Subject(s)
Solitary Fibrous Tumor, Pleural , Humans , Pleural Neoplasms/surgery , Pleural Neoplasms/pathology , Pleural Neoplasms/diagnostic imaging , Pleural Neoplasms/diagnosis , Solitary Fibrous Tumor, Pleural/surgery , Solitary Fibrous Tumor, Pleural/pathology , Solitary Fibrous Tumor, Pleural/diagnostic imaging , Solitary Fibrous Tumor, Pleural/diagnosis , Tomography, X-Ray Computed , Treatment OutcomeABSTRACT
PURPOSE: Medullary thyroid carcinoma (MTC) in MEN2B syndrome is associated with germline RET mutation. Patients harboring de novo mutations are usually diagnosed at more advanced disease stages. We present a young woman with Met918Th mutation diagnosed with stage IV MTC at age 10 years. METHODS: The disease progressed despite total thyroidectomy and multiple surgical interventions for cervical lymph node recurrences, leading to distant metastases in the fifth year after the initial diagnosis. Subsequently, she underwent five different types of tyrosine kinase inhibitor (TKI) treatments. The 17-year disease course was divided into periods defined by four surgical interventions and sequential treatment intervals with four multikinase (sunitinib, vandetanib, cabozantinib, and lenvatinib) and one RET-selective TKI (selpercatinib). Tumor growth for different phases of spontaneous development and drug treatment intervals was characterized by changes in serial log-transformed calcitonin measurements (n = 114). RESULTS: Three operations (one for calcitonin-producing adrenal pheochromocytoma) were associated with drops in calcitonin levels. All of the nonselective TKIs were stopped due to adverse effects. As reflected by the negative calcitonin doubling rate, the best treatment response was observed with selpercatinib, which was associated with an initial large drop followed by a decreasing calcitonin trajectory over 514 days without any major side effects. CONCLUSION: This case of MEN2B medullary thyroid cancer with long-term survival presents how the effectiveness of different treatment modalities can be estimated using log-transformed calcitonin levels. Furthermore, our experience supports the view that serial calcitonin measurements may be more sensitive than radiological follow-up in advanced MTC. Our patient also represents a new case of rarely reported calcitonin-producing pheochromocytomas.
Subject(s)
Calcitonin , Carcinoma, Neuroendocrine , Multiple Endocrine Neoplasia Type 2b , Thyroid Neoplasms , Humans , Calcitonin/blood , Calcitonin/therapeutic use , Thyroid Neoplasms/blood , Thyroid Neoplasms/drug therapy , Thyroid Neoplasms/genetics , Thyroid Neoplasms/pathology , Female , Multiple Endocrine Neoplasia Type 2b/genetics , Multiple Endocrine Neoplasia Type 2b/blood , Carcinoma, Neuroendocrine/drug therapy , Carcinoma, Neuroendocrine/blood , Carcinoma, Neuroendocrine/genetics , Proto-Oncogene Proteins c-ret/genetics , Protein Kinase Inhibitors/therapeutic useABSTRACT
INTRODUCTION: Dedifferentiated liposarcoma (DDLPS) is a common form of liposarcoma with challenging treatment modalities. Pan-TRK immunopositivity can be often observed without NTRK gene fusion in soft tissue sarcomas with myogenic differentiation. Expression and the role of NTRK in DDLPS are under-studied. We sought to identify activating mutations of the NTRK genes. MATERIALS AND METHODS: 131 DDLPS patients were selected for pan-TRK immunohistochemistry and positive cases were analyzed by Sanger sequencing for NTRK1, NTRK2 and NTRK3 genes. Functional assays were performed using a lentiviral transduction system to study the effect of NTRK variants in fibroblast, immortalized fibroblast, and dedifferentiated liposarcoma cell lines. RESULTS: Out of the 131 DDLPS cases, 75 immunohistochemical staining positive cases, 46 were successfully Sanger sequenced. A recurrent somatic mutation pair in cis position (NGS) of the NTRK1 c.1810C>T (p.H604Y) and c.1838G>T (p.G613V) was identified in six cases (13%) that have never been reported in DDLPS. NTRK fusions were excluded in all six cases by FISH and NGS. The phospho-AKT immunopositivity among the six mutated cases suggested downstream activation of the NTRK signaling pathway. Functional assays showed no transforming effects, but resistance to first- and second-line TRK inhibitors of the p.G613V and p.H604Y variant. CONCLUSIONS: We detected (de novo/somatic) missense mutation variants in cis position of the NTRK1 gene in a subset of DDLPS indicating modifying mutations that may contribute to tumorigenesis in a subset of DDLPS. These variants beget resistance to TRK inhibitors indicating an interesting biomarker for other studies with TRK inhibitors.
