Browsing by Subject " pathology"

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  • Syrjä, Pernilla; Anwar, Tahira; Pääkkönen, Tarja; Kyöstilä, Kaisa; Hultin Jäderlund, Karin; Cozzi, Francesca; Rohdin, Cecilia; Hahn, Kerstin; Wohlsein, Peter; Baumgärtner, Wolfgang; Henke, Diana; Oevermann, Anna; Sukura, Antti; Leeb, Tosso; Lohi, Hannes; Eskelinen, Eeva-Liisa (2017)
    A missense variant in the autophagy-related ATG4D-gene has been associated with a progressive degenerative neurological disease in Lagotto Romagnolo (LR) dogs. In addition to neural lesions, affected dogs show an extraneural histopathological phenotype characterized by severe cytoplasmic vacuolization, a finding not previously linked with disturbed autophagy in animals. Here we aimed at testing the hypothesis that autophagy is altered in the affected dogs, at reporting the histopathology of extraneural tissues and at excluding lysosomal storage diseases. Basal and starvation-induced autophagy were monitored by Western blotting and immunofluorescence of microtubule associated protein 1A/B light chain3 (LC3) in fibroblasts from 2 affected dogs. The extraneural findings of 9 euthanized LRs and skin biopsies from 4 living affected LRs were examined by light microscopy, electron microscopy, and immunohistochemistry (IHC), using antibodies against autophagosomal membranes (LC3), autophagic cargo (p62), and lysosomal membranes (LAMP2). Biochemical screening of urine and fibroblasts of 2 affected dogs was performed. Under basal conditions, the affected fibroblasts contained significantly more LC3-II and LC3-positive vesicles than did the controls. Morphologically, several cells, including serous secretory epithelium, endothelial cells, pericytes, plasma cells, and macrophages, contained cytoplasmic vacuoles with an ultrastructure resembling enlarged amphisomes, endosomes, or multivesicular bodies. IHC showed strong membranous LAMP2 positivity only in sweat glands. The results show that basal but not induced autophagy is altered in affected fibroblasts. The ultrastructure of affected cells is compatible with altered autophagic and endo-lysosomal vesicular traffic. The findings in this spontaneous disease provide insight into possible tissue-specific roles of basal autophagy.
  • Fagerstedt, K.W.; Salonen, T.; Zhao, F.; Kytölä, S.; Böhling, T.; Andersson, L.C. (2018)
    Myxoinflammatory fibroblastic sarcoma is a soft-tissue neoplasm most frequently found in the distal extremities of middle-aged adults. Most myxoinflammatory fibroblastic sarcoma are low-grade tumors with propensity for local recurrence after incomplete removal. We report a myxoinflammatory fibroblastic sarcoma which developed in the foot of a 41-year-old male and showed an exceptionally aggressive course with metastatic spread and fatal outcome within 16 months. We managed to establish a spontaneously transformed continuous cell line, called JU-PI, from a metastatic lesion. The JU-PI cells have a sub-tetraploid karyotype including the 1;10 chromosomal translocation and amplification of the proximal end of 3p; these features are considered genetic signatures of myxoinflammatory fibroblastic sarcoma. Both the primary tumor and the JU-PI cells showed nuclear expression of the TFE3 transcription factor but TFE3-activating chromosomal rearrangements were not found. To our knowledge, JU-PI is the first established myxoinflammatory fibroblastic sarcoma cell line. JU-PI cells offer a tool for investigating the molecular oncology of myxoinflammatory fibroblastic sarcoma. © 2018, © The Author(s) 2018.
  • Yates, Abi G.; Pink, Ryan C.; Erdbrugger, Uta; Siljander, Pia R-M.; Dellar, Elizabeth R.; Pantazi, Paschalia; Akbar, Naveed; Cooke, William R.; Vatish, Manu; Dias-Neto, Emmanuel; Anthony, Daniel C.; Couch, Yvonne (2022)
    It is clear from Part I of this series that extracellular vesicles (EVs) play a critical role in maintaining the homeostasis of most, if not all, normal physiological systems. However, the majority of our knowledge about EV signalling has come from studying them in disease. Indeed, EVs have consistently been associated with propagating disease pathophysiology. The analysis of EVs in biofluids, obtained in the clinic, has been an essential of the work to improve our understanding of their role in disease. However, to interfere with EV signalling for therapeutic gain, a more fundamental understanding of the mechanisms by which they contribute to pathogenic processes is required. Only by discovering how the EV populations in different biofluids change-size, number, and physicochemical composition-in clinical samples, may we then begin to unravel their functional roles in translational models in vitro and in vivo, which can then feedback to the clinic. In Part II of this review series, the functional role of EVs in pathology and disease will be discussed, with a focus on in vivo evidence and their potential to be used as both biomarkers and points of therapeutic intervention.
