancer type. These hypoxia-specific downstream effects suggest that tumor hypoxia can be used to elucidate selective biomarkers of solid pancreatic cancer for diagnostic and therapeutic purposes. Here, I utilize cell surface proteomics to identify a novel hypoxia-regulated target in pancreatic cancer called vasorin (VASN), a target previously implicated in the progression of glioblastoma. Furthermore, I have shown that VASN plays an important role in the growth and survival of pancreatic cancer under hypoxic stress. Finally, I have isolated and expressed ten unique antibody clones against the ectodomain of VASN. Using these antibodies, I found that VASN undergoes numerous cleavage events under hypoxia in vitro, which are unique from previously published results. We hypothesize that VASN cleavage is necessary for the survival and proliferation of pancreatic cancer under hypoxia, and that classification of expression levels for cleavage-specific forms of VASN in relevant tumor models will aid the downstream development of antibody-based diagnostics for hypoxic pancreatic cancer. To test this hypothesis, I will employ proteomic characterization and recombinant protein expression to identify the in vitro cellular consequences of VASN proteolysis. In order to isolate clones towards the membrane-retained form of cleaved VASN, I will use an established phage display-based recombinant antibody strategy. A selective antibody-based biotinylation strategy will be implemented to identify novel interacting partners of the membrane- retained form of cleaved VASN that may play an important role in downstream signaling. Finally, I will functionalize my existing and future anti-VASN antibody clones for serum-based ELISA detection and radiotracer-based immunoPET imaging of VASN, respectively, which will validate the presence of proteolyzed VASN in human serum and tumors, as well as in an in vivo nu/nu mouse model of pancreatic cancer. The proposed studies will provide mechanistic insights into the functional consequences of VASN proteolysis in pancreatic cancer, as well as providing evidence for the development of non-invasive, early-detection diagnostic alternatives for a highly metastatic and deadly disease.

项目总结