Subject(s)
Liposarcoma , Sarcoma , Soft Tissue Neoplasms , Humans , Liposarcoma/genetics , Mutation , Oncogene Proteins, Fusion/genetics , Receptor, trkA/genetics , Sarcoma/genetics , Soft Tissue Neoplasms/geneticsABSTRACT
Alterations in mTOR signalling molecules, including RICTOR amplification, have been previously described in many cancers, particularly associated with poor prognosis. In this study, RICTOR copy number variation (CNV) results of diagnostic next-generation sequencing (NGS) were analysed in 420 various human malignant tissues. RICTOR amplification was tested by Droplet Digital PCR (ddPCR) and validated using the "gold standard" fluorescence in situ hybridisation (FISH). Additionally, the consequences of Rictor protein expression were also studied by immunohistochemistry. RICTOR amplification was presumed in 37 cases with CNV ≥ 3 by NGS, among these, 16 cases (16/420; 3.8%) could be validated by FISH, however, ddPCR confirmed only 11 RICTOR-amplified cases with lower sensitivity. Based on these, neither NGS nor ddPCR could replace traditional FISH in proof of RICTOR amplification. However, NGS could be beneficial to highlight potential RICTOR-amplified cases. The obtained results of the 14 different tumour types with FISH-validated RICTOR amplification demonstrate the importance of RICTOR amplification in a broad spectrum of tumours. The newly described RICTOR-amplified entities could initiate further collaborative studies with larger cohorts to analyse the prevalence of RICTOR amplification in rare diseases. Finally, our and further work could help to improve and expand future therapeutic opportunities for mTOR-targeted therapies.
Subject(s)
DNA Copy Number Variations , Neoplasms , Humans , Neoplasms/genetics , TOR Serine-Threonine Kinases/genetics , Rapamycin-Insensitive Companion of mTOR Protein/genetics , High-Throughput Nucleotide Sequencing , Gene AmplificationABSTRACT
Significant improvements in the survival rates of paediatric cancer have been achieved over the past decade owing to recent advances in therapeutic and diagnostic strategies. However, disease progression and relapse remain a major challenge for the clinical management of paediatric angiosarcoma. Comprehensive genomic profiling of these rare tumours using high-throughput sequencing technologies may improve patient stratification and identify actionable biomarkers for therapeutic intervention. Here, we describe the clinical, histopathological, immunohistochemical and molecular profile of a novel and precision medicine-informed case where a KHDRBS1-NTRK3 fusion determined by next-generation sequencing-based comprehensive genomic profiling led to complete and sustained remission (clinical and radiological response) in an otherwise incurable disease. Our patient represents the first paediatric angiosarcoma harbouring a targetable NTRK3 fusion in the literature and demonstrates the first example of targeting this alteration in angiosarcoma using larotrectinib, an NTRK inhibitor. Clinical and radiological remission was achieved in under two months of therapy, and the patient is currently in complete remission, 4 month after stopping larotrectinib therapy, which was given over 17 months with only mild side effects reported. Therefore, this remarkable case exemplifies the true essence of precision-based care by incorporating conventional pathology with the why, when, and how to test for rare oncogenic drivers and agnostic biomarkers in paediatric angiosarcoma.
