A5033741912- Peptidase Inhibition and Analysis
- Cancer, Hypoxia, and Metabolism
- Monoclonal and Polyclonal Antibodies Research
- Protease and Inhibitor Mechanisms
- Radiopharmaceutical Chemistry and Applications
- Metabolism, Diabetes, and Cancer
- Ubiquitin and proteasome pathways
- Medical Imaging Techniques and Applications
- Fibroblast Growth Factor Research
- Cell Adhesion Molecules Research
- Glycosylation and Glycoproteins Research
- Nanoparticle-Based Drug Delivery
- Cancer Research and Treatments
- Immune cells in cancer
- Chemical Synthesis and Analysis
- Protein Hydrolysis and Bioactive Peptides
- Prostate Cancer Treatment and Research
- CAR-T cell therapy research
- Nanoplatforms for cancer theranostics
- SARS-CoV-2 and COVID-19 Research
- Virus-based gene therapy research
- Immunotherapy and Immune Responses
- Computational Drug Discovery Methods
- Advanced Biosensing Techniques and Applications
- Click Chemistry and Applications
University of Wisconsin–Madison
2021-2025
University of Wisconsin Carbone Cancer Center
2024
Wisconsin Division of Public Health
2023
University of Minnesota Medical Center
2014-2021
University of Minnesota
2019-2021
University of California, San Francisco
2010-2015
Lawrence Berkeley National Laboratory
2013
Laboratoire d’Imagerie Biomédicale
2013
Johns Hopkins Medicine
2008-2009
Johns Hopkins University
2005-2009
Abstract Fibroblast-Activation Protein-α (FAP) is a membrane-bound serine protease that expressed on the surface of reactive stromal fibroblasts present within majority human epithelial tumors but not by normal tissues. FAP postprolyl peptidase differs from other dipeptidyl prolyl peptidases such as diprolylpeptidase 4 in it also has gelatinase and collagenase endopeptidase activity. Therefore, represents potential pan-tumor target whose enzymatic activity can be exploited for intratumoral...
Abstract Components of the plasminogen activation system, which are overexpressed in aggressive breast cancer subtypes, offer appealing targets for development new diagnostics and therapeutics. By comparing gene expression data patient populations cultured cell lines, we identified elevated levels urokinase receptor (uPAR, PLAUR) highly subtypes lines. Recombinant human anti-uPAR antagonistic antibodies exhibited potent binding vitro to surface cells expressing uPAR. In vivo these detected...
Metastatic castration-resistant prostate cancer (mCRPC) is a lethal, heterogeneous disease with few therapeutic strategies that significantly prolong survival. Innovative therapies for mCRPC are needed; however, the development of new relies on accurate imaging to assess metastasis and monitor response. Standard modalities require improvement there remains need selective sensitive probes can be widely used in patients mCRPC.We evaluated transmembrane protease fibroblast activation protein...
Combating the COVID-19 pandemic requires potent and low-cost therapeutics. We identified a series of single-domain antibodies (i.e., nanobody),
Abstract Adeno-associated virus (AAV) has been remarkably successful in the clinic, but its broad tropism is a practical limitation of precision gene therapy. A promising path to engineer AAV addition binding domains capsid that recognize cell surface markers present on targeted type. We have recently identified two previously unexplored regions near 2/5-fold wall and 5-fold pore are amenable insertion larger protein domains, including nanobodies. Here, we demonstrate these hotspots...
Interactions between urokinase plasminogen activator receptor (uPAR) and its various ligands regulate tumor growth, invasion, metastasis. Antibodies that bind specific uPAR epitopes may disrupt these interactions, thereby inhibiting processes. Using a highly diverse naïve human fragment of the antigen binding (Fab) phage display library, we identified 12 unique Fabs uPAR. Two antibodies compete against (uPA) for binding, whereas third competes with beta1 integrins binding. These competitive...
Single-domain Variable New Antigen Receptors (VNARs) from the immune system of sharks are smallest naturally occurring binding domains found in nature. Possessing flexible paratopes that can recognize protein motifs inaccessible to classical antibodies, VNARs have yet be exploited for development SARS-CoV-2 therapeutics. Here, we detail identification a series VNAR phage display library screened against receptor domain (RBD). The ability neutralize pseudotype and authentic live virus...
Cancer-associated fibroblasts (CAFs) in the stroma of solid tumors promote an immunosuppressive tumor microenvironment (TME) that drives resistance to therapies. The expression protease fibroblast activation protein (FAP) on surface CAFs has made FAP a target for development therapies dampen immunosuppression. Relatively few biologics have been developed and none exploit unique engagement properties Variable New Antigen Receptors (VNARs) from shark antibodies. As smallest binding domain...
Abstract Background: The mesenchymal transcript factors is a receptor tyrosine kinase that when dysregulated or overexpressed can stimulate oncogenesis. Alterations in MET are found 5% of non-small cell lung cancer but also head and neck cancers resistant to inhibitors targeting other oncogenic drivers. Our objective evaluate novel, camelid-derived, binding antibody as potential theranostic agent used expressing cancers. Methods: anti-MET VHH camelid antibody, 1E7-Fc, was identified from...
The M1 family of metalloproteases represents a large number exopeptidases that cleave single amino acid residues from the N-terminus peptide substrates. One member this has been well studied is aminopeptidase N (APN), multifunctional protease known to biologically active peptides and aide in coronavirus entry. proteolytic activity APN promotes cancer angiogenesis metastasis making it an important target for therapy. To understand substrate specificity development targeted inhibitors, we used...
