Identification of Basophils as a Major Source of Hepatocyte Growth Factor in Chronic Myeloid Leukemia: A Novel Mechanism of BCR-ABL1-Independent Disease Progression

AbstractChronic myeloid leukemia (CML) is a hematopoietic neoplasm characterized by the Philadelphia chromosome and the related BCR-ABL1 oncoprotein. Acceleration of CML is usually accompanied by basophilia. Several proangiogenic molecules have been implicated in disease acceleration, including the hepatocyte growth factor (HGF). However, little is known so far about the cellular distribution and function of HGF in CML. We here report that HGF is expressed abundantly in purified CML basophils and in the basophil-committed CML line KU812, whereas all other cell types examined expressed only trace amounts of HGF or no HGF. Interleukin 3, a major regulator of human basophils, was found to promote HGF expression in CML basophils. By contrast, BCR-ABL1 failed to induce HGF synthesis in CML cells, and imatinib failed to inhibit expression of HGF in these cells. Recombinant HGF as well as basophil-derived HGF induced endothelial cell migration in a scratch wound assay, and these effects of HGF were reverted by an anti-HGF antibody as well as by pharmacologic c-Met inhibitors. In addition, anti-HGF and c-Met inhibitors were found to suppress the spontaneous growth of KU812 cells, suggesting autocrine growth regulation. Together, HGF is a BCR-ABL1-independent angiogenic and autocrine growth regulator in CML. Basophils are a unique source of HGF in these patients and may play a more active role in disease-associated angiogenesis and disease progression than has so far been assumed. Our data also suggest that HGF and c-Met are potential therapeutic targets in CML

18F-Fluorodeoxyglucose Uptake Level-Based Lymph Node Staging in Oropharyngeal Squamous Cell Cancer - Role of Molecular Marker Expression on Diagnostic Outcome

Background: A prospective study was performed to assess standard uptake value (SUV)-level based 18F-fluorodeoxyglucose (FDG)-positron emission tomography (PET)/computed tomography (CT) lymph node staging in 33 patients with oropharyngeal squamous cell cancer (OSCC) out of a total of 99 patients with head-and-neck squamous cell cancer (HNSCC) and the role of nodal molecular marker expression in diagnostic outcome prediction. Methods: Preoperative nodal PET/CT staging in 123 lymph nodes was correlated with postoperative lymph node histology, which served as gold standard. Tissue samples were prepared for immunohistochemistry of the excised lymph nodes. Results: The negative and positive predictive values (NPV and PPV) of PET for correct lymph node assessment were 100% and 93%, respectively. There was a significant association between SUVmax and lymph node histology (p < 0.0001) and a significant linear correlation between SUVmax and nodal size (Pearson’s correlation coefficient r = 0.61336, p < 0.0001). The molecular marker E-Cadherin was significantly overexpressed in lymph node metastases (p < 0.0001). Benign lymph nodes showed significant 2-fold Bcl2 overexpression (p < 0.0001). However, the molecular marker expression profiles were inhomogeneous and did not allow valuable diagnostic outcome prediction. Conclusions: SUV level-based 18F-FDG-PET/CT lymph node assessment in OSCC still has to be considered as the most established and reliable staging tool. Lymph node molecular marker expression profiles need to be investigated further as they currently do not sufficiently contribute to therapy decision-making

Clinical impact and proposed application of molecular markers, genetic variants, and cytogenetic analysis in mast cell neoplasms: Status 2022