ABSTRACT
Due to the relatively high recurrence rate and the destructive nature of the tumor, the treatment of giant cell tumor is still a challenge. Denosumab appeared to be a promising candidate as a therapeutic drug. However, several studies have reported that tumors can recur during/after treatment with denosumab. Based on activated receptor tyrosine kinase signaling pattern of the stromal/tumor cells, a combination treatment with denosumab and sunitinib has recently been proposed to inhibit recurrences. This prompted us to investigate the PDGFRß expression of five denosumab treated cases using both primary and recurrent tumors during and after denosumab treatment. In addition, to recognise morphological changes, immunohistochemical analysis of H3F3A and PDGFRß was also performed. As an effect of denosumab treatment, the permanent absence of giant cells associated with severe to mild fibrosis was the most consistent morphological change, but H3F3A positive stromal/tumor cells were observed in all cases. Furthermore, an increased immunopositivity of PDGFRß in stromal/tumor cells was evident in all recurrent cases during denosumab treatment. Upon tumor recurrence (after the discontinuation of denosumab treatment) the intensity of PDGFRß immunostaining in stromal/tumor cells was restored/decreased. Our results confirm (for the first time) the activation of PDGFRß on mononuclear stromal/tumor cells at protein level as an effect of denosumab treatment, which has so far only been demonstrated by phosphoprotein array analysis (protein lysates). The decreased PDGFRß activity after the discontinuation of denosumab treatmeant and the increased PDGFRß activity during denosumab treatment underlines the need for denosumab and sunitinib combination therapy.
Subject(s)
Bone Neoplasms , Giant Cell Tumor of Bone , Soft Tissue Neoplasms , Bone Neoplasms/pathology , Bone and Bones/pathology , Denosumab/therapeutic use , Giant Cell Tumor of Bone/drug therapy , Giant Cell Tumor of Bone/pathology , Humans , Sunitinib/therapeutic useABSTRACT
Background: Chordoma, the most frequent malignant primary spinal neoplasm, characterized by a high rate of recurrence, is an orphan disease where the clarification of the molecular oncogenesis would be crucial to developing new, effective therapies. Dysregulated expression of non-coding RNAs, especially microRNAs (miRNA) has a significant role in cancer development. Methods: Next-generation RNA sequencing (NGS) was used for the combinatorial analysis of mRNA-miRNA gene expression profiles in sacral chordoma and nucleus pulposus samples. Advanced bioinformatics workflow was applied to the data to predict miRNA-mRNA regulatory networks with altered activity in chordoma. Results: A large set of significantly dysregulated miRNAs in chordoma and their differentially expressed target genes have been identified. Several molecular pathways related to tumorigenesis and the modulation of the immune system are predicted to be dysregulated due to aberrant miRNA expression in chordoma. We identified a gene set including key regulators of the Hippo pathway, which is targeted by differently expressed miRNAs, and validated their altered expression by RT-qPCR. These newly identified miRNA/RNA interactions are predicted to have a role in the self-renewal process of chordoma stem cells, which might sustain the high rate of recurrence for this tumor. Conclusions: Our results can significantly contribute to the designation of possible targets for the development of anti-chordoma therapies.
