88 results on '"Bzorek M"'
Search Results
2. 440 The Proteome of microdissected nevi- and melanoma cells reveal upregulation of Hippo-YAP and PI3K-mTOR pathway
- Author
-
Naimy, S., primary, Kuczek, D., additional, Bzorek, M., additional, Løvendorf, M.B., additional, Litman, T., additional, Mund, A., additional, Gjerdrum, L.R., additional, Mann, M., additional, and Dyring-Andersen, B., additional
- Published
- 2022
- Full Text
- View/download PDF
3. The Vitamin D Analogue Calcipotriol Reduces the Frequency of CD8+IL-17+ T Cells in Psoriasis Lesions
- Author
-
Dyring-Andersen, B., Bonefeld, C. M., Bzorek, M., Lvendorf, M. B., Lauritsen, J. P. H., Skov, L., and Geisler, C.
- Published
- 2015
- Full Text
- View/download PDF
4. Role of B-cells in Mycosis Fungoides
- Author
-
Nielsen, P, primary, Eriksen, J, additional, Sørensen, M, additional, Wehkamp, U, additional, Lindahl, L, additional, Bzorek, M, additional, Iversen, L, additional, Woetman, A, additional, Ødum, N, additional, Litman, T, additional, and Gjerdrum, L, additional
- Published
- 2021
- Full Text
- View/download PDF
5. Expression of anaplastic lymphoma kinase (ALK) in merkel cell carcinomas: O4-7
- Author
-
Filtenborg-Barnkob, B E and Bzorek, M
- Published
- 2013
- Full Text
- View/download PDF
6. Th1/Th17 are the main effector cells in human nickel allergy: FS12.05
- Author
-
Dyring-Andersen, B, Skov, L, Løvendorf, M, Bzorek, M, Søndergaard, K, Lauritsen, J P, Dabelsteen, S, Geisler, C, and Bonefeld, C M
- Published
- 2012
7. TWEAK receptor Fn14 in severe inflammatory liver disease - progenitor cell or remodelling marker?: O3-5
- Author
-
Olsen, TE, Bzorek, M, and Vainer, B
- Published
- 2012
8. The human placenta from heavy smokers: evaluation of vasoactive peptides by immunohistochemistry
- Author
-
CLAUSEN, H. V., LARSEN, L. GRUPE, JØRGENSEN, A., and BZOREK, M.
- Published
- 2007
9. Clinical and Histological Characteristics of Mycosis Fungoides and Sézary Syndrome: A Retrospective, Single-centre Study of 43 Patients from Eastern Denmark
- Author
-
Nielsen, P, primary, Eriksen, J, additional, Wehkamp, U, additional, Lindahl, L, additional, Gniadecki, R, additional, Fogh, H, additional, Fabricius, S, additional, Bzorek, M, additional, Ødum, N, additional, and Gjerdrum, L, additional
- Published
- 2019
- Full Text
- View/download PDF
10. A diagnostic two gene classifier in patients with mycosis fungoides: a retrospective multicenter study
- Author
-
Nielsen, P.R., primary, Eriksen, J.O., additional, Lindahl, L.M., additional, Wehkamp, U., additional, Andersen, G.R., additional, Bzorek, M., additional, Litman, T., additional, Ødum, N., additional, and Gjerdrum, L.M.R., additional
- Published
- 2019
- Full Text
- View/download PDF
11. Endometrial immune markers are potential predictors of normal fertility and pregnancy success
- Author
-
Kofod, L.M., primary, Lindhard, A., additional, Bzorek, M., additional, Eriksen, Jens Ole, additional, Larsen, L.G., additional, and Hviid, T.V.F., additional
- Published
- 2018
- Full Text
- View/download PDF
12. Interleukin (IL)-17A and IL-22-producing neutrophils in psoriatic skin
- Author
-
Dyring-Andersen, B., primary, Honoré, T. V., additional, Madelung, A., additional, Bzorek, M., additional, Simonsen, S., additional, Clemmensen, S. N., additional, Clark, R. A., additional, Borregaard, N., additional, and Skov, L., additional
- Published
- 2017
- Full Text
- View/download PDF
13. 630 Neutrophils are a major source of IL-17A in psoriatic skin and can produce IL-17F and IL-22
- Author
-
Dyring-Andersen, B., primary, Velte Honoré, T., additional, Madelung, A., additional, Bzorek, M., additional, Simonsen, S., additional, Novrup Clemmensen, S., additional, Clark, R.A., additional, Borregaard, N., additional, and Skov, L., additional
- Published
- 2017
- Full Text
- View/download PDF
14. 240 IL-17A- and IL-22-producing neutrophils in psoriatic skin
- Author
-
Dyring-Andersen, B., primary, Velte Honoré, T., additional, Madelung, A., additional, Bzorek, M., additional, Simonsen, S., additional, Novrup Clemmensen, S., additional, Borregaard, N., additional, and Skov, L., additional
- Published
- 2016
- Full Text
- View/download PDF
15. The Vitamin D Analogue Calcipotriol Reduces the Frequency of CD8+IL-17+T Cells in Psoriasis Lesions
- Author
-
Dyring-Andersen, B., primary, Bonefeld, C. M., additional, Bzorek, M., additional, Løvendorf, M. B., additional, Lauritsen, J. P. H., additional, Skov, L., additional, and Geisler, C., additional
- Published
- 2015
- Full Text
- View/download PDF
16. Human leukocyte antigen-G in the male reproductive system and in seminal plasma
- Author
-
Larsen, M. H., primary, Bzorek, M., additional, Pass, M. B., additional, Larsen, L. G., additional, Nielsen, M. W., additional, Svendsen, S. G., additional, Lindhard, A., additional, and Hviid, T. V. F., additional
- Published
- 2011
- Full Text
- View/download PDF
17. Correlation between Thymidylate Synthase Gene Variants, RNA and Protein Levels in Primary Colorectal Adenocarcinomas
- Author
-
Kristensen, MH, primary, Weidinger, M, additional, Bzorek, M, additional, Pedersen, PL, additional, and Mejer, J, additional
- Published
- 2010
- Full Text
- View/download PDF
18. The Vitamin D Analogue Calcipotriol Reduces the Frequency of CD8+ IL-17+ T Cells in Psoriasis Lesions.
- Author
-
Dyring‐Andersen, B., Bonefeld, C. M., Bzorek, M., Løvendorf, M. B., Lauritsen, J. P. H., Skov, L., and Geisler, C.
- Subjects
THERAPEUTIC use of vitamin D ,PSORIASIS treatment ,INTERLEUKIN-17 ,IMMUNOMODULATORS ,CD8 antigen ,IMMUNOLOGICAL adjuvants ,IMMUNOHISTOCHEMISTRY ,THERAPEUTICS - Abstract
The vitamin D analogue calcipotriol is an immunomodulatory drug widely used to treat psoriasis; however, how calcipotriol affects the immune cells in psoriasis lesions is not fully understood. The aim of this atudy was to investigate the effect of calcipotriol on the frequency of CD4
+ and CD8+ T cells and innate lymphoid cells ( ILC) and their production of IL-17A, IFN- γ and IL-22 in psoriasis lesions in patients with chronic plaque psoriasis. Eighteen patients with psoriasis were included, and two similar psoriasis lesions were chosen for each patient. One lesion was treated with calcipotriol (50 μg/g) and the other with vehicle twice a day for 14 days. The clinical effect was measured by degree of erythema, scaling and induration in each lesion ( SUM score). Skin biopsies were collected for histological and immunohistochemical analyses. Skin-derived cells were isolated and analysed by flow cytometry. After 14 days of treatment with calcipotriol, a significant clinical and histological effect was seen; however, we found no differences in the frequency of CD4+ and CD8+ T cells or ILC between calcipotriol- and vehicle-treated skin. The main finding was a significant decrease in CD8+ IL-17+ T cells in skin-derived cells from calcipotriol-treated skin, which was further supported by the absence of CD8+ IL-17+ T cells in immunohistochemical staining of calcipotriol-treated skin. No changes in the frequency of IL-22+ or IFN- γ+ cells were observed. Our findings show that the vitamin D analogue calcipotriol reduces the frequency of CD8+ IL-17+ T cells in psoriasis lesions concomitant with clinical improvement. [ABSTRACT FROM AUTHOR]- Published
- 2015
- Full Text
- View/download PDF
19. Correlation between Thymidylate SynthaseGene Variants, RNA and Protein Levels in Primary Colorectal Adenocarcinomas
- Author
-
Kristensen, MH, Weidinger, M, Bzorek, M, Pedersen, PL, and Mejer, J
- Abstract
This study was designed to compare thymidylate synthase (TS)genotype, mRNA and protein levels in primary colorectal adenocarcinoma, and to examine the correlation between microsatellite instability (MSI) and TSexpression. The TSgenotype of 68 patients with colorectal cancer was determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism analysis in peripheral blood mononuclear cells and tumour tissue. The TS mRNA levels in tumour tissue were measured by reverse-transcription PCR, and TS protein levels and MSI status were assessed using immunohistochemistry. Significantly higher mRNA and protein levels were observed in patients with the TS3R/3R versus the 2R/2R and 2R/3R genotypes. There was no correlation between TSsingle nucleotide polymorphism and TSexpression. Individuals homozygous for the six base-pair insertion in the 3′-untranslated region had significantly higher TS mRNA levels than heterozygous and homozygous wild typeindividuals. The TS mRNA and protein levels were significantly higher in microsatellite unstable tumours compared with microsatellite stable tumours. There was a significant association between the number of TSenhancer region repeats (in blood) and intratumoural TS mRNA and protein levels. A larger case series investigating the role of TSgene polymorphisms as predictors of sensitivity to 5-fluorouracil-based chemotherapy is required.
