Publications

CRISPR-Cas9 gene editing of hepatitis B virus in chronically infected humanized mice

Daniel Stone,et al, Molecular Therapy Methods & Clinical Development, 2021
Hepatitis B leads to over 850,000 deaths annually due to complications from chronic infection including cirrhosis and hepatocellular carcinoma. While antiviral drugs successfully reduce viral loads in patients, a cure does not yet exist. This study out of the Jerome laboratory at the Fred Hutchinson Cancer Research Center performed in vivo gene editing utilizing a mouse model for chronic HBV infections and demonstrated a proof-of-concept of safe and effective gene editing of the genomic form responsible for replication and persistence. This study utilized the HALO Link image management platform in conjunction with image acquisition on a Leica Biosystems Aperio VERSA 200 slide scanner for their RNAscope studies of Cas9 and HBV.

CRISPR-Cas9 gene editing of hepatitis B virus in chronically infected humanized mice Read More »

Monomeric/dimeric forms of Fgf15/FGF19 show differential activity in hepatocyte proliferation and metabolic function

Courtney M. Williams et al, The FASEB Journal, 2021
Williams and colleagues at Regeneron Pharmaceuticals perform structure/function relationship studies on Fgf15 and FGF19 using site-directed mutagenesis and downstream functional assays in order to understand their distinct functions in a common pathway. Both molecules are therapeutic targets due to involvement in hepatocellular carcinoma and bile acid production. This publication identifies a single cysteine residue is identified that controls dimerization and hepatocyte proliferation. Understanding these molecular pathways may inform future studies on hepatocellular carcinoma while limiting toxicity and induction of hepatocyte proliferation. Immunohistochemistry images were quantified in HALO Link using the Cytonuclear IHC module.

Monomeric/dimeric forms of Fgf15/FGF19 show differential activity in hepatocyte proliferation and metabolic function Read More »

Multiomic analysis and immunoprofiling reveal distinct subtypes of human angiosarcoma

Jason Yongsheng Chan,et al, The Journal of Clinical Investigation, 2020.
To molecularly characterize the rare and aggressive tumors known as angiosarcomas, Chan et al utilized a combination of transcriptomic immuno-oncology profiling with multiplex immunohistochemistry and immunofluorescence. HALO image analysis software was used for the quantification of PD-L1, CD68, CD8, FOXP3, CD15, and ERG. From these studies, three phenotypes of patients were identified according to etiology, anatomical origin, signaling pathways, and the tumor inflammation signature. These distinct biological phenotypes provide opportunity for future studies to improve prognosis and treatment of a rare and deadly disease.

Multiomic analysis and immunoprofiling reveal distinct subtypes of human angiosarcoma Read More »

PNOCARC Neurons Promote Hyperphagia and Obesity upon High-Fat-Diet Feeding

Alexander Jaisl, et al, Neuron, 2020
To advance the molecular understanding of obesity, Jais and colleagues set out to identify neuronal populations in mice that respond to being fed a calorie dense, highly palatable food. They identified prepronociceptin (PNOC)-expressing neurons in the hypothalamus that are activated upon acute high fat diet feeding and promote overconsumption. In this publication, HALO image analysis software was used for quantification of transgene expression in mouse neuronal tissue. Future studies on PNOC-expressing neurons may advance prevention and treatment of obesity and associated metabolic diseases.

PNOCARC Neurons Promote Hyperphagia and Obesity upon High-Fat-Diet Feeding Read More »

An ex vivo tumor fragment platform to dissect response to PD-1 blockade in cancer

Paula Voabil,et al, Nature Medicine, 2021.
Dr. Paula Voabil and colleagues used a patient derived tumor fragment platform to investigate immune responses immediately following PD-1 blockade. The Tissue Classifier Add-on and the Multiplex IHC module of HALO® were used in this publication to quantify immune biomarkers in the tumor and stroma, contributing to their conclusion that the ability of tumors to respond to PD-1 blockage correlates with the ability of intratumoral immune cells to be reactivated by the blockade.

