Angela Vasaturo

Ultivue

Because of the poor prognosis of metastatic cancer, it is critical to determine exactly how different factors contribute to cancer spread. Mlecnik et al. examined the impact of tumor-intrinsic, microenvironmental, and immunological factors on tumor metastasis in colorectal cancer patients. They found that decreased presence of lymphatic vessels and...

Tumor infiltrating lymphocytes appear to be a predictor of survival in many cancers, including cutaneous melanoma. We applied automated multispectral imaging to determine whether density and distribution of T cells within primary cutaneous melanoma tissue correlate with survival of metastatic melanoma patients after dendritic cell (DC) vaccination....

Aims: The quality and quantity of the infiltration of immune cells into tumor tissues has substantial impact on patients' clinical outcome and is associated with response to immunotherapy. Therefore, the precise analysis of tumor-infiltrating lymphocytes (TIL) is becoming an important additional pathological biomarker. Analysis of TILs is usually...

Background: Despite the rapidly increasing number of new targeted and immunotherapeutic options over the past two decades, the prognosis of patients with NSCLC, even with early-stage tumors, is still poor and novel biomarkers are needed to better stratify patients in terms of survival and treatment response. A novel approach is to gain a holistic u...

Background: The advancement of AI in digital pathology is essential for analyzing image data containing billions of cells. This is true for digital pathology workflows using bright-field images, but this is even more critical for multiplex immunofluorescence (mIF) whole-slides due to the increased number of fluorescent channels and bit-depth per pi...

Background Characterization of the tumor microenvironment is becoming a clinical reality for discovering evidence of novel treatments’ efficacy. Multiplex immunofluorescence (mIF) assays have emerged as potent tools for immune-profiling analysis to examine spatial biology of tissues and determine tumor and immune cell interactions. Though, the succ...

Background Ultivue InSituPlex technology (ISP) enables whole-slide multiplex immune-fluorescence (mIF) staining of multiple protein targets on an autostainer and for conducting multiple rounds of ISP imaging and a terminal or initial H&E stain of the same slide. But to accurately phenotype each cell, one must co-register or ‘stack’ the images of ea...

Background Non-small-cell lung cancer (NSCLC) is the leading cause for cancer death. Current targeted- and immuno-therapies are effective in a patient subset, but causes for resistance and the complexity of the immune reaction are difficult to be identified on a single patient level. It is hypothesized that the tumor microenvironment (TME) and its...

Background: The Immunoscore (IS) is a quantitative digital pathology assay that evaluates the immune response in cancer patients. This study reports on the reproducibility of pathologists' visual assessment of CD3+- and CD8+-stained colon tumors, compared to IS quantification. Methods: An international group of expert pathologists evaluated 540...

Background Intratumoral (IT) myeloid dendritic cells (myDCs) play a pivotal role in initiating antitumor immune responses and relicensing of anti-tumor cytotoxic T lymphocytes within the tumor microenvironment. Talimogene laherparepvec (T-VEC) induces immunogenic cell death, thereby providing maturation signals and enhancing the release of tumor an...

Immunotherapy has transformed the treatment of metastatic and recurrent solid tumors. Advances in technology in the past few years have created unprecedented opportunities to identify biomarkers of disease processes, especially by using multi-omics technologies and datasets to derive valid and useful signatures of disease. Advances in multiplexed i...

Immunotherapy has transformed the treatment of metastatic and recurrent solid tumors. Advances in technology in the past few years have created unprecedented opportunities to identify biomarkers of disease processes, especially by using multi-omics technologies and datasets to derive valid and useful signatures of disease. Mouse tumor models are wi...

Conclusions In this study, we highlighted the benefits of using a combination of well-characterized TMAs, a fast, optimized 8-plex mIHC protocol, and a detailed analysis pipeline to characterize the immune-response in a broad range of cancer types and samples, leading to a better understanding of the TME as well as a streamlined workflow for furthe...

Progression of epithelial cancers predominantly proceeds by collective invasion of cell groups with coordinated cell–cell junctions and multicellular cytoskeletal activity. Collectively invading breast cancer cells express the gap junction protein connexin-43 (Cx43), yet whether Cx43 regulates collective invasion remains unclear. We here show that...

The tumour microenvironment (TME) forms a major obstacle in effective cancer treatment and for clinical success of immunotherapy. Conventional co-cultures have shed light onto multiple aspects of cancer immunobiology, but they are limited by the lack of physiological complexity. We develop a human organotypic skin melanoma culture (OMC) that allows...