- Published
- 2010
- Full Text
- View/download PDF
20. Stage-related increase in PIM2 expression in mycosis fungoides.
- Author
-
Nielsen MH, Nielsen PR, Bzorek M, Eriksen JO, Wehkamp U, Lindahl LM, Woetmann A, Ødum N, Litman T, and Gjerdrum LMR
- Subjects
- Humans, Male, Female, Middle Aged, Adult, Aged, Immunohistochemistry, Skin Neoplasms genetics, Skin Neoplasms pathology, Aged, 80 and over, Biopsy, Skin pathology, Skin metabolism, Young Adult, Mycosis Fungoides genetics, Mycosis Fungoides pathology, Mycosis Fungoides metabolism, Proto-Oncogene Proteins genetics, Proto-Oncogene Proteins metabolism, Protein Serine-Threonine Kinases genetics, Protein Serine-Threonine Kinases metabolism
- Abstract
The oncogene PIM2 is upregulated in several malignancies but has never been investigated in mycosis fungoides (MF), the most common type of cutaneous T-cell lymphoma (CTCL). PIM2 is a well-known oncogene and is regulated by cell signaling pathways like the JAK/STAT- and NF-kB-pathway, key regulators in the pathogenesis of CTCL. The aim of this study was to examine the role of PIM2 in MF. PIM2 gene expression was measured in 81 formalin-fixed paraffin-embedded skin biopsies from patients with MF and 46 control biopsies from healthy skin (HS) and benign inflammatory skin disease (BID). Validation of PIM2 protein expression was performed on selected biopsies with immunohistochemical staining. We found a significant difference in gene expression levels between both early stage MF and HS (p < 0.0001), and BID (p < 0.0001). In addition, the PIM2 gene expression was higher in advanced-stage MF compared to early stage disease (p = 0.0001). No significant difference in gene expression levels was found between patients with and without disease progression. In conclusion, we found PIM2 expression is significantly increased in MF compared to controls, and in advanced-stage MF compared to early stage MF. These findings could potentially have diagnostic value in discriminating early stage MF from BID., (© 2024 Scandinavian Societies for Pathology, Medical Microbiology and Immunology.)
- Published
- 2024
- Full Text
- View/download PDF
21. Comparative Quantitative Proteomic Analysis of Melanoma Subtypes, Nevus-Associated Melanoma, and Corresponding Nevi.
- Author
-
Naimy S, Sølberg JBK, Kuczek DE, Løvendorf MB, Bzorek M, Litman T, Mund A, Rahbek Gjerdrum LM, Clark RA, Mann M, and Dyring-Andersen B
- Subjects
- Humans, Female, Male, Middle Aged, Aged, Proteome analysis, Proteome metabolism, Adult, Signal Transduction, Laser Capture Microdissection, Mass Spectrometry, Melanoma, Cutaneous Malignant, Melanoma pathology, Melanoma metabolism, Skin Neoplasms pathology, Skin Neoplasms metabolism, Proteomics methods, Nevus pathology, Nevus metabolism
- Abstract
A substantial part of cutaneous malignant melanomas develops from benign nevi. However, the precise molecular events driving the transformation from benign to malignant melanoma are not well-understood. We used laser microdissection and mass spectrometry to analyze the proteomes of melanoma subtypes, including superficial spreading melanomas (n = 17), nodular melanomas (n = 17), and acral melanomas (n = 15). Furthermore, we compared the proteomes of nevi cells with those of melanoma cells within the same specimens (nevus-associated melanoma (n = 14)). In total, we quantified 7935 proteins. Despite the genomic and clinical differences of the melanoma subtypes, our analysis revealed relatively similar proteomes, except for the upregulation of proteins involved in immune activation in nodular melanomas versus acral melanomas. Examining nevus-associated melanoma versus nevi, we found 1725 differentially expressed proteins (false discovery rate < 0.05). Among these proteins were 140 that overlapped with cancer hallmarks, tumor suppressors, and regulators of metabolism and cell cycle. Pathway analysis indicated aberrant activation of the phosphoinositide 3-kinase-protein kinase B-mTOR pathways and the Hippo-YAP pathway. Using a classifier, we identified six proteins capable of distinguishing melanoma from nevi samples. Our study represents a comprehensive comparative analysis of the proteome in melanoma subtypes and associated nevi, offering insights into the biological behavior of these distinct entities., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2024
- Full Text
- View/download PDF
22. Topical Neuropeptide Y for Ischemic Skin Wounds.
- Author
-
Stangerup T, Gjerdrum LMR, Bzorek M, Andersen L, Heegaard AM, Jorgensen LN, and Ågren MS
- Subjects
- Rats, Male, Animals, Rats, Sprague-Dawley, Receptors, Neuropeptide Y, Hydrogels pharmacology, Neuropeptide Y genetics, Neuropeptide Y pharmacology, Wound Healing
- Abstract
Our objective was to investigate the effects of topically applied neuropeptide Y (NPY) on ischemic wounds. Initially, the animal model for ischemic wound healing was validated using 16 male Sprague Dawley albino rats. In the intervention study, an additional 28 rats were divided into three groups: NPY (0.025%), the positive control insulin-like growth factor-I (IGF-I, 0.0025%), and the hydrogel carrier alone (control). The hydrogel was selected due to its capacity to prolong NPY release ( p < 0.001), as demonstrated in a Franz diffusion cell. In the animals, an 8 mm full-thickness wound was made in a pedunculated dorsal ischemic skin flap. Wounds were then treated and assessed for 14 days and collected at the end of the experiment for in situ hybridization analysis (RNAscope
® ) targeting NPY receptor Y2R and for meticulous histologic examination. Wound healing rates, specifically the percentage changes in wound area, did not show an increase with NPY ( p = 0.907), but there was an increase with rhIGF-I ( p = 0.039) compared to the control. Y2R mRNA was not detected in the wounds or adjacent skin but was identified in the rat brain (used as a positive control). Light microscopic examination revealed trends of increased angiogenesis and enhanced inflammatory cell infiltration with NPY compared to control. An interesting secondary discovery was the presence of melanophages in the wounds. Our findings suggest the potential of NPY to enhance neovascularization under ischemic wound healing conditions, but further optimization of the carrier and dosage is necessary. The mechanism remains elusive but likely involves NPY receptor subtypes other than Y2R.- Published
- 2024
- Full Text
- View/download PDF
23. Digitally assessed lymphocyte infiltration in rectal cancer biopsies is associated with pathological response to neoadjuvant therapy.
