A5030924932- Cancer, Hypoxia, and Metabolism
- Metabolism, Diabetes, and Cancer
- Peptidase Inhibition and Analysis
- Medical Imaging Techniques and Applications
- Fibroblast Growth Factor Research
- Ubiquitin and proteasome pathways
- Cardiomyopathy and Myosin Studies
- Cardiovascular Effects of Exercise
- Immune cells in cancer
- Medical Imaging and Pathology Studies
- Radiopharmaceutical Chemistry and Applications
- Lung Cancer Research Studies
- Radiomics and Machine Learning in Medical Imaging
- Microtubule and mitosis dynamics
- Genetic Neurodegenerative Diseases
- Glycosylation and Glycoproteins Research
- Cardiac electrophysiology and arrhythmias
- Protease and Inhibitor Mechanisms
University of Wisconsin–Madison
2024
James Madison University
2019-2021
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...
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...
Desmoplakin (DSP) is a large (~260 kDa) protein found in the desmosome, subcellular complex that links cytoskeleton of one cell to its neighbor. A mutation ‘hot-spot’ within NH2-terminal third DSP (specifically, residues 299–515) associated with both cardiomyopathies and skin defects. In select variants, disease linked specifically uncovering previously-occluded calpain target site (residues 447–451). Here, we partially stabilize these calpain-sensitive clinical variants through addition...
<div>Abstract<p>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...
<p>Downstream effects of huB12-MMAE treatment on gene expression in CAF-tumor models Stacks. <b>A,</b> Concentration selected factors supernatant monoculture and coculture conditions. <b>B,</b> Relative mRNA select genes hPrCSC-44 cells treated untreated <b>C,</b> 22Rv1 Data for B C expressed as normalized relative quantity (NRQ). *, <i>P</i> < 0.05.</p>
<p>Specificity of huB12-MMAE treatment in the Stacks. <b>A,</b> Workflow and analysis CAF-tumor models using huB12-MMAE. <b>B,</b> Viability CAFS (hPrCSC-44/top) tumor cells (22Rv1/bottom) monoculture (left) coculture (right) indicated concentrations for 72 hours.</p>
<p>mRNA expression of cytokines</p>
<p>Specificity of huB12-MMAE treatment in the Stacks. <b>A,</b> Workflow and analysis CAF-tumor models using huB12-MMAE. <b>B,</b> Viability CAFS (hPrCSC-44/top) tumor cells (22Rv1/bottom) monoculture (left) coculture (right) indicated concentrations for 72 hours.</p>
<p>ADC efficacy in multiple cell lines</p>
<p><i>In vitro</i> and <i>in vivo</i> therapeutic efficacy of huB12-MMAE. <b>A,</b> FAP-targeted ADCs show a potent FAP-selective cytotoxic activity. Cell viability hPrCSC-44, CWR-R1FAP, PC3, CWR-R1 cell lines. Cells were treated for 72 hours with serial dilution either huB12 IgG or an isotype control without MMAE warhead. Data are represented as mean ± SEM (<i>n</i> = 3 biological replicates). <b>B,</b> HuB12 ADC therapy...
<p>Downstream effects of huB12-MMAE treatment on gene expression in CAF-tumor models Stacks. <b>A,</b> Concentration selected factors supernatant monoculture and coculture conditions. <b>B,</b> Relative mRNA select genes hPrCSC-44 cells treated untreated <b>C,</b> 22Rv1 Data for B C expressed as normalized relative quantity (NRQ). *, <i>P</i> < 0.05.</p>
<p><i>In vitro</i> and <i>in vivo</i> therapeutic efficacy of huB12-MMAE. <b>A,</b> FAP-targeted ADCs show a potent FAP-selective cytotoxic activity. Cell viability hPrCSC-44, CWR-R1FAP, PC3, CWR-R1 cell lines. Cells were treated for 72 hours with serial dilution either huB12 IgG or an isotype control without MMAE warhead. Data are represented as mean ± SEM (<i>n</i> = 3 biological replicates). <b>B,</b> HuB12 ADC therapy...
<p>VEGFA expression</p>
<p>Proliferation and survival marker gene expression</p>
<div>Abstract<p>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...
<p>Determining the therapeutic window in Stacks</p>
<p>HuB12 internalizes into FAP-expressing cells. <b>A,</b> Confocal microscopy images of hPrCSC-44 cells (top) or PC3 (bottom) after incubation with Alexa Fluor-647 (AF647)-labeled huB12 (50 nmol/L) for 1 hour. Single-channel huB12-AF647 localization, fluorescein-dextran labeled endosomes, and CellBrite Orange-labeled plasma membrane are shown. Composite depicting whole-cell enlarged regions interest shown as colored fluorescence overlays. <b>B,</b>...
<p>VEGFA expression</p>
<p>PET/CT imaging of FAP <i>in vivo</i>. <b>A,</b> Representative PET/CT images mice bearing subcutaneous 22Rv1/hPrCSC-44 xenografts (white arrowhead). Mice (<i>n</i> = 3/group) received 3.5 MBq (25–50 µg, 7.7 µg/MBq) [<sup>89</sup>Zr]Zr-HuB12, 3.6 7.5 [<sup>89</sup>Zr]Zr-F19 or 3.4 7.9 [<sup>89</sup>Zr]Zr-J591 via tail vein and were imaged at the indicated timepoints. <b>B,</b> Quantitative analysis...
<p>ADC efficacy in multiple cell lines</p>
<p>PET/CT imaging of FAP <i>in vivo</i>. <b>A,</b> Representative PET/CT images mice bearing subcutaneous 22Rv1/hPrCSC-44 xenografts (white arrowhead). Mice (<i>n</i> = 3/group) received 3.5 MBq (25–50 µg, 7.7 µg/MBq) [<sup>89</sup>Zr]Zr-HuB12, 3.6 7.5 [<sup>89</sup>Zr]Zr-F19 or 3.4 7.9 [<sup>89</sup>Zr]Zr-J591 via tail vein and were imaged at the indicated timepoints. <b>B,</b> Quantitative analysis...
<p>Determining the therapeutic window in Stacks</p>
<p>Antibody Humanization</p>
<p>HuB12 internalizes into FAP-expressing cells. <b>A,</b> Confocal microscopy images of hPrCSC-44 cells (top) or PC3 (bottom) after incubation with Alexa Fluor-647 (AF647)-labeled huB12 (50 nmol/L) for 1 hour. Single-channel huB12-AF647 localization, fluorescein-dextran labeled endosomes, and CellBrite Orange-labeled plasma membrane are shown. Composite depicting whole-cell enlarged regions interest shown as colored fluorescence overlays. <b>B,</b>...
<p>Proliferation and survival marker gene expression</p>