Pathology & Oncology Research最新文献

Photobleaching of immunofluorescence signal is a well-known phenomenon, however, its impact on derived parameters characterizing number and shape of different cell types in tissue sections is less understood. Our aim was to determine whether the duration of illumination and the type of fluorophore (Alexa Fluor 546 (A546), and Alexa Fluor 488 Plus (A488)) can influence the acquired morphometric parameters of cells in the nervous system. Immunofluorescent staining of microglia and neurons was performed on mouse spinal cord sections. Mean color intensity in a field of view, number of detectable neuronal cell profiles, partial coverage of microglial profiles, and fractal geometrical parameters were determined. All measurements were made using epifluorescence microscopy with identical acquisition parameters. Most of the measured parameters suffered significant alternation after 30-60 s of illumination. The data-altering effect of photobleaching was most prominent in the case of mean fluorescent intensity. Thus, while immunofluorescent staining is useful for co-localizing different groups of cells, cell-specific quantitative morphological measurements require photostable staining. Possibility of the combination of these methods on the same section in order to achieve multi-channel localization without photobleaching is exemplified.

Pazopanib is a tyrosine-kinase inhibitor also used for the treatment of advanced soft tissue sarcomas. Our retrospective study analyzed real-world data of stage 4 sarcoma patients treated with pazopanib in our department in the past 10 years. Data were collected from the Medworks medical system, which is used for daily work in our center. A total of 99 patients were included: 46 men and 53 women The median age at the diagnosis was 49.8 years. The most common histological subtypes were leiomyosarcoma and synovial sarcoma. All patients received 800 mg of pazopanib per day, which was reduced to 400 mg in the event of toxicity. Treatment was continued until disease progression or unmanageable toxicity. The primary endpoint of the study was progression-free survival and the secondary endpoints were overall survival, overall response rate and disease control rate. The results in relation to demographic data, previous treatments, localizations of primary tumors and metastasis and histological subtypes were analyzed. In our center pazopanib was most frequently used in the third line. In total, 61 patients received perioperative therapy; the most common regimen used in the metastatic setting was VIP. Median PFS and OS were 3 months and 7 months, respectively. ORR was 14% and DCR was 40.45%. Dose reductions were necessary during the treatment of 56 patients. Hematological toxicity was detected in 23% of cases, with the most frequent events being grade 1 thrombocytopenia and grade 2 leukocytopenia. Non-hematological adverse events were documented in half of the patients. Pazopanib was more effective in earlier lines of treatment. Compared to the PALETTE phase 3 trial more patients received perioperative therapy, median PFS and OS were shorter (3 months vs. 4.6 months and 7 months vs. 11.9 months) and ORR was higher (14% vs. 9%) in our patient population. Dose reductions were more frequent in our center. Pazopanib is a therapeutic option for the treatment of advanced soft tissue sarcoma, also according to real-world data. Further investigations are needed to select patients who can benefit the most from pazopanib and to determine the most appropriate sequence of therapy.

The randomized controlled pivotal phase 3 study evaluated efficacy and safety of neoadjuvant complex biologic, Leukocyte Interleukin Injection (LI), administered for 3 consecutive weeks pre-surgery, in treatment naïve resectable locally advanced primary squamous cell carcinoma of oral cavity and soft palate. Randomization 3:1:3 to LI+/-CIZ (cyclophosphamide, indomethacin, and zinc)+SOC, or SOC (standard of care) alone. LI-treated patients received 400 IU (as interleukin-2 equivalent; 200 IU peritumorally, 200 IU perilymphatically) sequentially, daily 5 days/week for 3 weeks before surgery. All subjects were to receive SOC. Post-surgery, patients with low risk for recurrence were to receive radiotherapy, while those with high risk received concurrent chemoradiotherapy. Median follow-up was 56 months. There were 923 ITT (Intent-to-Treat) subjects (380 ITT low-risk and 467 ITT high-risk). Pre-surgery objective early response (45 objective early responders; 5 complete responses [CRs], 40 partial responses [PRs], confirmed by pathology at surgery. LI (+/- CIZ) had 8.5% objective early responders (45/529 ITT) and 16% objective early responders (34/212 ITT low-risk) vs. no reported SOC objective early responders (0/394 ITT). Objective early responders significantly lowered death rate to 22.2% (ITT LI-treated), 12.5% (ITT low-risk LI + CIZ + SOC), while the ITT low-risk SOC death rate was 48.7%. Thus, objective early response impacted overall survival (OS); proportional hazard ratios were 0.348 (95% CI: 0.152-0.801) for ITT low-risk LI-treated, 0.246 (95% CI: 0.077-0.787) for ITT low-risk LI + CIZ + SOC. ITT low-risk LI + CIZ + SOC demonstrated significant OS advantage vs. ITT low-risk SOC (unstratified log-rank p = 0.048; Cox hazard ratio = 0.68; 95% CI: 0.48-0.95, Wald p = 0.024 [controlling for tumor stage, tumor location, and geographic region]). Absolute OS advantage increased over time for ITT low-risk (LI + CIZ + SOC)-treated vs. ITT low-risk SOC: reaching 14.1% (62.7% vs. 48.6%) at 60 months, with 46.5 months median OS advantage (101.7 months vs. 55.2 months), respectively. Quality of life benefit for complete responders sustained for >3 years post LI treatment. Percent treatment-emergent adverse events were comparable among all treated groups. No excess safety issues were reported for LI over SOC alone post-surgery. NCT01265849, EUDRA:2010-019952-35.