- Author
-
Jepsen DNM, Høeg H, Bzorek M, Orhan A, Eriksen JO, Gögenur I, Reiss B, and Fiehn AK
- Subjects
- Humans, Neoadjuvant Therapy methods, Chemoradiotherapy methods, Biomarkers, Biopsy, Rectal Neoplasms therapy, Rectal Neoplasms pathology, Carcinoma
- Abstract
A frequently used treatment strategy in locally advanced rectal cancer (RC) is neoadjuvant therapy followed by surgery. Patients treated with neoadjuvant therapy achieve varying pathological response, and currently, predicting the degree of response is challenging. This study examined the association between digitally assessed histopathological features in the diagnostic biopsies and pathological response to neoadjuvant therapy, aiming to find potential predictive biomarkers. 50 patients with RC treated with neoadjuvant chemotherapy and/or radiotherapy followed by surgery were included. Deep learning-based digital algorithms were used to assess the epithelium tumor area percentage (ETP) based on H&E-stained slides, and to quantify the density of CD3
+ and CD8+ lymphocytes, as well as the CD8+ /CD3+ lymphocyte percentage, based on immunohistochemically stained slides, from the diagnostic tumor biopsies. Pathological response was assessed according to the Mandard method. A good pathological response was defined as tumor regression grade (TRG) 1-2, and a complete pathological response was defined as Mandard TRG 1. Associations between the ETP and lymphocyte densities in the diagnostic biopsies and the pathological response were examined. The density of CD8+ lymphocytes, and the CD8+ /CD3+ lymphocyte percentage, were associated with both good and complete response to neoadjuvant therapy, while the density of CD3+ lymphocytes was associated with complete response. The ETP did not correlate with response to neoadjuvant therapy. It is well-known that infiltration of lymphocytes in colorectal cancer is a prognostic biomarker. However, assessment of CD8+ and CD3+ lymphocytes in the diagnostic tumor biopsies of patients with RC may also be useful in predicting response to neoadjuvant therapy., Competing Interests: Declaration of competing interest HH and BR are employed at Visiopharm, Hørsholm, A/S. The other authors report no conflict of interest related to the work described., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)- Published
- 2024
- Full Text
- View/download PDF
24. LAG3 and TIGIT Expression on Tumor-Infiltrating Lymphocytes in Cutaneous Melanoma.
- Author
-
Naimy S, Bzorek M, Eriksen JO, Løvendorf MB, Litman T, Dyring-Andersen B, and Gjerdrum LMR
- Subjects
- Humans, CD8-Positive T-Lymphocytes, Lymphocytes, Tumor-Infiltrating, Programmed Cell Death 1 Receptor genetics, Programmed Cell Death 1 Receptor metabolism, Receptors, Immunologic genetics, Receptors, Immunologic metabolism, Tumor Microenvironment, Melanoma pathology, Skin Neoplasms pathology
- Abstract
Background: Melanoma is widely recognized to be an immunogenic tumor that often contains tumor-infiltrating lymphocytes (TILs) in the tumor microenvironment. During cancer progression, expression of ligands that bind immune checkpoint (IC) proteins, such as PD-1, expressed on the surface of TILs, hinder them from exerting their antitumor functions. TILs consist of a heterogenous group of immune cells and their presence is associated with an improved overall survival in melanoma patients. Introduction of IC inhibitors has revolutionized management and prognosis of advanced melanoma. Unfortunately, the response rates have continued to be limited, resulting in growing interest in characterizing novel IC proteins, and developing combination therapy that includes inhibitors against multiple IC proteins., Methods: In a regional cohort of 166 patients diagnosed with cutaneous superficial spreading melanoma with different degree of TILs, we investigated the tumor immune-associated gene expression profile using NanoString Technology. We used multiplex immunofluorescence (mIF) staining in a subset of tumors (N = 7), combining IC proteins T-cell immunoglobulin and ITIM domain (TIGIT) and LAG3 with a melanoma cell marker (SOX10) and immune cell markers (CD8 [cytotoxic T cells], CD4 [T helper cells], FOXP3 [regulatory T cells/Tregs], PAX5 [B cells], and CD56 [NK/NKT cells]) and IC protein PD-1., Results: We found upregulation of 91 differentially expressed genes, including IC proteins, LAG3 and TIGIT in melanomas with brisk TILs compared to tumors where TILs were absent. mIF staining revealed LAG3 and TIGIT expression in the majority of CD8+ T cells. Only few Tregs and CD4+ T cells expressed LAG3, whereas majority of them expressed TIGIT. LAG3 and TIGIT were expressed in a small fraction of the NK/NKT cells and lacked in the B cells. The majority of PD-1+ cells co-localized with LAG3 and TIGIT., Conclusion: We report a variable expression of LAG3 and TIGIT on TILs subtypes and a coeval occurrence with PD-1. This knowledge places LAG3 and TIGIT in spatial and cellular context in melanoma. The data suggest that targeting multiple IC proteins might help overcome the current challenges with IC therapies., (© 2023 The Author(s). Published by S. Karger AG, Basel.)
- Published
- 2024
- Full Text
- View/download PDF
25. A Standardized and Reproducible Workflow for Membrane Glass Slides in Routine Histology and Spatial Proteomics.
- Author
-
Nordmann TM, Schweizer L, Metousis A, Thielert M, Rodriguez E, Rahbek-Gjerdrum LM, Stadler PC, Bzorek M, Mund A, Rosenberger FA, and Mann M
- Abstract
Defining the molecular phenotype of single cells in situ is key for understanding tissue architecture in health and disease. Advanced imaging platforms have recently been joined by spatial omics technologies, promising unparalleled insights into the molecular landscape of biological samples. Furthermore, high-precision laser microdissection (LMD) of tissue on membrane glass slides is a powerful method for spatial omics technologies and single-cell type spatial proteomics in particular. However, current histology protocols have not been compatible with glass membrane slides and LMD for automated staining platforms and routine histology procedures. This has prevented the combination of advanced staining procedures with LMD. In this study, we describe a novel method for handling glass membrane slides that enables automated eight-color multiplexed immunofluorescence staining and high-quality imaging followed by precise laser-guided extraction of single cells. The key advance is the glycerol-based modification of heat-induced epitope retrieval protocols, termed "G-HIER." We find that this altered antigen-retrieval solution prevents membrane distortion. Importantly, G-HIER is fully compatible with current antigen retrieval workflows and mass spectrometry-based proteomics and does not affect proteome depth or quality. To demonstrate the versatility of G-HIER for spatial proteomics, we apply the recently introduced deep visual proteomics technology to perform single-cell type analysis of adjacent suprabasal and basal keratinocytes of human skin. G-HIER overcomes previous incompatibility of standard and advanced staining protocols with membrane glass slides and enables robust integration with routine histology procedures, high-throughput multiplexed imaging, and sophisticated downstream spatial omics technologies., Competing Interests: Conflict of interest M. M. is an indirect investor in Evosep Biosciences. All other authors declare no competing interests., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
- Full Text
- View/download PDF
26. Lessons Learned, Challenges Taken, and Actions Made for "Precision" Immunohistochemistry. Analysis and Perspectives From the NordiQC Proficiency Testing Program.
- Author
-
Nielsen S, Bzorek M, Vyberg M, and Røge R
- Subjects
- Humans, Female, Reproducibility of Results, Immunohistochemistry, Quality Control, Receptors, Estrogen, Biomarkers, Tumor analysis, Receptor, ErbB-2, Laboratory Proficiency Testing methods, Breast Neoplasms diagnosis
- Abstract
Immunohistochemistry (IHC) has for decades been an integrated method within pathology applied to gain diagnostic, prognostic, and predictive information. However, the multimodality of the analytical phase of IHC is a challenge to ensure the reproducibility of IHC, which has been documented by external quality assessment (EQA) programs for many biomarkers. More than 600 laboratories participate in the Nordic immunohistochemical Quality Control EQA program for IHC. In the period, 2017-2021, 65 different biomarkers were assessed and a total of 31,967 results were evaluated. An overall pass rate of 79% was obtained being an improvement compared with 71% for the period, 2003-2015. The pass rates for established predictive biomarkers (estrogen receptor, progesterone receptor, and HER2) for breast carcinoma were most successful showing mean pass rates of 89% to 92%. Diagnostic IHC biomarkers as PAX8, SOX10, and different cytokeratins showed a wide spectrum of pass rates ranging from 37% to 95%, mean level of 75%, and attributed to central parameters as access to sensitive and specific antibodies but also related to purpose of the IHC test and validation performed accordingly to this. Seven new diagnostic biomarkers were introduced, and all showed inferior pass rates compared with the average level for diagnostic biomarkers emphasizing the challenge to optimize, validate, and implement new IHC biomarkers. Nordic immunohistochemical Quality Control operates by "Fit-For-Purpose" EQA principles and for programmed death-ligand 1, 2 segments are offered aligned to the "3-dimensional" approach-bridging diagnostic tests, drugs to be offered, and diseases addressed. Mean pass rates of 65% and 79% was obtained in the 2 segments for programmed death-ligand 1., Competing Interests: The authors declare no conflict of interest., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc.)