Mast cell neoplasms are an emerging challenge in the fields of internal medicine, allergy, immunology, dermatology, laboratory medicine, and pathology. In this review, we discuss the current standards for the diagnosis and prognostication of mast cell neoplasms with special reference to clinically relevant germline and somatic gene variants. In patients with cutaneous mastocytosis or with indolent systemic mastocytosis (SM), various KIT-activating mutations act as key molecular drivers of the disease. In adults, KIT p.D816V is by far the most prevalent driver, whereas other KIT mutants are detected in nearly 40% of children. In advanced SM, including aggressive SM, SM with an associated hematological neoplasm, and mast cell leukemia, additional somatic mutations in other genes, such as SRSF2, JAK2, RUNX1, ASXL1, or RAS, may be detected. These drivers are more frequently detected in SM with an associated hematological neoplasm, particularly in male patients. Recently, hereditary alpha-tryptasemia has been identified as a genetic trait more prevalent in SM compared with healthy controls. Moreover, hereditary alpha-tryptasemia is more frequent in patients with SM with Hymenoptera venom allergy and severe mediator-related symptoms than in patients with SM without symptoms. On the basis of this knowledge, we propose a diagnostic algorithm in which genetic markers are applied together with clinical and histopathologic criteria to establish the diagnosis and prognosis in SM

Standards of pathology in the diagnosis of systemic mastocytosis: recommendations of the EU-US cooperative group

Pathology plays a central role in the diagnosis of systemic mastocytosis (SM), its delineation from other neoplasms and reactive conditions, and in monitoring of SM under therapy. The morphologic hallmark of SM is the accumulation of spindle-shaped, hypogranulated mast cells (MCs) in bone marrow (BM) and other extracutaneous tissues. Four of the 5 World Health Organization–defined diagnostic criteria (ie, compact MC aggregates [=major criterion]; atypical MC morphology; activating KIT point mutations; aberrant expression of CD25 and/or CD2 and/or CD30 in MCs [=minor criteria]) can be addressed by the pathologist. The final classification of SM variants as either BM mastocytosis, indolent SM, smoldering SM, aggressive SM (ASM), SM with an associated hematologic neoplasm (SM-AHN), or MC leukemia (MCL) has important prognostic significance and requires the integration of certain morphological, clinical, radiological, and biochemical data, referred to as B- and C-findings. Substantial diagnostic challenges may be posed to the pathologist and clinician especially in the so-called advanced SM variants, that is, ASM, MCL, and SM-AHN. In this article, updated recommendations of the EU-US Cooperative Group regarding standards of pathology in the diagnosis of SM, presented during the year 2020 Working Conference held in September in Vienna, are reported.T. I. George was supported by the ARUP Institute for Clinical and Experimental Pathology. K. Hartmann was supported by the Swiss National Science Foundation, grant number 310030_207705. D. D. Metcalfe, J. J. Lyons, and M. Carter were supported by the Division of Intramural Research, National Institutes of Allergic and Infectious Diseases, National Institutes of Health (NIH). The content is solely the responsibility of the authors and does not represent the official views of the NIH. P. Valent was supported by the Austrian Science Funds (FWF), projects F4701-B20 and F4704-B20

Standards of genetic testing in the diagnosis and prognostication of systemic mastocytosis in 2022: Recommendations of the EU-US cooperative group

Mastocytosis comprises rare heterogeneous diseases characterized by an increased accumulation of abnormal mast cells in various organs/tissues. The pathogenesis of mastocytosis is strongly linked to the presence of KIT-activating mutations. In systemic mastocytosis (SM), the most frequent mutation encountered is KIT p.D816V, whose presence constitutes one of the minor diagnostic criteria. Different techniques are used to search and quantify the KIT p.D816V mutant; however, allele-specific quantitative PCR and droplet digital PCR are today the most sensitive. The analysis of the KIT p.D816V allele burden has undeniable interest for diagnostic, prognostic, and therapeutic monitoring. The analysis of non–mast cell hematological compartments in SM is similarly important because KIT p.D816V multilineage involvement is associated with a worse prognosis. In addition, in advanced forms of SM, mutations in genes other than KIT are frequently identified and affect negatively disease outcome and response to therapy. Thus, combined quantitative and sensitive analysis of KIT mutations and next-generation sequencing of other recurrently involved myeloid genes make it possible to better characterize the extent of the affected cellular compartments and additional molecular aberrations, providing a more detailed overview of the complex mutational landscape of SM, in relation with the clinical heterogeneity of the disease. In this article, we report the latest recommendations of the EU-US Cooperative Group presented in September 2020 in Vienna during an international working conference, on the techniques we consider standard to detect and quantify the KIT p.D816V mutant in SM and additional myeloid mutations found in SM subtypes.D.D.M., J.J.L., and M.C.C. were supported by the Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health. P.V. was supported by the Austrian Science Fund (FWF) (grant nos. F4704-B20 and P32470-B)