Introduction: Immunotherapy has made a significant improvement in the treatment of patients with non-small cell lung cancer (NSCLC). It has a role in boosting the immune system, so it can fight cancer cells. Sometimes, this mechanism can lead to an overstimulation or misdirection of immune response, so it can act against the body itself. One of the organs most affected by this reaction is the thyroid gland, and there is no definitive explanation of the causes of this adverse event.

Material and methods: In this retrospective observational study, we enrolled 103 patients with NSCLC and high PD-L1 expression (>= 50%) who were treated in our Clinic for pulmonology, University Clinical Center of Serbia, using Pembrolizumab as the first-line therapy.

Results: Data analysis showed that 41 (39.81%) of 103 patients in our study had an adverse event of immunotherapy, and 21 of them had autoimmune thyroiditis (20.39%). Of all the patients, 19 of them were treated for chronic obstructive pulmonary disease (COPD) before the onset of Pembrolizumab. During treatment, eight of these patients developed thyroid dysfunction. Patients with COPD were at increased risk of developing autoimmune thyroiditis compared to non-COPD patients (OR 3.9 95% CI 1.135-13.260, p = 0.0227).

Conclusion: Our study showed that patients dealing with COPD have a 3.9 times greater risk of developing autoimmune thyroiditis as an adverse event during Pembrolizumab treatment compared with patients without COPD.

Comprehensive genomic profiling (CGP) is becoming an increasingly important tool in the clinical management of different tumours, but there is still very limited data available on its usefulness from a therapeutic point of view in mesenchymal tumours. Between January 2022 and September 2024, we performed CGP analysis with means of Oncomine Comprehensive Assay Plus (OCAplus) on 94 malignant mesenchymal tumours. The analysis covered more than 500 unique genes for single-gene and multigene biomarker insights, including tumour mutational burden (TMB) and homologous recombination deficiency (HRD). Genomic DNA and total RNA were extracted from formalin-fixed paraffin-embedded tissue blocks. Twenty-four out of 94 patients (25.5%) had potentially actionable alterations: 17 (18%) had specific genetic alterations suitable for targeted therapies, 4 (4.2%) had a high TMB (>10 mut/Mb), and 5 (5.3%) had a high HRD score >15). One additional patient had BRCA1 mutation, but the HRD score was low. Three patients received targeted therapy: one patient with a CDK4-amplified tumour (dedifferentiated liposarcoma) received CDK4 inhibitor therapy, two patients with angiosarcoma showing high TMB received immune checkpoint inhibitor therapy, and one patient with a uterine leiomyosarcoma and high HRD score received PARP inhibitor therapy. In addition, two patients with malignant phyllodes tumours received multi-thyrosine kinase inhibitor therapy. In three cases, there was refinement or reassignment of the diagnosis, based on the CGP findings. Our results demonstrate that CGP can provide useful additional information and can be beneficial in the clinical management of patients with mesenchymal tumours.