- Published
- 2023
- Full Text
- View/download PDF
27. Endoscopic calcium electroporation for colorectal cancer: a phase I study.
- Author
-
Broholm M, Vogelsang R, Bulut M, Stigaard T, Falk H, Frandsen S, Pedersen DL, Perner T, Fiehn AK, Mølholm I, Bzorek M, Rosen AW, Andersen CSA, Pallisgaard N, Gögenur I, and Gehl J
- Abstract
Background and study aims Colorectal cancer is one of the most common malignancies, with approximately 20 % of patients having metastatic disease. Local symptoms from the tumor remain a common issue and affect quality of life. Electroporation is a method to permeabilize cell membranes with high-voltage pulses, allowing increased passage of otherwise poorly permeating substances such as calcium. The aim of this study was to determine the safety of calcium electroporation for advanced colorectal cancer. Patients and methods Six patients with inoperable rectal and sigmoid colon cancer were included, all presenting with local symptoms. Patients were offered endoscopic calcium electroporation and were followed up with endoscopy and computed tomography/magnetic resonance scans. Biopsies and blood samples were collected at baseline and at follow-up, 4, 8, and 12 weeks after treatment. Biopsies were examined for histological changes and immunohistochemically with CD3/CD8 and PD-L1. In addition, blood samples were examined for circulating cell-free DNA (cfDNA). Results A total of 10 procedures were performed and no serious adverse events occurred. Prior to inclusion, patients reported local symptoms, such as bleeding (N = 3), pain (N = 2), and stenosis (N = 5). Five of six patients reported symptom relief. In one patient, also receiving systemic chemotherapy, clinical complete response of primary tumor was seen. Immunohistochemistry found no significant changes in CD3 /CD8 levels or cfDNA levels after treatment. Conclusions This first study of calcium electroporation for colorectal tumors shows that calcium electroporation is a safe and feasible treatment modality for colorectal cancer. It can be performed as an outpatient treatment and may potentially be of great value for fragile patients with limited treatment options., Competing Interests: Competing interests The authors declare that they have no conflict of interest., (The Author(s). This is an open access article published by Thieme under the terms of the Creative Commons Attribution-NonDerivative-NonCommercial License, permitting copying and reproduction so long as the original work is given appropriate credit. Contents may not be used for commercial purposes, or adapted, remixed, transformed or built upon. (https://creativecommons.org/licenses/by-nc-nd/4.0/).)
- Published
- 2023
- Full Text
- View/download PDF
28. Malignant T cells induce skin barrier defects through cytokine-mediated JAK/STAT signaling in cutaneous T-cell lymphoma.
- Author
-
Gluud M, Pallesen EMH, Buus TB, Gjerdrum LMR, Lindahl LM, Kamstrup MR, Bzorek M, Danielsen M, Bech R, Monteiro MN, Blümel E, Willerslev-Olsen A, Lykkebo-Valløe A, Vadivel CK, Krejsgaard T, Bonefeld CM, Geisler C, Becker JC, Koralov SB, Iversen L, Litman T, Woetmann A, and Ødum N
- Subjects
- Humans, Filaggrin Proteins, Quality of Life, T-Lymphocytes pathology, Cytokines metabolism, Lymphoma, T-Cell, Cutaneous pathology, Skin Diseases pathology, Skin Neoplasms pathology
- Abstract
Cutaneous T-cell lymphoma (CTCL) is a devastating lymphoid malignancy characterized by the accumulation of malignant T cells in the dermis and epidermis. Skin lesions cause serious symptoms that hamper quality of life and are entry sites for bacterial infection, a major cause of morbidity and mortality in advanced diseases. The mechanism driving the pathological processes that compromise the skin barrier remains unknown. Here, we report increased transepidermal water loss and compromised expression of the skin barrier proteins filaggrin and filaggrin-2 in areas adjacent to TOX-positive T cells in CTCL skin lesions. Malignant T cells secrete mediators (including cytokines such as interleukin 13 [IL-13], IL-22, and oncostatin M) that activate STAT3 signaling and downregulate filaggrin and filaggrin-2 expression in human keratinocytes and reconstructed human epithelium. Consequently, the repression of filaggrins can be counteracted by a cocktail of antibodies targeting these cytokines/receptors, small interfering RNA-mediated knockdown of JAK1/STAT3, and JAK1 inhibitors. Notably, we show that treatment with a clinically approved JAK inhibitor, tofacitinib, increases filaggrin expression in lesional skin from patients with mycosis fungoides. Taken together, these findings indicate that malignant T cells secrete cytokines that induce skin barrier defects via a JAK1/STAT3-dependent mechanism. As clinical grade JAK inhibitors largely abrogate the negative effect of malignant T cells on skin barrier proteins, our findings suggest that such inhibitors provide novel treatment options for patients with CTCL with advanced disease and a compromised skin barrier., (© 2023 by The American Society of Hematology.)
- Published
- 2023
- Full Text
- View/download PDF
29. BRAFV600E Expression Is Homogenous and Associated with Nonrecurrent Disease and Better Survival in Primary Melanoma.
- Author
-
Naimy S, Bzorek M, Eriksen JO, Dyring-Andersen B, and Rahbek Gjerdrum LM
- Subjects
- Humans, Proto-Oncogene Proteins B-raf genetics, Signal Transduction, Mutation, Melanoma, Cutaneous Malignant, Melanoma genetics, Melanoma pathology, Skin Neoplasms genetics, Skin Neoplasms pathology
- Abstract
Background: Superficial spreading melanomas (SSMs) are the most common type of melanoma and cause the majority of skin cancer deaths. More than 50% of cases harbor a mutation in the BRAF gene that activates the mitogen-activated protein kinase (MAPK) cancer signaling pathway. BRAFV600E is the most common BRAF mutation, and it represents an important biomarker that guides treatment selection. However, the relationship between the BRAFV600E gene expression and intratumoral protein distribution, on one side, and clinicopathological factors and patient outcomes, on the other, is not fully described. Additionally, whether MAPK cancer signaling activation in melanoma is due to increased biochemical activity of BRAFV600E, increased mRNA levels, or both requires further investigation. Here, we addressed these questions by examining expression patterns of BRAFV600E in primary treatment-naive melanomas and correlating them to clinicopathological factors and patient outcomes., Methods: In 166 SSM cases, we performed immunohistochemical staining to investigate the protein expression of BRAFV600E, and we measured BRAF mRNA levels using NanoString nCounter system., Results: Ninety-seven (49%) melanomas stained positive for BRAFV600E, with nearly 100% intratumoral homogeneity observed. Positive BRAFV600E expression was significantly associated with nonrecurrent disease and was found to be an independent predictor of better prognosis in univariate and multivariable analyses. Furthermore, presence of tumor-infiltrating lymphocytes, sentinel lymph node biopsy negativity, and low Breslow thickness were all independent predictors of better prognosis. We observed no difference in the BRAF mRNA levels in BRAFV600E-negative and BRAFV600E-positive melanomas, respectively. Validation in a larger publicly available cohort confirmed that there is only a weak correlation (Spearman 0.4) between BRAFV600E mRNA and protein levels and no differences in mRNA between BRAFV600E mutated and non-mutated patients., Conclusion: Our findings indicated that BRAFV600E is homogeneously present throughout the whole tumor and is associated with nonrecurrent disease and better survival in primary melanoma. We also showed that BRAFV600E mutation does not result in higher transcriptional levels, suggesting that activation of the MAPK signaling pathway in BRAFV600E mutated patients can be attributed to the increased biochemical activity caused by the mutation., (© 2023 The Author(s). Published by S. Karger AG, Basel.)