ABSTRACT
BACKGROUND Wilms' tumor is a common renal malignancy of early childhood with a generally favorable prognosis depending upon histological subtype. It is becoming increasingly clear that differences in miRNA (microRNA) expression signature represent important clues helping us predict a tumor's response to chemotherapy. In our study, we aimed to reveal miRNAs deregulated in regressive Wilms' tumors from FFPE (formalin-fixed, paraffin-embedded) samples, also showing whether such samples are reliable miRNA sources in Wilms' tumor. MATERIAL AND METHODS Samples from 8 Hungarian patients (3 males, 5 females, aged 1 to 7 years) were analyzed by qRT-PCR (quantitative real-time PCR). A PCR array was used in a pilot experiment, and selected miRNAs (miR-128-3p, miR-184, miR-194-5p, miR-203a) were studied in the rest of the samples using individual primers. RESULTS miR-194-5p was underexpressed in all tumor samples. miR-184 and miR-203a were underexpressed in 7 cases, the exception being a case with a high ratio of necrotic blastemal tissue. Results obtained with miR-128-3p are difficult to interpret due to varying directions of expression changes. CONCLUSIONS We conclude that a downregulation of miR-184, miR-194-5p, and miR-203a expression is observed in both regressive and blastemal tumors, but larger-scale studies are needed to confirm whether the degree of their underexpression correlates with the number of blastemal elements in a sample. In most of our FFPE samples aged up to 9 years, RNA extraction provided miRNA with quantity and quality sufficient for qRT-PCR-based analysis, emphasizing the relevance of pathological archives as miRNA sources in future studies.
Subject(s)
Gene Expression Profiling/methods , Kidney Neoplasms/genetics , MicroRNAs/genetics , Paraffin Embedding/methods , Real-Time Polymerase Chain Reaction/methods , Wilms Tumor/genetics , Child , Child, Preschool , Female , Gene Expression Regulation, Neoplastic , Humans , Hungary , Infant , MaleABSTRACT
The self-limited nature of nodular fasciitis (NF) is well-known but its precise mechanism has not yet been clarified. We observed that "young" NF (preoperative duration <1 month) consistently contains a higher percentage (~80%) of USP6 break-apart FISH signals than "old" NF (preoperative duration >3 months) (~20%). Thus, we hypothesized that our original observation may reflect a connection with the self-limited nature of NF. Seventeen cases with reliable data concerning the onset were selected, thus approximating the lifetime of each tumor. Besides the USP6 interphase FISH examination, we also checked the most common MYH9-USP6 fusion using RT-PCR. Because of the known pathways of the tumorigenesis of NF, the mRNA level of USP6, TRAIL, IFN-beta, JAK1, STAT1, STAT3, JUN, and CDKN2A was measured using qRT-PCR. Regarding proteins, USP6, p16, p27, TRAIL, and IFN-beta were examined using immunohistochemistry. Targeted gene panel next-generation sequencing (NGS) of three cases was additionally performed. We found a strong negative correlation (p = 0.000) between the lifetime and percentage of USP6 break-apart signals and a strong positive relationship (p = 0.000) between USP6 break-apart signals and mitotic counts. Results of immunostainings, along with qRT-PCR results, favored the previously-suggested USP6-induced negative feedback mechanism through activation of TRAIL and IFN-beta, likely resulting in apoptosis and senescence of tumor cells harboring USP6 fusions. Targeted-NGS resulted in the detection of several variants, but no additional recurrent changes in the pathogenesis of these tumors. We revealed on a cellular level the USP6-induced negative feedback mechanism. In conclusion, we emphasize that in "old" NF, the percentage of USP6 break-apart FISH signals can be as low as 14-27% which can be very important from a differential diagnostic point of view. We emphasize that a careful examination and interpretation of the NGS data is needed before clinical decision-making on treatment.