Global Classification of Mast Cell Activation Disorders:An ICD-10-CM–Adjusted Proposal of the ECNM-AIM Consortium

Mast cell activation (MCA) is common and occurs in a number of pathologic conditions, including IgE-dependent and independent allergic reactions, atopic disorders, autoimmune processes, and mastocytosis. In a subset of patients, no underlying disease and no known trigger of MCA are found. When the symptoms are severe, systemic, and recurrent, and accompanied by a diagnostic increase in the serum tryptase level or other mast cell mediators, an MCA syndrome (MCAS) may be diagnosed. In these patients, the symptoms typically respond to drugs suppressing MCA, mediator production in mast cells, or mediator effects. In each case, diagnostic consensus criteria must be fulfilled to diagnose MCAS. In other patients, MCA may be local, less severe, or less acute, or may be suspected but not confirmed, so that the diagnostic criteria of MCAS are not fulfilled. In these patients, it may be difficult to prove MCA, for example, by measuring multiple mast cell mediators or basophil activation, the latter as a surrogate of IgE-dependent hypersensitivity. However, validated diagnostic criteria for implicating suspected MCA behind such conditions are lacking, even if some of these conditions have recently been assigned to an International Classification of Diseases-10-Clinical Modification code (ICD-10-CM). In this article, we discuss diagnostic features and criteria and propose a ICD-10-CM–adjusted classification for disorders associated with MCA, herein referred to as MCA disorders (MCADs), with special emphasis on the delineation between confirmed MCAS, MCAD not fulfilling MCAS criteria, and suspected MCAD that is not present. In addition, we discuss the discrimination between overt MCAD and predisposing conditions, such as atopic states, mastocytosis, and hereditary alpha tryptasemia.</p

Mutation analysis of "Endoglin" and "Activin receptor-like kinase" genes in German patients with hereditary hemorrhagic telangiectasia and the value of rapid genotyping using an allele-specific PCR-technique

<p>Abstract</p> <p>Background</p> <p>Hereditary hemorrhagic telangiectasia (HHT), also known as Rendu-Osler-Weber syndrome, is an autosomal dominant disorder which is clinically characterised by recurrent epistaxis, mucocutaneous telangiectasia and visceral arteriovenous malformations. Genetic linkage studies identified two genes primarily related to HHT: endoglin (<it>ENG</it>) on chromosome 9q33-34 and activin receptor-like kinase1 (<it>ACVRL1</it>) on chromosome 12q13. We have screened a total of 41 unselected German patients with the suspected diagnosis of HHT. Mutation analysis for the <it>ENG </it>and <it>ACVRL1 </it>genes in all patients was performed by PCR amplification. Sequences were then compared to the HHT database <url>http://www.hhtmutation.org</url> sequences of the <it>ENG </it>mRNA (accession no. BC014271.2) and the <it>ACVRL1 </it>mRNA (accession no. NM000020.1).</p> <p>Results</p> <p>We identified 15 different mutations in 18 cases by direct sequencing. Among these mutations, one novel <it>ENG </it>mutation could be detected which has not yet been described in the literature before. The genotype-phenotype correlation was consistent with a higher frequency of pulmonary arteriovenous malformations in patients with <it>ENG </it>mutations than in patients with <it>ACVRL1 </it>mutations in our collective.</p> <p>Conclusion</p> <p>For rapid genotyping of mutations and SNPs (single nucleotide polymorphisms) in <it>ENG </it>and <it>ACVRL1</it>, allele-specific PCR methods with sequence-specific primers (PCR-SSP) were established and their value analysed.</p