- Published
- 2023
- Full Text
- View/download PDF
30. Gene Expression Linked to Reepithelialization of Human Skin Wounds.
- Author
-
Ågren MS, Litman T, Eriksen JO, Schjerling P, Bzorek M, and Gjerdrum LMR
- Subjects
- Humans, Fibroblasts metabolism, Keratinocytes metabolism, RNA, Messenger genetics, Gene Expression, Skin injuries, Wounds and Injuries genetics
- Abstract
Our understanding of the regulatory processes of reepithelialization during wound healing is incomplete. In an attempt to map the genes involved in epidermal regeneration and differentiation, we measured gene expression in formalin-fixed, paraffin-embedded standardized epidermal wounds induced by the suction-blister technique with associated nonwounded skin using NanoString technology. The transcripts of 139 selected genes involved in clotting, immune response to tissue injury, signaling pathways, cell adhesion and proliferation, extracellular matrix remodeling, zinc transport and keratinocyte differentiation were evaluated. We identified 22 upregulated differentially expressed genes (DEGs) in descending order of fold change ( MMP1 , MMP3 , IL6 , CXCL8 , SERPINE1 , IL1B , PTGS2 , HBEGF , CXCL5 , CXCL2 , TIMP1 , CYR61 , CXCL1 , MMP12 , MMP9 , HGF , CTGF , ITGB3 , MT2A , FGF7 , COL4A1 and PLAUR ). The expression of the most upregulated gene, MMP1 , correlated strongly with MMP3 followed by IL6 and IL1B . rhIL-1β, but not rhIL-6, exposure of cultured normal human epidermal keratinocytes and normal human dermal fibroblasts increased both MMP1 mRNA and MMP-1 protein levels, as well as TIMP1 mRNA levels. The increased TIMP1 in wounds was validated by immunohistochemistry. The six downregulated DEGs ( COL7A1 , MMP28 , SLC39A2 , FLG1 , KRT10 and FLG2 ) were associated with epidermal maturation. KLK8 showed the strongest correlation with MKI67 mRNA levels and is a potential biomarker for keratinocyte proliferation. The observed gene expression changes correlate well with the current knowledge of physiological reepithelialization. Thus, the gene expression panel described in this paper could be used in patients with impaired healing to identify possible therapeutic targets.
- Published
- 2022
- Full Text
- View/download PDF
31. Concomitant Inhibition of FASN and SREBP Provides a Promising Therapy for CTCL.
- Author
-
Chi C, Harth L, Galera MR, Torrealba MP, Vadivel CK, Geisler C, Bonefeld CM, Nielsen PR, Bzorek M, Becker JC, Gjerdrum LMR, Ødum N, and Woetmann A
- Abstract
Cutaneous T cell lymphoma (CTCL) is a group of non-Hodgkin's primary cutaneous T cell lymphomas, with Mycosis Fungoides and Sézary syndrome (SS) being the two most common subtypes. Fatty acid synthase (FASN) is a crucial enzyme that catalyses the biosynthesis of fatty acids, which has been reported to play an oncogenic role in various malignancies but not in CTCL so far. Herein, we show that FASN is highly expressed in CTCL cell lines and in peripheral blood mononuclear cells (PBMCs) from CTCL patients, while it is not in PBMCs from healthy individuals. The inhibition of FASN in CTCL cell lines impairs cell viability, survival, and proliferation, but, interestingly, it also increases FASN expression. However, inhibiting sterol regulatory element binding protein (SREBP), a transcription factor that promotes the expression of FASN, partially reversed the upregulation of FASN induced by FASN inhibitors. Thus, the combination of FASN and SREBP inhibitors enhanced the effects on both CTCL cell lines and PBMCs from SS patients, where a valid inhibition on cell proliferation could be verified. Importantly, compared to non-malignant cells, primary malignant cells are more sensitive to the inhibition of FASN and SREBP, making the combination of FASN and SREBP inhibitors a promising novel therapeutic strategy in CTCL.
- Published
- 2022
- Full Text
- View/download PDF
32. Development of a Fully Automated Method to Obtain Reproducible Lymphocyte Counts in Patients With Colorectal Cancer.
- Author
-
Fiehn AK, Reiss B, Gögenur M, Bzorek M, and Gögenur I
- Subjects
- CD8-Positive T-Lymphocytes, Humans, Lymphocyte Count, Lymphocytes, Tumor-Infiltrating pathology, Artificial Intelligence, Colorectal Neoplasms diagnosis, Colorectal Neoplasms pathology
- Abstract
Colorectal cancer (CRC) is the third most common cancer worldwide. Although clinical outcome varies among patients diagnosed within the same TNM stage it is the cornerstone in treatment decisions as well as follow-up programmes. Tumor-infiltrating lymphocytes have added value when evaluating survival outcomes. The aim of this study was to develop a fully automated method for quantification of subsets of T lymphocytes in the invasive margin and central tumor in patients with CRC based on Deep Learning powered artificial intelligence. The study cohort consisted of 163 consecutive patients with a primary diagnosis of CRC followed by a surgical resection. Double-labeling immunohistochemical staining with cytokeratin in combination with CD3 or CD8, respectively, was performed on 1 representative slide from each patient. Visiopharm Quantitative Digital Pathology software was used to develop Application Protocol Packages for visualization of architectural details (background, normal epithelium, cancer epithelium, surrounding tissue), identification of central tumor and invasive margin as well as subsequent quantitative analysis of immune cells. Fully automated counts for CD3 and CD8 positive T cells were obtained in 93% and 92% of the cases, respectively. In the remaining cases, manual editing was required. In conclusion, the development of a fully automated method for counting CD3 + and CD8 + lymphocytes in a cohort of patients with CRC provided excellent results eliminating not only observer variability in lymphocyte counts but also in identifying the regions of interest for the quantitative analysis. Validation of the performance of the Application Protocol Packages including clinical correlation is needed., Competing Interests: B.R. is employee at Visiopharm. The remaining authors declare no conflict of interest., (Copyright © 2022 Wolters Kluwer Health, Inc. All rights reserved.)
- Published
- 2022
- Full Text
- View/download PDF
33. Deep Visual Proteomics defines single-cell identity and heterogeneity.
- Author
-
Mund A, Coscia F, Kriston A, Hollandi R, Kovács F, Brunner AD, Migh E, Schweizer L, Santos A, Bzorek M, Naimy S, Rahbek-Gjerdrum LM, Dyring-Andersen B, Bulkescher J, Lukas C, Eckert MA, Lengyel E, Gnann C, Lundberg E, Horvath P, and Mann M
- Subjects
- Humans, Laser Capture Microdissection methods, Mass Spectrometry methods, Proteome chemistry, Melanoma genetics, Proteomics methods
- Abstract
Despite the availabilty of imaging-based and mass-spectrometry-based methods for spatial proteomics, a key challenge remains connecting images with single-cell-resolution protein abundance measurements. Here, we introduce Deep Visual Proteomics (DVP), which combines artificial-intelligence-driven image analysis of cellular phenotypes with automated single-cell or single-nucleus laser microdissection and ultra-high-sensitivity mass spectrometry. DVP links protein abundance to complex cellular or subcellular phenotypes while preserving spatial context. By individually excising nuclei from cell culture, we classified distinct cell states with proteomic profiles defined by known and uncharacterized proteins. In an archived primary melanoma tissue, DVP identified spatially resolved proteome changes as normal melanocytes transition to fully invasive melanoma, revealing pathways that change in a spatial manner as cancer progresses, such as mRNA splicing dysregulation in metastatic vertical growth that coincides with reduced interferon signaling and antigen presentation. The ability of DVP to retain precise spatial proteomic information in the tissue context has implications for the molecular profiling of clinical samples., (© 2022. The Author(s).)
- Published
- 2022
- Full Text
- View/download PDF
34. An exploration of immunohistochemistry-based prognostic markers in patients undergoing curative resections for colon cancer.