Progression of epithelial cancers predominantly proceeds by collective invasion of cell groups with coordinated cell-cell junctions and multicellular cytoskeletal activity. Collectively invading breast cancer cells co-express adherens junctions and connexin-43 (Cx43) gap junctions in vitro and in patient samples, yet whether gap junctions contribut...

It is increasingly recognized that a deep characterization of the immune microenvironment is required for the identification of prognostic and predictive immune biomarkers. Recent advances in the field of tissue imaging resulted in the development of fluorescence multiplex IHC technologies enabling quantitative assessment of immune phenotypes and f...

Early detection and treatment are critical for improving the outcome of patients with cancer¹. Understanding the largely uncharted biology of carcinogenesis requires deciphering molecular processes in premalignant lesions, and revealing the determinants of the intralesional immune reaction during cancer development. The adaptive immune response wit...

Our recent understanding of the immune contribution to fight cancer has deeply modified the standard of care of cancer patients. As an example, immunotherapies by immune checkpoint inhibitors (ICI) anti-PD1/PDL1 are now approved in several cancer indications, such as Non-Small Cell Lung Cancers or melanoma. However, ICI are less effective for other...

Our recent understanding of the immune contribution to fight cancer has deeply modified the standard of care of cancer patients. As an example, immunotherapies by immune checkpoint inhibitors (ICI) anti-PD1/PDL1 are now approved in several cancer indications, such as Non-Small Cell Lung Cancers or melanoma. However, ICI are less effective for other...

The tumour microenvironment (TME) forms a major obstacle in effective cancer treatment and for clinical success of immunotherapy. Conventional co-cultures have shed light into multiple aspects of cancer immunobiology, but they are limited by the lack of physiological complexity. We developed a novel human, organotypic skin melanoma culture (OMC) th...

Treatment of metastatic colorectal cancer is based upon the assumption that metastases are homogeneous within a patient. We quantified immune cell types of 603 whole-slide metastases and primary colorectal tumors from 222 patients. Primary lesions, and synchronous and metachronous metastases, had a heterogeneous immune infiltrate and mutational div...

We examined how the immune microenvironment molds tumor evolution at different metastatic organs in a longitudinal dataset of colorectal cancer. Through multiplexed analyses, we showed that clonal evolution patterns during metastatic progression depend on the immune contexture at the metastatic site. Genetic evidence of neoantigen depletion was obs...

Figure S1. Flow cytometric analysis of intra‐tumoral lymphocytes from patients with colorectal cancerliver metastasis. A, Viable (aqua dye‐negative) leukocytes were gated based on CD45 expression. B, T cells were defined based on the expression of CD3. C, CD3+ T cells were further analyzed for CD4 and CD8. D, Percentages of Treg cells were then def...

Background and Objectives Patients with isolated colorectal‐cancer‐liver‐metastases (CRCLM) frequently undergo metastatectomy. Tumor‐infiltrating‐lymphocytes (TILs) have prognostic potential in the setting of primary colorectal cancer, however, their role in CRCLM is less studied. We aimed to study the spatial distribution and prognostic role of tu...

Background: The estimation of risk of recurrence for patients with colon carcinoma must be improved. A robust immune score quantification is needed to introduce immune parameters into cancer classification. The aim of the study was to assess the prognostic value of total tumour-infiltrating T-cell counts and cytotoxic tumour-infiltrating T-cells c...

Background: This study assesses how the metastatic immune landscape is impacting the response to treatment and the outcome of colorectal cancer (CRC) patients. Methods: Complete curative resection of metastases (n = 441) was performed for two patient cohorts (n = 153). Immune densities were quantified in the center and invasive margin of all met...

Platinum-based chemotherapeutics are amongst the most powerful anti-cancer drugs. Although their exact mechanism of action is not well understood, it is thought to be mediated through covalent DNA binding. We investigated the effect of platinum-based chemotherapeutics on signaling through signal transducer and activator of transcription (STAT) prot...

P15 Novel IL-2/mAb complexes mediate potent anti-tumor immunity which is augmented with anti-PD-1 mAb therapy Mark Rubinstein1, Kristina Andrijauskaite1, Marzena Swiderska-syn1, Kristin Lind2, Agnes Choppin2, Marina K Roell2 1Medical University of South Carolina, Charleston, SC, USA; 2XOMA Corporation, Berkeley, CA, USA Correspondence: Mark Rubin...