Current advances in the basic research and clinical management of sinonasal inverted papilloma (review)

Inverted papilloma (IP) is a benign sinonasal lesion that has a known propensity for recurrence, local aggressiveness and an association with transformation to squamous cell carcinoma. Due to the high rate of recurrence, association with malignancy and a tendency of multicentricity, the surgical approaches to treatment are controversial. Over the years there has been a slow evolution from aggressive (en bloc) resection by lateral rhinotomy to endoscopic techniques. This progress corresponds to the advances that have been made in endoscopic sinus surgery over the past 15 years. Technological advances have allowed the detection of sinonasal IP before its extension beyond the sinonasal region, thus enabling minimally invasive techniques to be used in the treatment of selected cases of IP. Differences in recurrence rates were not observed for endoscopic management as compared with lateral rhinotomy or sublabial degloving approaches. In terms of aetiology there is certain evidence that the presence of HPV in IP could be predictive of malignant transformation. Although IPs are monoclonal proliferations, they do not fit the profile of a prototypic precursor lesion. In contrast, an increased EGFR and TGF-alpha expression is associated with early events in IP carcinogenesis. Parameters such as hyperkeratosis, squamous epithelial hyperplasia and a high mitotic index are negative prognostic indicators, which could be useful in the future follow-up of patients with IP. Present literature should encourage us to recommend the use of a uniformly accepted staging system. The propensity for delayed recurrences and the maximal 13% incidence of malignant transformation mandates careful, long-term follow-up

Chondrosarcoma of the larynx and review of the literature

Chondrosarcoma (CS) of the larynx was first described in 1935. Cartilaginous tumours of the larynx are largely rare and there is little literature concerning them. Laryngeal CS manifest with a different pathological behaviour to other malignancies of the larynx and as such the treatment of these neoplasias are different. The purpose of this review is to present a detailed report of the laryngeal CS in recent literature. We present a case of laryngeal CS of the cricoid cartilage and a case of a sarcomatous neoplasm of the vocal cord as a potential differential diagnosis. Although representing a rare malignancy, the last decade has brought new insights in surgical treatment of laryngeal CS and subsequent reduction in recurrence rates, whereas progress in tumour biology and etiological agents is still scarce. We concentrate on new insights in classification, radiologic and pathologic features, and treatment modalities in the last two decades. Based on the literary evidence the authors recommend a conservative laryngeal function -preserving surgery. Total laryngectomy should be reserved to recurrent CS and rare cases of voluminous high-grade CS of the larynx

Regulation of matrix metalloproteinases (MMP)-2/-9 expression in eosinophilic chronic rhinosinusitis - Cell culture by interleukin-5 and-13?

Background: Eosinophils are a prominent immunological feature of chronic rhinosinusitis (CRS). Cytokines in the respiratory mucosa may be the key to upper airway pathophysiology. Matrix metalloproteinases (MMP) represent an entire group of Zn2+ dependent endopeptidases with the potential to alter the extracellular matrix (ECM). In this study epithelial cultures of CRS were treated with interleukin (IL)-5 or IL-13 and subsequent levels of metalloproteinases were determined. Materials and Methods: The cells for CRS culture were obtained from patients undergoing functional endoscopic sinus surgery. After 8-72 hours incubation with 0.2-0.4 ng/ml IL-5 or 3-6 ng/ml IL-13, the expression of the MMP-2 and -9 in the CRS cultures was analysed. Results: After 72 hours incubation with IL-5, the relative levels of MMP-2 showed no significant alteration in protein expression in comparison with the control groups. Incubation with IL-13 revealed a statistically insignificant decrease of the relative MMP-9 expression in ECRS compared to the control group (p>0.1). Conclusion: Alterations of MMP-2 and -9 expression may play a role in ECRS, but the association with IL-5 and IL-13 remains unclear