Tumor progression in melanocytic lesions : biological and diagnostic implications

Cutaneous malignant melanoma has an annual incidence of 5% in Sweden, with about 1500 cases diagnosed yearly. In routine histopathology there are diagnostic difficulties in the analysis of pigmented lesions. In spite of useful generalizations and criteria for the diagnosis of pigmented lesions, there are clinical as well as histological difficulties. To distinguish melanoma from Spitz nevi proliferation was assessed using antibodies against Ki-67 (MIB-1) and the anti-apoptotic protein BcI-2. My results clearly show that benign and dysplastic nevi in contrast to malignant lesions have a low proliferation. The majority (96%) of the mm expressed Ki-67 as well as Bcl-2, while this was only seen in 6% of the Spitz nevi. IGF-1R is required for the establishment and maintenance of the transformed phenotype and is also important as an anti-apoptotic regulator in melanoma cells. By Western blotting and binding analysis I could confirm IGF-1R expression in 4 several melanoma cell lines. Two ways of blocking the IGF-1R function were used. For the translocation of the receptor to the cell surface N-linked glycosylation is essential. Cell growth arrest and cell death were induced by the N-linked glycosylation blocker Tunicamycin. A similar effect was obtained by the [alpha]IR-3 antibody, which blocks the binding domain of IGF-1R. These treatments resulted in apoptosis. Primary melanoma tissues and tissues from melanoma metastases, obtained from lymph nodes of patients with advanced mm, were also found to express the IGF-1R. However, none of benign nevi used for comparison stained positive. I also noted that there was an inverse correlation between the expression of IGF-1R. and the frequency of apoptotic cells, which suggest that IGF-1R. has also an anti-apoptotic effect in vivo. Cdk inhibitors may be important in controlling cellular proliferation. To evaluate other possible mechanisms in IGF-1R. dependent control of proliferation, the influence of p27 Kip1 and its related cyclins were of interest as my analyses have revealed an inverse relation between p27 Kip1 and expression of IGF-1R. in melanoma cells. Both Tunicamycin and [alpha]IR-3 caused substantial redistribution of p27 from cyclin DI to cyclin E and cyclin A. These effects may underlie the growth-inhibitory and apoptotic; effect of inhibition of IGF-1R. in melanoma cells. Previous clinical studies have indicated that the estrogen receptor blocker, Tamoxifen (TAM) has some favorable effects on the clinical outcome of mm. There is support from studies on breast cancer that TAM interferes with the IGF- 1 pathway by increasing the release of insulin-like growth factor binding proteins (IGFBPs). I showed that TAM induced cytotoxicity in several nun cell lines, which did not express ER. I could, however, confirm that TAM did not affect the IGFBPs. Instead, it seems that TAM inhibits autophosphorylation of the [beta]-subunit of the IGF- 1 receptor. This may indicate the role of TAM as an inhibitor of tyrosine kinases and a potential therapeutical approach for advanced mm

The prognostic implications of Notch1, Hes1, Ascl1, and DLL3 protein expression in SCLC patients receiving platinum-based chemotherapy.

ObjectivesThe aim was to analyse the tumor expression of Notch1, Hes1, Ascl1, and DLL3 in Small-Cell Lung Cancer (SCLC) and each such biomarker's potential association with clinical characteristics and prognosis after platinum-doublet chemotherapy (PDCT).Material and methodsThe protein expression of the biomarkers was evaluated using immunohistochemistry. Patients were categorized according to their sensitivity to first line PDCT: with a Progression-free survival (PFS) ≥ 3 months after completion of treatment considered "sensitive" and Results and conclusionThe study included 46 patients, with 21 and 25 of the patients having "sensitive" and "refractory" disease, respectively. The majority of patients had a high DLL3 expression (n = 38), while a minority had Notch 1-high expression (n = 10). The chi-square test showed that there was a statistically significant negative association between Notch1 and Ascl1 expression (p = 0.013). The overall survival for patients with Notch1- high vs. low expression was 8.1 vs. 12.4 months, respectively (p = 0.036). Notch1 expression was an independent prognostic factor in the multivariate analysis (p = 0.02). No other biomarker showed any prognostic impact in this highly selected SCLC cohort. DLL3 is highly expressed in the majority of advanced staged SCLC cases, as expected. In the same patient population, Notch1 expression might have a potential prognostic implication, by driving a non-neuroendocrine differentiation process. Given the small number of cases with Notch1 high expression, the results of this study needs to be confirmed on a larger cohort

Caspase-2 is a mediator of apoptotic signaling in response to gemtuzumab ozogamicin in acute myeloid leukemia