  • Laitinen, A.; Hagström, J.; Mustonen, H.; Kokkola, A.; Tervahartiala, T.; Sorsa, T.; Böckelman, C.; Haglund, C. (2018)
    Despite gastric cancer being rare nowadays in Western countries, it remains one of the leading causes of cancer death worldwide. The course of the disease varies, so the individual gastric cancer patient’s prognosis is difficult to determine. The need for new biomarkers is crucial. The aim of this study was to evaluate the prognostic value of serum matrix metalloproteinase-8, serum tissue inhibitor of metalloproteinase-1, and tissue matrix metalloproteinase-8 in patients with gastric cancer. Preoperative serum samples from 233 patients with gastric cancer were retrospectively analyzed. Serum levels of matrix metalloproteinase-8 were analyzed with immunofluorometric assay, and tissue inhibitor of metalloproteinase-1 levels were determined by enzyme-linked immunosorbent assay. We also determined the tissue expression of matrix metalloproteinase-8 in 276 gastric cancer samples by immunohistochemistry. Survival data and death causes came from patient records, the Population Register Center of Finland, and Statistics Finland. Patients with a low (131 ng/mL) serum matrix metalloproteinase-8 level had a considerably unfavorable prognosis (p = 0.002). Those patients with a high (≥170 ng/mL) serum tissue inhibitor of metalloproteinase-1 level also had a poor prognosis (p <0.001), and the latter remained significant in multivariable analysis (hazard ratio = 1.85; 95% confidence interval: 1.26–2.72; p = 0.002). The molar ratio of serum matrix metalloproteinase-8 and tissue inhibitor of metalloproteinase-1 levels with low (0.30) molar ratios predicted a worse prognosis (p = 0.020). Tissue matrix metalloproteinase-8 did not influence prognosis. These results suggest that serum matrix metalloproteinase-8, tissue inhibitor of metalloproteinase-1, and the ratio of matrix metalloproteinase-8/ tissue inhibitor of metalloproteinase-1 may prove useful biomarkers for prediction of prognosis in patients with gastric cancer. © The Author(s) 2018.
  • Lehtonen, Antti (Helsingin yliopisto, 2021)
    Soft tissue sarcoma (STS) is a rare neoplasm consisting of approximately 50 different histologic subtypes treated similarly. The median patient age is 60 years, and there’s a slight male predominance. There are no known predisposing factors apart from ionizing radiation. STS is diagnosed with core needle biopsy of a suspicious lump, most commonly a palpable, painless mass. Surgery with adequate margins is the only curative treatment of STS. Adjuvant radiation therapy (RT) improves local control rates (LC) and is offered for patients with positive surgical margins. However, there is no consensus on which surgical margin is wide enough. Adjuvant chemotherapy yields rather poor survival benefit with significant risk of toxicity and is therefore offered for minority of the patients. Prognostic factors for local recurrence include patient age, tumor grade and surgical margin. Due to disease rarity, complex diagnostics, need for surgical treatment, adjuvant radiation therapy and critical evaluation of patient selection to receive chemotherapy, centralization of treatment is effective. Specialist centers adhere to treatment guidelines more strictly than non-tertiary centers. Adequate preoperative imaging methods are more often used, adequate surgical margins are more often achieved, patients undergo fewer operations, adequate adjuvant RT is more often offered and local control rates are better in specialist centers compared to non-tertiary centers. However, there is still a lack of adherence to referral policies with only 63 percent of STSs referred before surgical intervention. The aim of this study was to quality control treatment received in Soft Tissue Sarcoma Group at Helsinki University Hospital (HUH) during its first 25 years, with special interest in 1) adherence to treatment protocol, 2) metastases-free survival (MFS) and overall survival (OS) of STS patients and 3) treatment and survival of radiation-associated STS. We used a 1327-patient series from 1987-2012 consisting of patients referred to STS Group at HUH. Patients were treated according to treatment protocol based on 1987 Scandinavian Sarcoma Group recommendations. 