Melanoma is a highly malignant melanocyte-derived tumor and its incidence is increasing at outstanding rate. Despite different types of immunotherapy became available for the treatment of melanoma, including adoptive T cell transfer, immune checkpoint blockade, and vaccines such as dendritic cell vaccines, when applied to treat metastatic melanoma,...

Background Melanoma is a highly malignant melanocyte-derived tumor and its incidence is increasing at outstanding rate. Despite specific therapies have been explored for many years, no effective therapeutic options have been developed. Vaccination strategies, including Dendritic Cells (DC) based immunotherapy, are consistently increasing the propor...

Dendritic cell (DC) migration is essential for efficient host defense against pathogens and cancer, as well as for the efficacy of DC-based immunotherapies. However, the molecules that induce the migratory phenotype of DCs are poorly defined. Based on a large-scale proteome analysis of maturing DCs, we identified the GPI-anchored protein semaphorin...

Melanoma is a highly malignant melanocyte-derived tumor and its incidence is increasing at outstanding rate. Despite different types of immunotherapy became available for the treatment of melanoma, including adoptive T cell transfer, immune checkpoint blockade, and vaccines such as dendritic cell vaccines, when applied to treat metastatic melanoma,...

Antitumor immunity can be enhanced by the coordinated release and delivery of antigens and immunestimulating agents to antigen-presenting cells via biodegradable vaccine carriers. So far, encapsulation of TLR ligands and tumor-associated antigens augmented cytotoxic T cell (CTLs) responses. Here, we compared the efficacy of the invariant NKT (iNKT)...

Multispectral imaging is a novel microscopy technique that combines imaging with spectroscopy to obtain both quantitative expression data and tissue distribution of different cellular markers. Tetraspanins CD37 and CD53 are four-transmembrane proteins involved in cellular and humoral immune responses. However, comprehensive immunohistochemical anal...

Cancer cells evolve from normal cells throughout life and are usually recognized by our immune system and destroyed, a process called immunosurveillance. Unfortunately, in some instances cancer cells paralyze our immune system, resulting in outgrowth and spreading of the tumor. Understanding the complexity of immunomodulation by tumors is important...

The aim of therapeutic dendritic cell (DC) vaccines in cancer immunotherapy is to activate cytotoxic T cells to recognize and attack the tumor. T cell activation requires the interaction of the T cell receptor with a cognate major-histocompatibility complex-peptide complex. Although initiated by antigen engagement, it is the complex balance between...

The directional cell response to chemical gradients, referred to as chemotaxis, plays an important role in physiological and pathological processes including development, immune response and tumor cell invasion. Despite such implications, chemotaxis remains a challenging process to study under physiologically-relevant conditions in-vitro, mainly du...

Directional migration of freshly isolated human neutrophils in 3D collagen matrix towards Il-8. Images were taken at 10× magnification, and correspond to the Z projection of 5 layers reported at a reduced time and spatial resolution with about one frame every two minutes, in order to minimize the file dimension. The first 30 minutes, where each fra...

T-test on velocity module and components. Percentage of cells that rejected the null hypothesis at the 5% significance level (p<0.05), calculated for the velocity modulus and components, is reported. Y is the direction of the chemoattractant gradient. A and B panels are relative to data from donors A and B. (TIF)

Cell migration is dependent on the control of signaling events that play significant roles in creating contractile force and in contributing to wound closure. We evaluated wound closure in fibroblasts from mice overexpressing (TgPED) or lacking ped/pea-15 (KO), a gene overexpressed in patients with type 2 diabetes. Cultured skin fibroblasts isolate...

Cancer invasion into an extracellular matrix (ECM) results from a biophysical reciprocal interplay between the expanding cancer lesion and tissue barriers imposed by the adjacent microenvironment. In vivo, connective tissue provides both densely packed ECM barriers adjacent to channel/track-like spaces and loosely organized zones, both of which may...

An unresolved question in coeliac disease is to understand how some toxic gliadin peptides, in particular p31-43, can initiate an innate response and lead to tissue transglutaminase (TG2) upregulation in coeliac intestine and gliadin sensitive epithelial cell lines. Aim We addressed whether the epithelial uptake of p31-43 induces an intracellular p...

Cystic fibrosis (CF) is a monogenic disease caused by mutations in the CF transmembrane conductance regulator (CFTR) gene. CF is characterized by chronic bacterial lung infections and inflammation, and we have previously reported that tissue transglutaminase (TG2), a multifunctional enzyme critical to several diseases, is constitutively up-regulate...