The antibody conjugate gemtuzumab ozogamicin (GO; Mylotarg((R))) provides targeted therapy of acute myeloid leukemia (AML), with recent approvals for patients with CD33-positive disease at diagnosis or relapse, as monotherapy or combined with chemotherapeutics. While its clinical efficacy is well documented, the molecular routes by which GO induces AML cell death warrant further analyses. We have earlier reported that this process is initiated via mitochondria-mediated caspase activation. Here we provide additional data, focusing on the involvement of caspase-2 in this mechanism. We show that this enzyme plays an important role in triggering apoptotic death of human AML cells after exposure to GO or its active moiety calicheamicin. Accordingly, the caspase-2 inhibitor z-VDVAD-fmk reduced GO-induced caspase-3 activation. This finding was validated with shRNA and siRNA targeting caspase-2, resulting in reduced caspase-3 activation and cleavage of poly [ADP-ribose] polymerase 1 (PARP-1). We previously demonstrated that GO-induced apoptosis included a conformational change of Bax into a pro-apoptotic state. Present data reveal that GO-treatment also induced Bid cleavage, which was partially reduced by caspase-2 specific inhibition while the effect on GO-induced Bax conformational change remained unaltered. In mononuclear cells isolated from AML patients that responded to GO treatment in vitro, processing of caspase-2 was evident, whereas in cells from an AML patient refractory to treatment no such processing was seen. When assessing diagnostic samples from 22 AML patients, who all entered complete remission (CR) following anthracycline-based induction therapy, and comparing patients with long versus those with short CR duration no significant differences in baseline caspase-2 or caspase-3 full-length protein expression levels were found. In summary, we demonstrate that GO triggers caspase-2 cleavage in human AML cells and that the subsequent apoptosis of these cells in part relies on caspase-2. These findings may have future clinical implications

Interobserver Variability of Histopathological Prognostic Parameters in Cutaneous Malignant Melanoma : Impact on Patient Management

Clinical management of primary cutaneous melanomas is based on histopathological staging of the tumour. The aim of this study was to investigate, in a non-selected population in clinical practice, the agreement rate between general pathologists and pathologists experienced in melanoma in terms of the evaluation of histopathological prognostic parameters in cutaneous malignant melanomas, and to what extent the putative variability affected clinical management. A total of 234 cases of invasive cutaneous malignant melanoma were included in the study from the Stockholm-Gotland Healthcare Region in Sweden. Overall interobserver variability between a general pathologist and an expert review was 68.8-84.8%. Approximately 15.5% of melanomas <= 1 mm were re-classified either as melanoma in situ or melanomas >1 mm after review. In conclusion, review by a pathologist experienced in melanoma resulted in a change in recommendations about surgical excision margins and/or sentinel node biopsy in subgroups of Ti melanomas

Playing as an early-childhood promotion of communication skills

Darba autore: Agita Kaža Darba tēma: Rotaļa kā saskarsmes prasmju sekmētāja agrās bērnības vecumā Darba vadītājs: Prof. Dr. psych.Tija Zīriņa Pētījuma mērķis: Pētīt 2 – 3 gadīgu bērnu saskarsmes prasmes rotaļās Pētījuma saturs: darba teorētiskās daļas pirmajā nodaļā tiek aplūkotas un analizētas dažādu pedagogu, psihologu atziņas par saskarsmi, kā veidojas saskarsme pirmsskolā. Darba otrajā nodaļā tiek pētīta agrās bērnības saskarsmju prasmju sekmēšanā rotaļās. Nodaļas saturs lielākoties balstās uz praktisko pētījumu. Pētījumā piedalās agrās bērnības vecuma 16 bērni. Pētījumā tiek organizētas rotaļas ar kuru palīdzību bērni veicina sekmīgu saskarsmes prasmju veidošanos. Secinājumos apkopota nozīmīgākā informācija saskarsmes prasmju attīstību rotaļā. Atslēgas vārdi: saskarsme, saskarsmes prasmju attīstība, rotaļa, pirmsskola, bērni. Darba apjoms: darbs sastāv no 4 nodaļām un 5 apakšnodaļām, tam ir 55 lapas. Darbā aplūkojamas 2 tabulas, 2 attēli un pielikums, kurā ir redzams 21 attēls, izmantotās literatūras saraksts sastāv no 45 avotiem.Author of the work: Agita Kaza Theme of the work: Playing as an early-childhood promotion of communication skills Supervisor of the work: Prof. Dr. psych. Tija Zirina The aim of the research: To study 2-3-year-olds' skills in games Study content: the first chapter of the theoretical part of the work looks at and analyzes the knowledge of various educators, psychologists, about how they come into contact in preschool. The second section of work examines the promotion of early childhood communication skills in games. The content of the chapter is largely based on the practical study. 16 children of the age of early childhood are involved in the study. The study organizes games to help children develop successful communication skills. The conclusions summarise key information on the development of communication skills in play. Keywords: communication, development of communication skills, playing, preschool, children. Scope of the work: the work consists of 4 chapters and 5 subdivisions, with 55 pages. The work includes 2 tables, 2 pictures and an attachment showing 21 figures, a list of authorities used consisting of 45 sources

Protein profiling of fine-needle aspirates reveals subtype-associated immune signatures and involvement of chemokines in breast cancer