1182 (79 %) patients had only primary tumor at presentation. Only 411 (35 %) patients referred for a primary tumor were referred untouched. However, the proportion increased by time from 13 to 47 percent. Of the 1115 patients treated with a curative intent, 680 (61 %) patients underwent only one procedure, proportion of which increased by time. Only 64 and 62 percent of patients with intralesional (tumor tissue in resection margin) and marginal margins (smallest margin under 2.5 cm) received adjuvant RT. Overall survival rates for patients treated with curative intent were 68 percent and 55 percent in 5-year and 10-year follow-up, respectively. No improvement in OS was recorded. 59 patients were referred for a radiation-associated sarcoma, and 52 of them were treated with a curative intent. Tumor appeared in the radiation field of an invasive breast cancer in 77 percent of the patients. Local control rates for patients treated with curative intent were 73 percent and 58 percent in 5-year and 10-year follow-up, respectively. Adherence to treatment protocol in STS group at HUH has improved over time in 1987-2012. To increase the percentage of patients referred for primary tumor without preceding biopsy, increasing soft tissue sarcoma knowledge in district hospitals is vital. All exceptions from treatment protocol should be discussed and reported in patient files.
  • Fang, Shentong (Helsingin yliopisto, 2014)
    Most tissues possess tissue-specific stem cells that allow them to maintain tissue integrity. Stem cell niches provide an ideal regulatory microenvironment to support the maintenance and proliferation of adult stem cells. However, adult stem cells and stem cell niches have not been identified for all tissues. Adult stem cells that give rise to the vascular endothelium are still unknown. In this thesis work, we identified a rare population of vascular endothelial stem cells (VESC) on the vascular wall by the phenotype lin-CD31+CD105+Sca-1+c-kit+. A single c-kit expressing VESC with highly proliferative capacity generated functional blood vessels in vivo. A genetic defect in endothelial c-kit resulted in an abolished colony-forming ability of VESCs and impaired tumor angiogenesis and tumor growth. Angiogenesis, the growth of new blood vessels from pre-existing vessels, is actively involved in many physiological and pathological processes such as wound repair, female reproductive cycling, ischemic disease and tumor development. There are two major groups of cells involved in the adult vascular growth, cells that contribute directly by composing the blood vessels, such as vascular endothelial stem cells, and cells that contribute indirectly in a paracrine manner such as infiltrating hematopoietic cells. Infiltrating hematopoietic cells from the bone marrow contribute to angiogenesis in a paracrine manner by secreting angiogenic factors or by remodeling the extracellular matrix. In this thesis work, we found that transforming growth factor-β (TGF-β) recruited a massive amount of hematopoietic cells to local microenvironment. TGF-β stimulated vascular endothelial growth factor (VEGF) expression on these hematopoietic effectors and thus induced vascular growth. This stimulation was regulated by p38 and p44/p42 mitogen-activated protein kinase (MAPK) signaling pathways. These results together provided evidence for a dual action mechanism for TGF-β-induced angiogenesis in vivo. In malignant tumors, we found that the tumor expressed osteoblastic and vascular hematopoietic stem cell (HSC) niche molecules and enclosed multipotent hematopoietic progenitors. The proliferating hematopoietic progenitors generated hematopoietic effector cells and supported angiogenesis and tumor growth by secreting matrix metalloprotease 9 (MMP-9) and VEGF. HSPCs were found to be in proximity to tumor vasculature. Tumor microenvironment shared features of HSC niche in the bone marrow. Therapeutic ablation of hematopoietic cells including proliferating hematopoietic cells from tumor using AMD3100 in vivo resulted in inhibited tumor angiogenesis and growth. In conclusion, we identified and characterized a rare population of c-kit expressing VESCs that give rise to the vascular endothelium in adult. Further purification and detailed characterization of VESCs will provide a better understanding of VESCs and hierarchy of endothelial lineage. Our observations on the in vivo angiogenesis induced by TGF-β elucidated the mechanisms of action of TGF-β in promoting vascular growth. Together with current findings on the ectopic tumor hematopoietic microenvironment, the proliferating hematopoietic progenitors and angiogenic hematopoietic effector cells, these results provided potential therapeutic targets to treat cancer and other diseases.