Sarcomas with an EWSR1::POU2AF3(COLCA2) fusion are a very recently described entity of preferentially sinonasal origin and with undifferentiated round/spindle cell morphology. We established a novel cell line (PF1095) carrying a EWSR1::POU2AF3 fusion from the malignant pleural effusion of a 25-year-old sarcoma patient. The patient was first diagnosed with poorly differentiated neuroendocrine carcinoma based on tumor cell morphology and positivity to markers such as EMA, synaptophysin, and CD56. Later, the EWSR1 translocation was identified in the tumor cells with unknown partners and the patient received chemotherapy according to the Ewing 2008 protocol in combination with surgery and proton beam radiotherapy. At the time of cell line establishment, the disease progressed to pleural sarcomatosis with pleural effusion. In the cell line, we identified POU2AF3 as a fusion partner of EWSR1 and a TP53 frameshift deletion. Next, we determined the sensitivity of PF1095 cells to the currently approved chemotherapies in comparison to two conventional Ewing sarcoma lines (EW-7 and MHH-ES1) with the two most frequent EWSR::FLI1 fusions. Finally, we tested potential new combination therapies. We performed cell viability, proliferation, and cell cycle assays. We found that the proliferation rate of PF1095 cells was much slower than the EWSR1::FLI1 fusion lines and they also had a lower sensitivity to both irinotecan and doxorubicin treatment. Expression level of SLFN11, a predictor of sensitivity to DNA damaging agents, was also lower in PF1095 cells. Combination treatment with the PARP inhibitors olaparib and irinotecan or doxorubicin synergistically reduced cell viability and induced cell death and cell cycle arrest. This unique cell model provides an opportunity to test therapeutic approaches preclinically for this novel and aggressive sarcoma entity.

Introduction: Polycythemia indicates the pathological increase in the number of red blood cells and the rise of hematocrit values. Polyglobulia can be of primary or secondary origin, with the most common primary polycythemia being a myeloproliferative neoplasm, polycythemia vera. Polyglobulia patients may develop cardiovascular complications and thromboembolic events. The gold standard of first-line treatment in polycythemia vera is phlebotomy, which is indicated to keep the hematocrit value below 0.45. Until now the goal to be achieved in secondary polyglobulia has been similar. In secondary polyglobulia this rule of thumb needs to be re-evaluated as shown by the example of two patients suffering from different rare, genetically determined polyglobulias. In our article we present the case of these two patients and discuss the diagnostic and therapeutic principles to be applied in patients with rare, genetically determined polyglobulias.

Patients and methods: After completing the usual diagnostic algorithm for polyglobulia no cause could be identified in two of our male patients. Therefore, we set out to perform whole exome sequencing in both patients. Our analysis did not include copy number analysis.

Results: In Patient 1 the p.Ser179Pro variant in the VHL gene was detected in the homozygous state, which is classified as likely pathogenic according to the ACMG guidelines. Homozygous VHL mutations are implicated in Chuvash polycythemia. Segregation analysis was declined by the family. In Patient 2 the PKLR gene p.His306Gln variant was detected in the heterozygous form. The gene plays a role in pyruvate metabolism. Family screening did not detect this variant in healthy family members.

Discussion: We identified rare, possibly pathogenic genetic variants in two patients with polyglobulia and as a consequence of the genetic diagnosis we implemented individualized patient monitoring. We recommend the utilization of high-throughput genomic testing in cases with unexplained polyglobulia.

Background: Alectinib is effective in extending the survival of patients with anaplastic lymphoma kinase (ALK)-positive non-small cell lung cancer (NSCLC) and generally has manageable side effects. However, intestinal ulcers and colitis are rare but serious adverse reactions linked to Alectinib, meriting further investigation into their causes.

Case presentation: We report the case of a 62-year-old woman with NSCLC and brain metastases, who tested positive for ALK. She had been treated with Alectinib for nearly 4 years. The patient experienced diarrhea for 4 days, and a subsequent colonoscopy revealed pancolitis along with multiple ulcers in the terminal ileum and ileocecal valve. Given the severity of these symptoms, classified as a grade 3 adverse event by the Common Terminology Criteria for Adverse Events (CTCAE), Alectinib was discontinued. Treatment with oral enteric-coated Mesalazine tablets led to a resolution of the diarrhea and a significant improvement in the pancolitis and ulcers upon follow-up. The patient's anticancer therapy was subsequently switched to Ceritinib capsules. At follow-up, she demonstrated a stable tumor condition with no recurrence of intestinal ulcers or colitis.

Conclusion: To our knowledge, this is the first reported case of intestinal ulceration and colitis induced by Alectinib. Although such adverse events are exceedingly rare, they require vigilant monitoring in clinical practice. Decisions on continuing with Alectinib should consider the severity of side effects, classified by CTCAE grade. For managing these specific adverse events, oral Mesalazine enteric-coated tablets appear to be an effective treatment option.

Purpose: This study aims to evaluate whether survival outcomes for GIST patients have improved over the past decades and to identify the specific patient subgroups that have benefited from advances in treatment.