Subject(s)
Biomarkers, Tumor/genetics , Fasciitis/genetics , Gene Fusion , Gene Rearrangement , Neoplasms, Connective Tissue/genetics , Soft Tissue Neoplasms/genetics , Ubiquitin Thiolesterase/genetics , Adolescent , Adult , Biomarkers, Tumor/analysis , Child , Child, Preschool , Fasciitis/metabolism , Fasciitis/pathology , Female , High-Throughput Nucleotide Sequencing , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Male , Middle Aged , Myofibroblasts/chemistry , Myofibroblasts/pathology , Neoplasms, Connective Tissue/chemistry , Neoplasms, Connective Tissue/pathology , Real-Time Polymerase Chain Reaction , Reverse Transcriptase Polymerase Chain Reaction , Soft Tissue Neoplasms/chemistry , Soft Tissue Neoplasms/pathology , Time Factors , Young AdultABSTRACT
Cells of the monocyte macrophage lineage form multinucleated giant cells (GCs) by fusion, which may express some cell cycle markers. By using a comprehensive marker set, here we looked for potential replication activities in GCs, and investigated whether these have diagnostic or clinical relevance in giant cell tumor of bone (GCTB). GC rich regions of 10 primary and 10 first recurrence GCTB cases were tested using immunohistochemistry in tissue microarrays. The nuclear positivity rate of the general proliferation marker, replication licensing, G1/S-phase, S/G2/M-phase, mitosis promoter, and cyclin dependent kinase (CDK) inhibitor reactions was analyzed in GCs. Concerning Ki67, moderate SP6 reaction was seen in many GC nuclei, while B56 and Mib1 positivity was rare, but the latter could be linked to more aggressive (p = 0.012) phenotype. Regular MCM6 reaction, as opposed to uncommon MCM2, suggested an initial DNA unwinding. Early replication course in GCs was also supported by widely detecting CDK4 and cyclin E, for the first time, and confirming cyclin D1 upregulation. However, post-G1-phase markers CDK2, cyclin A, geminin, topoisomerase-2a, aurora kinase A, and phospho-histone H3 were rare or missing. These were likely silenced by upregulated CDK inhibitors p15INK4b, p16INK4a, p27KIP1, p53 through its effector p21WAF1 and possibly cyclin G1, consistent with the prevention of DNA replication. In conclusion, the upregulation of known and several novel cell cycle progression markers detected here clearly verify early replication activities in GCs, which are controlled by cell cycle arresting CDK inhibitors at G1 phase, and support the functional maturation of GCs in GCTB.
Subject(s)
Biomarkers, Tumor/metabolism , Bone Neoplasms/pathology , Cell Cycle Proteins/metabolism , Cell Cycle , Cell Proliferation , Giant Cell Tumor of Bone/pathology , Adolescent , Adult , Aged , Bone Neoplasms/metabolism , Female , Follow-Up Studies , Giant Cell Tumor of Bone/metabolism , Humans , Male , Middle Aged , Prognosis , Retrospective Studies , Tumor Cells, Cultured , Young AdultSubject(s)
Neoplasms , Receptor, trkA/genetics , Gene Fusion , Humans , Neoplasms/diagnosis , Neoplasms/geneticsABSTRACT
Although papillary endothelial hyperplasia may occur at almost any site, one of the most common sites is the hand. It is generally regarded as a reactive vascular proliferation i.e. exuberant form of organizing thrombus. Diagnosis of Masson tumor can be challenging due to its close clinical, radiological and even histopathological resemblance to angiosarcoma. We present seven cases of Masson tumor of the hand; wanting to reveal its nature using new vascular markers and discuss the treatment options and expected outcomes, present clinical and radiological features that may aid diagnosis and also offer treatment plans. A multicenter retrospective study was performed between January 2014 and November 2019. Immunohistochemical stains of Glut1, WT1, ERG, CD31 and alpha smooth muscle actin (ASMA) were performed on each cases. We found seven cases during the examined period. 4 out of 7 cases were women. All lesions occurred in the hands. 3 out of 7 cases appeared in a previously present vascular malformation. All cases were treated with surgical excision and the diagnosis of papillary endothelial hyperplasia was made by histology. Pre-operative testing (radiograph/MRI/US/fine needle aspiration biopsy) did not suggest the diagnosis of Masson tumor; however, aspiration cytology could rule out malignancy. The proliferative endothelial cells proved to be Glut1 negative and WT1 positive and the accompanying pericytic cells were ASMA positive in all cases. Though Masson tumor is a rare vascular lesion in the hand among other vascular tumors, it should be considered in the differential diagnostics even in the case of previously existing vascular malformation. WT1 positivity of the endothelial cells and the accompanying pericytic cells raises the question whether the initially reactive endothelial proliferation may transform into a true benign vascular tumor.