  • Al-Rashed, F.; Ahmad, Z.; Iskandar, M.A.; Tuomilehto, J.; Al-Mulla, F.; Ahmad, R. (2019)
    Background/Aims: TNF-α-mediated pro-inflammatory phenotypic change in monocytes is known to be implicated in the pathogenesis of metabolic inflammation and insulin resistance. However, the mechanism by which TNF-α-induces inflammatory phenotypic shift in monocytes is poorly understood. Since long-chain acyl-CoA synthetase 1 (ACSL1) is associated with inflammatory monocytes/macrophages, we investigated the role of ACSL1 in the TNF-α-driven inflammatory phenotypic shift in the monocytes. Methods: Monocytes (Human monocytic THP-1 cells) were stimulated with TNF-α. Inflammatory phenotypic markers (CD16, CD11b, CD11c and HLA-DR) expression was determined with real time RT-PCR and flow cytometry. IL-1β and MCP-1 were determined by ELISA. Signaling pathways were identified by using ACSL1 inhibitor, ACSL1 siRNA and NF-κB reporter monocytic cells. Phosphorylation of NF-κB was analyzed by western blotting and flow cytometry. Results: Our data show that TNF-α induced significant increase in the expression of CD16, CD11b, CD11c and HLA-DR. Inhibition of ACSL1 activity in the cells with triacsin C significantly suppressed the expression of these inflammatory markers. Using ACSL-1 siRNA, we further demonstrate that TNF-α-induced inflammatory markers expression in monocytic cells requires ACSL1. In addition, IL-1b and MCP-1 production by TNF-α activated monocytic cells was significantly blocked by the inhibition of ACSL-1 activity. Interestingly, elevated NF-κB activity resulting from TNF-α stimulation was attenuated in ACSL1 deficient cells. Conclusion: Our findings provide an evidence that TNF-α-associated inflammatory polarization in monocytes is an ACSL1 dependent process, which indicates its central role in TNF-α-driven metabolic inflammation. © 2019 The Author(s).
  • Yin, Miao (Helsingin yliopisto, 2013)
    Within cancer, malignant cancer cells extensively interact with stromal components, which mainly consist of extracellular matrix (ECM) and several kinds of stromal cells to acquire the ability to proliferate without control, invade surrounding tissue, and metastasize to distant sites. During tumor development and progression, local invasion is a key step for tumor cells to disseminate to sentinel lymph nodes and distant organs. The aim of the present studies was to elucidate the interaction between melanoma cells and stromal components and identify molecules associated with melanoma and fibrosarcoma invasion. We have analyzed fibrosarcoma cells, which form highly invasive fibrosarcomas in nude mice. We found thymosin β4 (Tβ4) as the gene with the highest upregulation. Interestingly, a sponge toxin latrunculin A, which inhibits the binding of thymosin β4 to actin, was found to profoundly affect the morphology and proliferation of the fibrosarcoma cells and to block their invasion in three-dimensional Matrigel. In addition, our analyses revealed highly significant changes in genes encoding ECM proteins and ECM modifiers, such as versican, tenascin-C (TNC), thrombospondin 1 (TSP-1), cathepsin L(CTSL), and osteopontin (OPN). In the second study, melanoma cells and skin fibroblasts were cultured alone or together in three-dimensional (3D) Matrigel/collagen I gel models, mimicking the in vivo microenvironment to study the effects of skin fibroblasts on melanoma cell invasion. We compared the protease gene expression profiles of benign nevi, thin melanomas (< 2mm), and thick melanomas (≥ 2mm). Various protease inhibitors were then tested using 3D Matrigel invasion assay models. We also examined signaling involved in melanoma cell invasion. We found that the invasion of melanoma cells is promoted and guided by fibroblasts, and that this process is dependent on TGF-β signaling and cysteine cathepsins B/L, but not on MMP activity. Further, we demonstrated significantly increased expression of cathepsins B and L in invasive melanomas. Cathepsin B was predominantly expressed by the melanoma cells and cathepsin L by the fibroblasts. The expression level of cathepsin B was further found to correlate with shortened patient survival. In the third study, we have explored the role of OPN in promoting the malignant phenotype of melanoma cells cultured in 3D collagen-I gel matrix and the potential mechanisms. Invasive growth of primary melanoma cells overexpressing OPN was significantly stimulated, while apoptosis of OPN transfectants was clearly inhibited. Conversely, knockdown of OPN expression in metastatic melanoma cells caused growth and motility inhibition. Furthermore, we identified inverse expression of OPN and CD9 in melanocytes and melanoma cells.