An ex vivo tumor fragment platform to dissect response to PD-1 blockade in cancer Read More »

Neoadjuvant immunotherapy leads to pathological responses in MMR-proficient and MMR-deficient early-stage colon cancers

Myriam Chalabi, et al, Nature Medicine, 2020.
Immunotherapy is known to be effective in late-stage, mismatch repair (MMR) deficient colorectal cancers but not in MMR proficient cancer. This study reports the early results from the NICHE clinical trial that is evaluating treatment of early stage, nonmetastatic preoperative colon cancer with a CTLA-4 inhibitor and a PD-1 inhibitor. Chalabi and colleagues report that the treatment was well tolerated and that a major pathological response was seen in 19/20 patients with MMR deficient tumors and 3/15 MMR proficient tumors. HALO image analysis software was used in this publication for analysis of T-cell biomarkers CD3, CD8, and FOXP3 on chromogenically stained tumor biopsies. First, image registration with HALO was performed of three serial tissue sections. Tissue sections were annotated manually, and quantification of DAB positivity was performed with the Multiplex IHC module and expressed as a function of tumor area. The authors conclude that immunotherapy shows promise to become the standard of care in a defined group of early-stage colon cancers.

Neoadjuvant immunotherapy leads to pathological responses in MMR-proficient and MMR-deficient early-stage colon cancers Read More »

Transmission of tauopathy strains is independent of their isoform composition

Alexander Jaisl, et al, Neuron, 2020.
To advance the molecular understanding of obesity, Jais and colleagues set out to identify neuronal populations in mice that respond to being fed a calorie dense, highly palatable food. They identified prepronociceptin (PNOC)-expressing neurons in the hypothalamus that are activated upon acute high fat diet feeding and promote overconsumption. In this publication, HALO image analysis software was used for quantification of transgene expression in mouse neuronal tissue. Future studies on PNOC-expressing neurons may advance prevention and treatment of obesity and associated metabolic diseases.

Transmission of tauopathy strains is independent of their isoform composition Read More »

Mechanical regulation of glycolysis via cytoskeleton architecture

Jin Suk Park,et al, Nature, 2020.
Park and colleagues explored the question of how regulation of glycolysis responds to structural changes in tissue architecture and chose lung cells and tissue for their studies due to the regular mechanical stress experienced during respiration. In vitro studies demonstrated downregulation of glycolysis following a change in substrate via the degradation of the platelet isoform of the phosphofructokinase (PFKP) enzyme. In vitro studies also showed that oncogenic transformation changed the ability of PFKP expression to change in response to mechanical stress. The researchers corroborated this result with in vivo work by performing immunohistochemical assays on control and malignant tissue cores from bronchi of lung cancer patients. The HALO Tissue Classifier Add-on was used to classify epithelium, stroma, and tumor within the tissue cores and the Cytonuclear IHC module was used to quantify cytoplasmic PFKP staining in bronchial epithelium.

Mechanical regulation of glycolysis via cytoskeleton architecture Read More »

Association of COVID-19 inflammation with activation of the C5a–C5aR1 axis

Julien Carvelli, et al, Nature, 2020
Researchers aiming to block excessive lung inflammation in COVID-19 patients found upregulated immune checkpoint biomarkers in patients with a range of COVID-19 symptoms (from paucisymptomatic to acute respiratory distress syndrome). In addition, Carvelli et al found increased expression of C5a, an inflammatory mediator, in serum and the C5aR1 receptor on myeloid cells in COVID-19 patients, which are known to initiate inflammatory responses by recruiting neutrophils and monocytes to lungs. An in vitro neutrophil migration assay quantified with the CytoNuclear FL Module of HALO demonstrated that the clinical stage therapeutic monoclonal antibody, avdoralimab, effectively inhibited C5a-induced neutrophil migration. The authors propose use of avdoralimab to limit excessive lung inflammation associated with acute respiratory distress in COVID-19 patients.

Association of COVID-19 inflammation with activation of the C5a–C5aR1 axis Read More »

Natural killer cells in the human lung tumor microenvironment display immune inhibitory functions

Jules Russick, et al, Journal for ImmunoTherapy of Cancer, 2020.
Natural killer (NK) cells are known to have cytotoxic effector functions in tumor immunosurveillance. More recently, evidence of a second potentially inhibitory or regulatory role have emerged. Here, Russick et al compared expression patterns of NK cells inside tumors to nontumoral NK cells to understand their inhibitory functions in the context of the non-small cell lung carcinoma (NSCLC) tumor microenvironment (TME). These studies identified a novel and specific gene signature of NK cells dysregulation in NSCLC that suggests that the TME may induce suppressive NK cells. HALO image analysis software was leveraged to quantify CD8 expression of formalin fixed paraffin embedded NSCLC sections with CD8 immunohistochemistry and a hematoxylin counterstain. While the presence of CD8 was previously known to be associated with good clinical outcomes, Russick and colleagues uncovered a more complex relationship where patients with low CD8+ T cells and high NK cell density had good outcomes, and those with high CD8+ T cells and high NK cell density had poor outcomes.

Natural killer cells in the human lung tumor microenvironment display immune inhibitory functions Read More »

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