There are increasing demands for informative cancer biomarkers, accessible via minimally invasive procedures, both for initial diagnostics and for follow-up of personalized cancer therapy, including immunotherapy. Fine-needle aspiration (FNA) biopsy provides ready access to relevant tissue samples; however, the minute amounts of sample require sensitive multiplex molecular analysis to be of clinical biomarker utility. We have applied proximity extension assays (PEA) to analyze 167 proteins in FNA samples from patients with breast cancer (BC; n = 25) and benign lesions (n = 32). We demonstrate that the FNA BC samples could be divided into two main clusters, characterized by differences in expression levels of the estrogen receptor (ER) and the proliferation marker Ki67. This clustering corresponded to some extent to established BC subtypes. Our analysis also revealed several proteins whose expression levels differed between BC and benign lesions (e.g., CA9, GZMB, IL-6, VEGFA, CXCL11, PDL1, and PCD1), as well as several chemokines correlating with ER and Ki67 status (e.g., CCL4, CCL8, CCL20, CXCL8, CXCL9, and CXCL17). Finally, we also identified three signatures that could predict Ki67 status, ER status, and tumor grade, respectively, based on a small subset of proteins, which was dominated by chemokines. To our knowledge, expression profiles of CCL13 in benign lesions and BC have not previously been described but were shown herein to correlate with proliferation (P = 0.00095), suggesting a role in advanced BC. Given the broad functional range of the proteins analyzed, immune-related proteins were overrepresented among the observed alterations. Our pilot study supports the emerging role of chemokines in BC progression. Due to the minimally traumatic sampling and clinically important molecular information for therapeutic decisions, this methodology is promising for future immunoscoring and monitoring of treatment efficacy in BC

Multiplex immune protein profiling of fine‐needle aspirates from patients with non‐small‐cell lung cancer reveals signatures associated with PD‐L1 expression and tumor stage

Biomarker signatures identified through minimally invasive procedures already at diagnosis of non‐small‐cell lung cancer (NSCLC) could help to guide treatment with immune checkpoint inhibitors (ICI). Here, we performed multiplex profiling of immune‐related proteins in fine‐needle aspirate (FNA) samples of thoracic lesions from patients with NSCLC to assess PD‐L1 expression and identify related protein signatures. Transthoracic FNA samples from 14 patients were subjected to multiplex antibody‐based profiling by proximity extension assay (PEA). PEA profiling employed protein panels relevant to immune and tumor signaling and was followed by Qlucore® Omics Explorer analysis. All lesions analyzed were NSCLC adenocarcinomas, and PEA profiles could be used to monitor 163 proteins in all but one sample. Multiple key immune signaling components (including CD73, granzyme A, and chemokines CCL3 and CCL23) were identified and expression of several of these proteins (e.g., CCL3 and CCL23) correlated to PD‐L1 expression. We also found EphA2, a marker previously linked to inferior NSCLC prognosis, to correlate to PD‐L1 expression. Our identified protein signatures related to stage included, among others, CXCL10 and IL12RB1. We conclude that transthoracic FNA allows for extensive immune and tumor protein profiling with assessment of putative biomarkers of important for ICI treatment selection in NSCLC

Quantitative Proteomics Profiling of Primary Lung Adenocarcinoma Tumors Reveals Functional Perturbations in Tumor Metabolism

In this study, we have analyzed human primary lung adenocarcinoma tumors using global mass spectrometry to elucidate the biological mechanisms behind relapse post surgery. In total, we identified over 3000 proteins with high confidence. Supervised multivariate analysis was used to select 132 proteins separating the prognostic groups. Based on in-depth bioinformatics analysis, we hypothesized that the tumors with poor prognosis had a higher glycolytic activity and HIF activation. By measuring the bioenergetic cellular index of the tumors, we could detect a higher dependency of glycolysis among the tumors with poor prognosis. Further, we could also detect an up-regulation of HIF1 alpha mRNA expression in tumors with early relapse. Finally, we selected three proteins that were upregulated in the poor prognosis group (cathepsin D, ENO1, and VDAC1) to confirm that the proteins indeed originated from the tumor and not from a stromal or inflammatory component. Overall, these findings show how in-depth analysis of clinical material can lead to an increased understanding of the molecular mechanisms behind tumor progression

Quantitative Proteomics Profiling of Primary Lung Adenocarcinoma Tumors Reveals Functional Perturbations in Tumor Metabolism

In this study, we have analyzed human primary lung adenocarcinoma tumors using global mass spectrometry to elucidate the biological mechanisms behind relapse post surgery. In total, we identified over 3000 proteins with high confidence. Supervised multivariate analysis was used to select 132 proteins separating the prognostic groups. Based on in-depth bioinformatics analysis, we hypothesized that the tumors with poor prognosis had a higher glycolytic activity and HIF activation. By measuring the bioenergetic cellular index of the tumors, we could detect a higher dependency of glycolysis among the tumors with poor prognosis. Further, we could also detect an up-regulation of HIF1α mRNA expression in tumors with early relapse. Finally, we selected three proteins that were upregulated in the poor prognosis group (cathepsin D, ENO1, and VDAC1) to confirm that the proteins indeed originated from the tumor and not from a stromal or inflammatory component. Overall, these findings show how in-depth analysis of clinical material can lead to an increased understanding of the molecular mechanisms behind tumor progression