Subtype-targeted therapies can have a dramatic impact on improving the quality and quantity of life for women suffering from breast cancer. Despite an initial therapeutic response, cancer recurrence acquired drug-resistance are commonplace. Non-invasive imaging probes that identify drug-resistant lesions urgently needed to aid in development novel drugs effective utilization established The protease receptor urokinase plasminogen activator (uPAR) is target be exploited non-invasive imaging....
Abstract Functional imaging of proteolytic activity is an emerging strategy to quantify disease and response therapy at the molecular level. We present a new peptide-based probe technology that advances these goals by exploiting enzymatic deposit probes labelled with near-infrared (NIR) fluorophores or radioisotopes in cell membranes disease-associated proteolysis. This allows for non-invasive detection protease vivo ex tracking deposited tissues. demonstrate thrombin generation murine model...
The Mesenchymal Epithelial Transition (MET) receptor tyrosine kinase is upregulated or mutated in 5% of non-small-cell lung cancer (NSCLC) patients and overexpressed multiple other cancers. We sought to develop a novel single-domain camelid antibody with high affinity for MET that could be used deliver conjugated payloads expressing From naïve variable-heavy-heavy (VHH) domain phage display library, we identified VHH clone termed 1E7 displayed human was cross-reactive across species. When...
Cancer-associated fibroblasts (CAF) are a prominent cell type within the tumor microenvironment (TME) where they known to promote cancer growth and survival, angiogenesis, drug resistance, immunosuppression. The transmembrane prolyl protease fibroblast activation protein (FAP) is expressed on surface of highly protumorigenic CAFs found in stroma nearly every epithelial origin. widespread expression FAP has made it an attractive therapeutic target based underlying hypothesis that eliminating...
In colorectal cancer (CRC), attempts to identify cell-specific markers guide antibody-mediated therapeutics have failed uncover that are both exclusive tissues and abundant across CRCs. Alternatively, cancer-associated fibroblasts (CAFs), which in the tumor microenvironment upregulate unique surface markers, not found healthy tissues. Here, we evaluated expression patterns of CAF-associated proteins α-smooth muscle actin (αSMA), fibroblast activation protein (FAP), podoplanin (PDPN), matrix...
Prostate-specific antigen (PSA), a serine protease belonging to the human kallikrein family, is best known as prostate cancer biomarker. Emerging evidence suggests that PSA may also play salient role in development and progression. With large amounts of enzymatically active continuously selectively produced by all stages cancer, an attractive target. inhibitors, therefore, represent promising class therapeutics and/or imaging agents. displays chymotrypsin-like specificity, cleaving after...
Large changes in the activity of serotonin N-acetyltransferase (arylalkylamine N-acetyltransferase, AANAT) pineal gland control rhythmic production time-keeping hormone melatonin. The AANAT reflects amount and activation state protein, both which increase at night. molecular basis this regulation is now becoming known, recent data indicate that involves phosphorylation-dependent binding to 14-3-3 protein two sites, one which, Ser-205, located several residues from C terminus. In study, we...
Abstract The increased proteolytic activity of membrane-bound and secreted proteases on the surface cancer cells in transformed stroma is a common characteristic aggressive metastatic prostate cancer. We describe here development an active site-specific probe for detecting peritumoral protease expressed by surrounding tumor microenvironment. Using human fragment antigen-binding phage display library, we identified antibody termed U33 that selectively inhibited form urokinase plasminogen...
Abstract Combating the COVID-19 pandemic requires potent and low-cost therapeutics. We identified a novel series of single-domain antibodies (i.e., nanobody), Nanosota-1, from camelid nanobody phage display library. Structural data showed that Nanosota-1 bound to oft-hidden receptor-binding domain (RBD) SARS-CoV-2 spike protein, blocking out viral receptor ACE2. The lead drug possessing an Fc tag ( Nanosota-1C-Fc ) RBD with K d 15.7picomolar (∼3000 times more tightly than ACE2 did) inhibited...
Abstract Metastatic castration-resistant prostate cancer (mCRPC) has been largely resistant to immunotherapy. Natural killer (NK) cells are cytotoxic lymphocytes that detect and kill transformed without prior sensitization, their infiltration into tumors corresponds with an increased overall survival among patients mCRPC. We sought harness this knowledge develop approach NK-cell based immunotherapy for engineered NK cell line (NK-92MI) express CD64, the sole human high-affinity IgG Fcγ...
Here, we document the discovery of a monoclonal antibody that selectively binds to both human and murine fibroblast activation protein alpha (FAP), serine protease is overexpressed on cancer-associated fibroblasts (CAFs), making it an attractive therapeutic target for aiding abetting tumor microenvironment. The lead antibody, B12, was identified from naïve single-chain variable fragment phage display library screened against recombinant FAP magnetic beads. heavy light chains B12 were cloned...
Prostate Specific Antigen's (PSA) role as a biomarker for prostate cancer is well established but the physiological of its serine protease activity in pathobiology normal and carcinogenesis remains largely unknown. In light recent studies that implicate PSA's enzymatic initiation and/or progression cancer, we performed molecular modeling study substrate binding at catalytic site PSA wherein PSA-selective (HSSKLQ) was docked an acyl-enzyme conformation to three-dimensional homology model PSA....
A common hallmark of cancers with highly aggressive phenotypes is increased proteolysis in the tumor and surrounding microenvironment. Prostate cancer has a number proteases uniquely associated it that may play various important roles disease progression. In this report, we utilize peritumoral proteolytic activity prostate to activate engineered peptide constructs for treatment noninvasive imaging cancer. Using modular "propeptide" approach, cationic diastereomeric pore-forming domain was...