- Author
-
Bennedsen ALB, Cai L, Hasselager RP, Özcan AA, Mohamed KB, Eriksen JO, Eiholm S, Bzorek M, Fiehn AK, Hviid TVF, and Gögenur I
- Subjects
- Aged, Aged, 80 and over, Disease-Free Survival, Female, Humans, Immunohistochemistry, Male, Middle Aged, Prognosis, Retrospective Studies, Biomarkers, Tumor metabolism, Colonic Neoplasms metabolism, Colonic Neoplasms mortality, Colonic Neoplasms pathology, Colonic Neoplasms surgery
- Abstract
Background: The immune system recognizes and destroys cancer cells. However, cancer cells develop mechanisms to avoid detection by expressing cell surface proteins. Specific tumour cell surface proteins (e.g. HLA-G, PD-L1, CDX2) either alone or in combination with the relative presence of immune cells (CD3 and CD8 positive T-cells) in the tumour tissue may describe the cancer cells' ability to escape eradication by the immune system. The aim was to investigate the prognostic value of immunohistochemical markers in patients with colon cancer., Methods: We conducted a retrospective study including patients diagnosed with pT3 and pT4 colon cancers. Immunohistochemical staining with HLA-G, PD-L1, CDX2, CD3, and CD8 was performed on tissue samples with representation of the invasive margin. PD-L1 expression in tumour cells and immune cells was reported conjointly. The expression of CD3 and CD8 was reported as a merged score based on the expression of both markers in the invasive margin and the tumour centre. Subsequently, a combined marker score was established based on all of the markers. Each marker added one point to the score when unfavourable immunohistochemical features was present, and the score was categorized as low, intermediate or high depending on the number of unfavourable stains. Hazard ratios for recurrence, disease-free survival and mortality were calculated., Results: We included 188 patients undergoing colon cancer resections in 2011-2012. The median follow-up was 41.7 months, during which 41 (21.8%) patients had recurrence and 74 (39.4%) died. In multivariable regression analysis positive HLA-G expression (HR = 3.37, 95%CI [1.64-6.93]) was associated with higher recurrence rates, while a preserved CDX2 expression (HR = 0.23, 95%CI [0.06-0.85]) was associated with a lower risk of recurrence. An intermediate or high combined marker score was associated with increased recurrence rates (HR = 20.53, 95%CI [2.68-157.32] and HR = 7.56, 95%CI [1.06-54.16], respectively). Neither high expression of PD-L1 nor high CD3-CD8 score was significantly associated with recurrence rates. Patients with a high CD3-CD8 score had a significantly longer DFS and OS., Conclusions: In tumour cells, expression of HLA-G and loss of CDX2 expression were associated with cancer recurrence. In addition, a combination of certain tumour tissue biomarkers was associated with colorectal cancer recurrence., (© 2022. The Author(s).)
- Published
- 2022
- Full Text
- View/download PDF
35. Staphylococcus aureus Induces Signal Transducer and Activator of Transcription 5‒Dependent miR-155 Expression in Cutaneous T-Cell Lymphoma.
- Author
-
Willerslev-Olsen A, Gjerdrum LMR, Lindahl LM, Buus TB, Pallesen EMH, Gluud M, Bzorek M, Nielsen BS, Kamstrup MR, Rittig AH, Bonefeld CM, Krejsgaard T, Geisler C, Koralov SB, Litman T, Becker JC, Woetmann A, Iversen L, and Odum N
- Subjects
- Anti-Bacterial Agents pharmacology, Cell Line, Tumor, Humans, Enterotoxins toxicity, Lymphoma, T-Cell, Cutaneous etiology, MicroRNAs physiology, STAT5 Transcription Factor physiology, Skin Neoplasms etiology, Staphylococcus aureus pathogenicity
- Abstract
Staphylococcal enterotoxins are believed to fuel disease activity in cutaneous T-cell lymphoma. Recent data support this by showing that antibiotics inhibit malignant T cells in skin lesions in mycosis fungoides and Sézary syndrome, the most common forms of cutaneous T-cell lymphoma. Yet, it remains incompletely characterized how staphylococcal enterotoxins fuel disease activity. In this study, we show that staphylococcal enterotoxins induce the expression of the oncogenic microRNA miR-155 in primary malignant T cells. Thus, staphylococcal enterotoxins and Staphyloccocus aureus isolates from lesional skin of patients induce miR-155 expression at least partly through the IL-2Rg‒Jak‒signal transducer and activator of transcription 5 pathway, and the effect is augmented by the presence of nonmalignant T cells. Importantly, mycosis fungoides lesions harbor S. aureus, express Y-phosphorylated signal transducer and activator of transcription 5, and display enhanced miR-155 expression, when compared with nonlesional and healthy skin. Preliminary data show that aggressive antibiotic therapy is associated with decreased Y-phosphorylated signal transducer and activator of transcription 5 and miR-155 expression in lesional skin in two patients with Sézary syndrome. In conclusion, we show that S. aureus and its enterotoxins induce enhanced expression of oncogenic miR-155, providing mechanistic insight into the role of S. aureus in cutaneous T-cell lymphoma. Our findings support that environmental stimuli such as bacteria can fuel disease progression in cutaneous T-cell lymphoma., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
36. NordiQC Assessments of Synaptophysin Immunoassays.
- Author
-
Vyberg M, Nielsen S, Bzorek M, and Røge R
- Subjects
- Humans, Immunoassay, Laboratory Proficiency Testing, Quality Control, Synaptophysin metabolism
- Abstract
This paper is number 8 in a series developed through a partnership between ISIMM and NordiQC with the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program., Competing Interests: The authors declare no conflict of interest., (Copyright © 2021 Wolters Kluwer Health, Inc. All rights reserved.)
- Published
- 2021
- Full Text
- View/download PDF
37. Changes in abdominal subcutaneous adipose tissue phenotype following menopause is associated with increased visceral fat mass.
- Author
-
Abildgaard J, Ploug T, Al-Saoudi E, Wagner T, Thomsen C, Ewertsen C, Bzorek M, Pedersen BK, Pedersen AT, and Lindegaard B
- Subjects
- Aging, Body Fat Distribution, Estrogen Receptor alpha genetics, Estrogen Receptor alpha metabolism, Female, Humans, Inflammation pathology, Insulin metabolism, Insulin Resistance, Intra-Abdominal Fat metabolism, Menopause, Metabolic Syndrome pathology, Middle Aged, Peroxisome Proliferator-Activated Receptors genetics, Peroxisome Proliferator-Activated Receptors metabolism, Phenotype, Subcutaneous Fat, Abdominal metabolism, Intra-Abdominal Fat pathology, Subcutaneous Fat, Abdominal pathology
- Abstract
Menopause is associated with a redistribution of adipose tissue towards central adiposity, known to cause insulin resistance. In this cross-sectional study of 33 women between 45 and 60 years, we assessed adipose tissue inflammation and morphology in subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) across menopause and related this to menopausal differences in adipose tissue distribution and insulin resistance. We collected paired SAT and VAT biopsies from all women and combined this with anthropometric measurements and estimated whole-body insulin sensitivity. We found that menopause was associated with changes in adipose tissue phenotype related to metabolic dysfunction. In SAT, postmenopausal women showed adipocyte hypertrophy, increased inflammation, hypoxia and fibrosis. The postmenopausal changes in SAT was associated with increased visceral fat accumulation. In VAT, menopause was associated with adipocyte hypertrophy, immune cell infiltration and fibrosis. The postmenopausal changes in VAT phenotype was associated with decreased insulin sensitivity. Based on these findings we suggest, that menopause is associated with changes in adipose tissue phenotype related to metabolic dysfunction in both SAT and VAT. Whereas increased SAT inflammation in the context of menopause is associated with VAT accumulation, VAT morphology is related to insulin resistance., (© 2021. The Author(s).)
- Published
- 2021
- Full Text
- View/download PDF
38. A novel approach for microRNA in situ hybridization using locked nucleic acid probes.
- Author
-
Paulsen IW, Bzorek M, Olsen J, Grum-Schwensen B, Troelsen JT, and Pedersen OB
- Subjects
- Female, Formaldehyde chemistry, Humans, Male, Nucleic Acid Probes genetics, Paraffin Embedding methods, Sensitivity and Specificity, Tissue Fixation methods, In Situ Hybridization methods, MicroRNAs genetics, Oligonucleotides genetics
- Abstract
Identification of target tissue microRNAs (miR) using in situ hybridization (ISH), with digoxigenin-labeled locked nucleic acid (LNA) probes, is influenced by preanalytic parameters. To determine the best retrieval method for common microRNAs, a multiblock composed of paraffin-embedded tonsil, cervix, placenta, and hyperplastic prostate tissue were included. Tissue were fixed in 10% formalin in a range of 5-144 hours (h). Cut sections (5 μm) from the multiblock were subjected to combinations of pretreatment procedures: variable periods of proteinase K (PK) digestion or Heat-induced microRNA Retrieval (HmiRR) using target retrieval solution (TRS) pH 6.1 or 9, with or without enzymatic treatment (pepsin). Results for the overall categories: TRS pH 9 versus PK; p = 2.9e-23, TRS pH 9 versus TRS pH 6.1; p = 1.1e-14, TRS pH 6.1 versus PK; p = 2.9e-03. A long fixation time, resulted in the best microRNA preservation and staining intensity (long vs. short: p = 3.5e-47, long vs. moderate: p = 1.6e-44, moderate vs. short: p = 4.3e-16), was enhanced using HmiRR TRS pH 9 with or without pepsin providing high sensitivity and specificity. These observations conflict with other ISH techniques (e.g., messenger ribonucleic acid), which typically require shorter fixation periods, and therefore, further studies are warranted.