  • Krebs, Rainer (Helsingin yliopisto, 2014)
    The aim of this study was to investigate the role of VEGF ligands A and C and their receptors (VEGFR-1, VEGFR-2, VEGFR-3) in the development of experimental OB. To this effect, tracheal allografts were transplanted heterotopically into MHC-mismatched mice and rats. In the heterotopic tracheal allograft model, an early inflammatory phase with its peak at 10 days after transplantation is followed by a fibroproliferative phase that results in complete luminal occlusion of the graft at 30 days after transplantation. We found that the expression of VEGF-A and VEGF-C as well as their receptors changed location and intensity during the development of experimental OB in tracheal allografts. The alloimmune response in tracheal transplants upregulated the expression of VEGF-A and VEGF-C primarily in graft-infiltrating mononuclear inflammatory cells and the microvasculature in the airway wall. The VEGF receptors showed a similar expression pattern. Overexpression of VEGF-A and VEGF-C significantly increased luminal occlusion at 30 days after transplantation. Blocking signaling through receptors VEGFR-1, VEGFR-2, and VEGFR-3, on the other hand, had beneficial effects and prevented the development of experimental OB. Blocking VEGFR-1 and VEGFR-2 alleviated the innate immune responses at three days after transplantation, especially on the natural killer cell level, thereby downscaling the subsequent adaptive immune response by CD4+ T cells at 10 days after transplantation and the development of luminal occlusion at 30 days after transplantation. Blocking VEGF-C/VEGFR-3 signaling inhibited intragraft lymphangiogenesis at both time points, influx of CD4+ T cells at 10 days, and traffic of dendritic cells out of the graft at 30 days after transplantation. These findings suggest a regulatory role for VEGF ligands in the pathogenesis of OB. Intervention at the VEGFR level could reduce early inflammatory reactions (VEGFR-1, VEGFR-2) or downscale antigen export and thus antigen presentation to alloreactive lymphocytes (VEGFR-3), thereby improving the long-term results of lung transplantation.
  • Geisslitz, Sabrina; Shewry, Peter; Brouns, Fred; America, Antoine H. P.; Caio, Giacomo Pietro Ismaele; Daly, Matthew; D'Amico, Stefano; De Giorgio, Roberto; Gilissen, Luud; Grausgruber, Heinrich; Huang, Xin; Jonkers, Daisy; Keszthelyi, Daniel; Larre, Colette; Masci, Stefania; Mills, Clare; Moller, Marie Sofie; Sorrells, Mark E.; Svensson, Birte; Zevallos, Victor F.; Weegels, Peter Louis (2021)
    Amylase/trypsin-inhibitors (ATIs) comprise about 2-4% of the total wheat grain proteins and may contribute to natural defense against pests and pathogens. However, they are currently among the most widely studied wheat components because of their proposed role in adverse reactions to wheat consumption in humans. ATIs have long been known to contribute to IgE-mediated allergy (notably Bakers' asthma), but interest has increased since 2012 when they were shown to be able to trigger the innate immune system, with attention focused on their role in coeliac disease which affects about 1% of the population and, more recently, in non-coeliac wheat sensitivity which may affect up to 10% of the population. This has led to studies of their structure, inhibitory properties, genetics, control of expression, behavior during processing, effects on human adverse reactions to wheat and, most recently, strategies to modify their expression in the plant using gene editing. We therefore present an integrated account of this range of research, identifying inconsistencies, and gaps in our knowledge and identifying future research needs. Note This paper is the outcome of an invited international ATI expert meeting held in Amsterdam, February 3-5 2020