Patients and methods: A total of 4,127 GIST patients diagnosed between January 1980, and December 2019, were included in this study using data from the Surveillance, Epidemiology, and End Results (SEER)-9 Registries. Survival differences among GIST patients were analyzed across five time periods (1980-1999, 2000-2004, 2005-2009, 2010-2014, and 2015-2019) and within demographic, neoplastic, temporal, economic, and geographic categories using the log-rank test. Multivariable Cox regression models were employed to identify risk factors associated with GIST-specific survival. Associations between time periods and GIST-specific mortality (TSM) were examined using a multivariable Cox regression model.

Results: Survival outcomes for GIST patients significantly improved in the 2000-2009 period but showed no substantial improvement in the 2010-2019 period. After adjusting for age, gender, tumor location, ethnicity, tumor stage, median household income, and geographic area, the multivariable Cox regression models revealed that older age (≥65 years) (HR = 1.977, 95% CI = 1.470-2.657), tumors located outside the gastrointestinal tract (HR = 1.505, 95% CI = 1.267-1.786), regional lesions (HR = 2.225, 95% CI = 1.828-2.708), and distant lesions (HR = 5.177, 95% CI = 4.417-6.069) were independent risk factors for TSM (p < 0.05). After adjusting for time periods and age, gender, tumor location, tumor stage, median household income, patients in 2000-2004 (HR = 0.662, 95% CI = 0.523-0.839), 2005-2009 (HR = 0.431, 95% CI = 0.339-0.549), 2010-2014 (HR = 0.437, 95% CI = 0.341-0.561), and 2015-2019 (HR = 0.365, 95% CI = 0.273-0.489) had a significantly lower risk of TSM than patients in 1980-1999 (p < 0.05). Similarly, patients in 2005-2009 (HR = 0.661, 95% CI = 0.555-0.788), 2010-2014 (HR = 0.696, 95% CI = 0.578-0.838), and 2015-2019 (HR = 0.607, 95% CI = 0.476-0.773) also had a significantly lower risk of TSM than patients in 2000-2004 (p < 0.05). However, patients in 2010-2014 (HR = 1.042, 5% CI = 0.863-1.258) and 2015-2019 (HR = 0.945, 95% CI = 0.734-1.216) did not have a significantly lower risk of TSM compared to patients in 2005-2009 (p > 0.05).

Conclusion: GIST survival has significantly improved during the period 2000-2009 but showed no substantial improvement in 2010-2019, with the turning point for lower risk of TSM being 2005. Innovative strategies are needed to further improve survival outcomes for GIST patients, particularly for older patients and those with tumors originating outside the gastrointestinal tract.

Giant cell tumour of bone (GCTB) is viewed as a benign, locally aggressive primary bone tumour with metastatic potential. Current management is surgery with bone curettage or resection and systemic therapy with denosumab. Diagnosis is confirmed histologically prior to surgery, with staging for pulmonary disease, as pulmonary metastases (PM) reportedly occur in <8%. This study aimed to assess incidence, surveillance and management of PM in patients with GCTB, with histopathological review. A retrospective audit of the Oxford bone tumour registry was performed from January 2014 - October 2023. Inclusion criterion was histological confirmation of GCTB. Exclusion criteria were incomplete medical, imaging or histology records, or referral for secondary MDT opinion for diagnosis. From an initial group of 126 GCTB patients, 83 patients met the full selection criteria. Pulmonary metastases were identified in 11 patients. Three with PM were excluded on histopathological review as being giant cell rich osteosarcoma rather than metastatic GCTB. This left 8 (9.6%) patients, one had PM at presentation and seven at follow-up between 2 and 42 months. Two were histologically confirmed after cardiothoracic surgery and biopsy, six radiologically diagnosed. Three (37.5%) patients with PM have died (between 1 and 12 months after confirmed PM), five are alive with stable disease. Seven (87.5%) of patients with pulmonary disease were treated with denosumab/chemotherapy (three before, four after pulmonary diagnosis). Five (62.5%) with pulmonary disease had recurrence of local disease requiring further surgery. Local recurrence was an independent risk factor for PM on statistical analysis. GCTB may present with PM, but more commonly, metastasis occurs after surgery, presenting on surveillance and can progress. There were no distinct differences in histopathological appearance between patients with GCTB that developed PM and those that did not, therefore morphological features of the tumour cannot be currently used to predict tumour behaviour. PM can behave aggressively, necessitating identifying histological markers to recognise patients at risk of metastatic GCTB, for example, through mRNA single cell analysis. We propose GCTB patients with PM receive regular chest surveillance with PET scan and/or CT to monitor disease progression, and a multi-centre audit of GCTB outcome undertaken to further define optimal clinical management.