- Published
- 2021
- Full Text
- View/download PDF
39. Diagnostic Two-Gene Classifier in Early-Stage Mycosis Fungoides: A Retrospective Multicenter Study.
- Author
-
Nielsen PR, Eriksen JO, Lindahl LM, Wehkamp U, Bzorek M, Andersen G, Woetmann A, Iversen L, Ødum N, Litman T, and Gjerdrum LMR
- Subjects
- Multicenter Studies as Topic, Mycosis Fungoides classification, Mycosis Fungoides genetics, Retrospective Studies, Mycosis Fungoides diagnosis
- Published
- 2021
- Full Text
- View/download PDF
40. Spatially and cell-type resolved quantitative proteomic atlas of healthy human skin.
- Author
-
Dyring-Andersen B, Løvendorf MB, Coscia F, Santos A, Møller LBP, Colaço AR, Niu L, Bzorek M, Doll S, Andersen JL, Clark RA, Skov L, Teunissen MBM, and Mann M
- Subjects
- Biomarkers metabolism, Cells, Cultured, Humans, Proteomics, Skin anatomy & histology, Skin immunology, Proteome metabolism, Skin cytology, Skin metabolism
- Abstract
Human skin provides both physical integrity and immunological protection from the external environment using functionally distinct layers, cell types and extracellular matrix. Despite its central role in human health and disease, the constituent proteins of skin have not been systematically characterized. Here, we combine advanced tissue dissection methods, flow cytometry and state-of-the-art proteomics to describe a spatially-resolved quantitative proteomic atlas of human skin. We quantify 10,701 proteins as a function of their spatial location and cellular origin. The resulting protein atlas and our initial data analyses demonstrate the value of proteomics for understanding cell-type diversity within the skin. We describe the quantitative distribution of structural proteins, known and previously undescribed proteins specific to cellular subsets and those with specialized immunological functions such as cytokines and chemokines. We anticipate that this proteomic atlas of human skin will become an essential community resource for basic and translational research ( https://skin.science/ ).
- Published
- 2020
- Full Text
- View/download PDF
41. Expression of the Voltage-Gated Potassium Channel Kv1.3 in Lesional Skin from Patients with Cutaneous T-Cell Lymphoma and Benign Dermatitis.
- Author
-
Hu T, Krejsgaard T, Nastasi C, Buus TB, Nansen A, Hald A, Spee P, Nielsen PR, Blümel E, Gluud M, Willerslev-Olsen A, Woetmann A, Bzorek M, Eriksen JO, Ødum N, and Rahbek Gjerdrum LM
- Subjects
- Adolescent, Adult, Aged, Aged, 80 and over, Biopsy, Cell Line, Tumor, Child, Dermatitis pathology, Female, Humans, Immunohistochemistry, Kv1.3 Potassium Channel antagonists & inhibitors, Lymphoma, T-Cell, Cutaneous pathology, Male, Middle Aged, Skin pathology, Skin Neoplasms pathology, Young Adult, Dermatitis metabolism, Kv1.3 Potassium Channel biosynthesis, Lymphoma, T-Cell, Cutaneous metabolism, Skin metabolism, Skin Neoplasms metabolism
- Abstract
Background: The voltage-gated potassium channel Kv1.3 (KCNA3) is expressed by effector memory T cells (TEM) and plays an important role in their activation and proliferation. Mycosis fungoides (MF), the most common subtype of cutaneous T-cell lymphoma (CTCL), was recently proposed to be a malignancy of skin-resident TEM. However, the expression of Kv1.3 in CTCL has not been investigated., Objectives: This study aims to examine the expression of Kv1.3 in situ and in vitro in CTCL., Methods: The expression of Kv1.3 was examined by immunohistochemistry in skin lesions from 38 patients with MF, 4 patients with Sézary syndrome (SS), and 27 patients with benign dermatosis. In 4 malignant T-cell lines of CTCL (Myla2059, PB2B, SeAx, and Mac2a) and a non-malignant T-cell line (MyLa1850), the expression of Kv1.3 was determined by flow cytometry. The proliferation of those cell lines treated with various concentrations of Kv1.3 inhibitor ShK was measured by 3H-thymdine incorporation., Results: Half of the MF patients (19/38) displayed partial Kv1.3 expression including 1 patient with moderate Kv1.3 positivity, while the other half (19/38) exhibited Kv1.3 negativity. An almost identical distribution was observed in patients with benign conditions, that is, 44.4% (12/27) were partially positive for Kv1.3 including 1 patient with moderate Kv1.3 positivity, while 55.6% (15/27) were Kv1.3 negative. In contrast, 3 in 4 SS patients displayed partial Kv1.3 positivity including 2 patients with weak staining and 1 with moderate staining, while 1 in 4 SS patients was Kv1.3 negative. In addition, all malignant T-cell lines, and a non-malignant T-cell line, displayed low Kv1.3 surface expression with a similar pattern. Whereas 2 cell lines (PB2B and Mac2a) were sensitive to Kv1.3 blockade, the other 2 (Myla2059 and SeAx) were completely resistant., Conclusions: We provide the first evidence of a heterogeneous Kv1.3 expression in situ in CTCL lesions., (© 2019 S. Karger AG, Basel.)
- Published
- 2020
- Full Text
- View/download PDF
42. Antibiotics inhibit tumor and disease activity in cutaneous T-cell lymphoma.
- Author
-
Lindahl LM, Willerslev-Olsen A, Gjerdrum LMR, Nielsen PR, Blümel E, Rittig AH, Celis P, Herpers B, Becker JC, Stausbøl-Grøn B, Wasik MA, Gluud M, Fredholm S, Buus TB, Johansen C, Nastasi C, Peiffer L, Kubat L, Bzorek M, Eriksen JO, Krejsgaard T, Bonefeld CM, Geisler C, Mustelin T, Langhoff E, Givskov M, Woetmann A, Kilian M, Litman T, Iversen L, and Odum N
- Subjects
- Aged, Cell Proliferation drug effects, Female, Humans, Lymphoma, T-Cell, Cutaneous metabolism, Lymphoma, T-Cell, Cutaneous pathology, Male, Middle Aged, Prospective Studies, STAT3 Transcription Factor metabolism, Signal Transduction drug effects, Skin Neoplasms metabolism, Skin Neoplasms pathology, T-Lymphocytes drug effects, T-Lymphocytes metabolism, T-Lymphocytes pathology, Anti-Bacterial Agents therapeutic use, Lymphoma, T-Cell, Cutaneous drug therapy, Skin Neoplasms drug therapy
- Abstract
It has been proposed that CD4 T-cell responses to Staphylococcus aureus (SA) can inadvertently enhance neoplastic progression in models of skin cancer and cutaneous T-cell lymphoma (CTCL). In this prospective study, we explored the effect of transient antibiotic treatment on tumor cells and disease activity in 8 patients with advanced-stage CTCL. All patients experienced significant decrease in clinical symptoms in response to aggressive, transient antibiotic treatment. In some patients, clinical improvements lasted for more than 8 months. In 6 of 8 patients, a malignant T-cell clone could be identified in lesional skin, and a significant decrease in the fraction of malignant T cells was observed following antibiotics but an otherwise unchanged treatment regimen. Immunohistochemistry, global messenger RNA expression, and cell-signaling pathway analysis indicated that transient aggressive antibiotic therapy was associated with decreased expression of interleukin-2 high-affinity receptors (CD25), STAT3 signaling, and cell proliferation in lesional skin. In conclusion, this study provides novel evidence suggesting that aggressive antibiotic treatment inhibits malignant T cells in lesional skin. Thus, we provide a novel rationale for treatment of SA in advanced CTCL.
- Published
- 2019
- Full Text
- View/download PDF
43. NordiQC Assessments of Chromogranin A Immunoassays.
- Author
-
Røge R, Kristoffersen HL, Bzorek M, Nielsen O, and Vyberg M
- Subjects
- Humans, Immunoassay, Antibodies, Monoclonal chemistry, Chromogranin A metabolism, Clinical Laboratory Techniques standards, Laboratory Proficiency Testing
- Abstract
This paper is number 6 in a series developed through a partnership between ISIMM and Nordic IHC Quality Control with the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.
- Published
- 2019
- Full Text
- View/download PDF
44. NordiQC Assessments of CD117 Immunoassays.
- Author
-
Røge R, Bzorek M, Nielsen O, and Vyberg M
- Subjects
- Denmark, Diagnostic Tests, Routine, Gastrointestinal Neoplasms diagnosis, Humans, Quality Assurance, Health Care, Quality Control, Gastrointestinal Neoplasms metabolism, Hematopoietic Stem Cells metabolism, Immunoassay methods, Laboratory Proficiency Testing methods, Proto-Oncogene Proteins c-kit metabolism
- Abstract
This paper is the number 5 in a series developed through a partnership between ISIMM and NordiQC for the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency-testing program.
- Published
- 2019
- Full Text
- View/download PDF
45. NordiQC Assessments of MSH6 Immunoassays.
- Author
-
Vyberg M, Røge R, Bzorek M, and Nielsen O
- Subjects
- DNA-Binding Proteins immunology, Humans, Immunoassay methods, Immunoassay standards, DNA-Binding Proteins metabolism, Quality Control
- Abstract
This paper is number 4 in a series developed through a partnership between ISIMM and Nordic immunohistochemical Quality Control for the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.
- Published
- 2018
- Full Text
- View/download PDF
46. NordiQC Assessments of PAX8 Immunoassays.
- Author
-
Røge R, Nielsen O, Bzorek M, Nielsen S, and Vyberg M
- Subjects
- Automation, Laboratory, Clinical Laboratory Techniques, Cross Reactions, Humans, Laboratory Proficiency Testing, Observer Variation, PAX8 Transcription Factor immunology, Quality Control, Immunoassay methods, Immunohistochemistry methods, PAX8 Transcription Factor metabolism
- Abstract
This paper is number 3 in a series developed through a partnership between ISIMM and NordiQC for the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.
- Published
- 2018
- Full Text
- View/download PDF
47. HLA class Ia and Ib molecules and FOXP3+ TILs in relation to the prognosis of malignant melanoma patients.
- Author
-
Melsted WN, Johansen LL, Lock-Andersen J, Behrendt N, Eriksen JO, Bzorek M, Scheike T, and Hviid TVF
- Subjects
- Adult, Aged, Cohort Studies, Female, Forkhead Transcription Factors genetics, Gene Expression Regulation, Neoplastic physiology, Genes, MHC Class I genetics, Humans, Male, Melanoma genetics, Melanoma metabolism, Middle Aged, Prognosis, Skin Neoplasms genetics, Skin Neoplasms metabolism, Forkhead Transcription Factors metabolism, Genes, MHC Class I physiology, Lymphocytes, Tumor-Infiltrating physiology, Melanoma pathology, Skin Neoplasms pathology
- Abstract
HLA class Ia (HLA-ABC) and HLA class Ib (HLA-E, -F and -G) molecules and FOXP3+ tumor-infiltrating lymphocytes (TILs) are often reported as relevant factors of tumor immune regulation. We investigated their expression as prognostic factors in 200 patients with primary cutaneous melanoma (PCM). In our cohort, patients with tumors showing upregulation of HLA-ABC molecules had significantly thicker tumors (32% vs 7%, P<0.001), frequent ulceration (20% vs 6%, P=0.007) and frequent nodular melanomas (20% vs 4%, P=0.001). Additionally, high expression of HLA-G in the tumor was a sign of bad prognosis for the patients, being associated with thick tumors (30% vs 12%, P=0.017), ulceration (24% vs 5%, P<0.001) and positive sentinel node (13% vs 6%, P=0.015). HLA-E, HLA-F and FOXP3+ TILs were not indicative of the prognosis in PCM. High HLA-ABC and HLA-G were associated with tumor aggressiveness and could be relevant predictive markers for effective immunotherapy of melanoma tumors., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
- View/download PDF
48. Endometrial immune markers are potential predictors of normal fertility and pregnancy after in vitro fertilization.
- Author
-
Kofod L, Lindhard A, Bzorek M, Eriksen JO, Larsen LG, and Hviid TVF
- Subjects
- Adult, Female, Fertilization in Vitro, Humans, Japan, Pregnancy Outcome, Prognosis, Prospective Studies, Tacrolimus therapeutic use, Th1-Th2 Balance, Killer Cells, Natural immunology, Pregnancy, Th1 Cells immunology
- Abstract
Problem: Elucidating immune mechanisms in the endometrium, which lead to the success of implantation and pregnancy, is important in reproductive medicine. Studies of immune cell abundance have shown conflicting results, and the expression and importance of HLA class Ib proteins in pre-implantation endometrium have not yet been investigated., Method of Study: The study population consisted of four subgroups: a hydrosalpinx, a salpingectomy, an unexplained infertility, and a fertile control group. Endometrial samples were collected during the implantation window. Immune markers (CD56
+ and CD16+ cells, FoxP3+ Tregs, HLA-G, HLA-F) were quantified in the samples. The outcome of the subsequent IVF treatment was recorded., Results: Increased CD56+ uNK cells and high HLA-G expression served as predictor for successful pregnancy outcome. HLA-F expression was positively correlated with uNK cells, being indirectly predictive for achieving pregnancy., Conclusion: Endometrial uNK cell abundance in the pre-implantation endometrium seems to be important for normal fertility and pregnancy success, and they may be used as clinical markers to predict implantation success in IVF., (© 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)- Published
- 2017
- Full Text
- View/download PDF
49. NordiQC Assessments of SOX10 Immunoassays.
- Author
-
Røge R, Nielsen S, Bzorek M, and Vyberg M
- Subjects
- Humans, Tissue Array Analysis, Immunoassay methods, SOXE Transcription Factors metabolism
- Abstract
This paper is number 1 in a series developed through a partnership between ISIMM and NordiQC for the purpose of reporting research assessing the performance characteristics of immunoassays in an external proficiency testing program.
- Published
- 2017
- Full Text
- View/download PDF
50. The Vitamin D Analogue Calcipotriol Reduces the Frequency of CD8+ IL-17+ T Cells in Psoriasis Lesions.
- Author
-
Dyring-Andersen B, Bonefeld CM, Bzorek M, Løvendorf MB, Lauritsen JP, Skov L, and Geisler C
- Subjects
- Adult, Aged, CD4-Positive T-Lymphocytes cytology, CD8-Positive T-Lymphocytes cytology, Calcitriol therapeutic use, Erythema drug therapy, Erythema immunology, Female, Humans, Interferon-gamma metabolism, Interleukin-17 metabolism, Interleukins metabolism, Lymphocyte Count, Male, Middle Aged, Psoriasis immunology, Young Adult, Interleukin-22, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Calcitriol analogs & derivatives, Dermatologic Agents therapeutic use, Psoriasis drug therapy
- Abstract
The vitamin D analogue calcipotriol is an immunomodulatory drug widely used to treat psoriasis; however, how calcipotriol affects the immune cells in psoriasis lesions is not fully understood. The aim of this study was to investigate the effect of calcipotriol on the frequency of CD4(+) and CD8(+) T cells and innate lymphoid cells (ILC) and their production of IL-17A, IFN-γ and IL-22 in psoriasis lesions in patients with chronic plaque psoriasis. Eighteen patients with psoriasis were included, and two similar psoriasis lesions were chosen for each patient. One lesion was treated with calcipotriol (50 μg/g) and the other with vehicle twice a day for 14 days. The clinical effect was measured by degree of erythema, scaling and induration in each lesion (SUM score). Skin biopsies were collected for histological and immunohistochemical analyses. Skin-derived cells were isolated and analysed by flow cytometry. After 14 days of treatment with calcipotriol, a significant clinical and histological effect was seen; however, we found no differences in the frequency of CD4(+) and CD8(+) T cells or ILC between calcipotriol- and vehicle-treated skin. The main finding was a significant decrease in CD8(+) IL-17(+) T cells in skin-derived cells from calcipotriol-treated skin, which was further supported by the absence of CD8(+) IL-17(+) T cells in immunohistochemical staining of calcipotriol-treated skin. No changes in the frequency of IL-22(+) or IFN-γ(+) cells were observed. Our findings show that the vitamin D analogue calcipotriol reduces the frequency of CD8(+) IL-17(+) T cells in psoriasis lesions concomitant with clinical improvement., (© 2015 John Wiley & Sons Ltd.)
- Published
- 2015
- Full Text
- View/download PDF
Catalog
Discovery Service for Jio Institute Digital Library
For full access to our library's resources, please sign in.