Subject(s)
Biomarkers/analysis , Carcinoma, Papillary/pathology , Endothelium, Vascular/pathology , Hand/pathology , Hyperplasia/pathology , Vascular Neoplasms/pathology , Adult , Aged , Biomarkers/metabolism , Carcinoma, Papillary/metabolism , Carcinoma, Papillary/surgery , Endothelium, Vascular/metabolism , Endothelium, Vascular/surgery , Female , Follow-Up Studies , Hand/surgery , Humans , Hyperplasia/metabolism , Hyperplasia/surgery , Male , Middle Aged , Prognosis , Retrospective Studies , Vascular Neoplasms/metabolism , Vascular Neoplasms/surgeryABSTRACT
mTOR activation has been observed in rhabdomyosarcoma (RMS); however, mTOR complex (mTORC) 1 inhibition has had limited success thus far. mTOR activation alters the metabolic pathways, which is linked to survival and metastasis. These pathways have not been thoroughly analyzed in RMSs. We performed immunohistochemistry on 65 samples to analyze the expression of mTOR complexes (pmTOR, pS6, Rictor), and several metabolic enzymes (phosphofructokinase, lactate dehydrogenase-A, ß-F1-ATPase, glucose-6-phosphate dehydrogenase, glutaminase). RICTOR amplification, as a potential mechanism of Rictor overexpression, was analyzed by FISH and digital droplet PCR. In total, 64% of the studied primary samples showed mTOR activity with an mTORC2 dominance (82%). Chemotherapy did not cause any relevant change in mTOR activity. Elevated mTOR activity was associated with a worse prognosis in relapsed cases. RICTOR amplification was not confirmed in any of the cases. Our findings suggest the importance of the Warburg effect and the pentose-phosphate pathway beside a glutamine demand in RMS cells. The expression pattern of the studied mTOR markers can explain the inefficacy of mTORC1 inhibitor therapy. Therefore, we suggest performing a detailed investigation of the mTOR profile before administering mTORC1 inhibitor therapy. Furthermore, our findings highlight that targeting the metabolic plasticity could be an alternative therapeutic approach.
ABSTRACT
Pulmonary arterial intimal sarcoma is a rare tumour with high mortality. Due to its localisation, the symptoms can mimic pulmonary thromboembolism and pneumonia, therefore assessing the diagnosis and choosing the adequate therapy is never easy. Its therapy mainly consists of surgery combined with radiochemotherapy. A 46-year-old male patient with testicular seminoma, pulmonary embolism, bronchial asthma and pollen allergy in his history had a follow-up thoracic CT. On the left side of the lung, a pleura-infiltrating 7-8 cm lesion, which occluded the pulmonary artery, was described. The first biopsy specimen showed fragments of spindle cell tumour. The primary diagnosis was leiomyoma, leiomyomatous hyperplasia, yet the presence of leiomyosarcoma could not have been ruled out. Due to the arterial obstruction, the patient underwent left sided pulmonectomy. Histological examination showed a tumour mostly composed of spindle cells that were diffusely positive with SMA, focally diffuse with MDM2 immunohistochemistry together with high proliferation activity. h-Caldesmon, S-100, ERG and pancytokeratin expressions were not detected. With fluorescent in situ hybridization 10% of tumour cells showed polysomy and MDM2 amplification. According to the results, high-grade pulmonary arterial intimal sarcoma diagnosis has been made. The precise incidence of pulmonary arterial intimal sarcoma is unknown. Some literature data suggest it can be the cause of chronic pulmonary hypertension in 1-4% of the cases. Weight loss can draw attention to the malignant nature of the disease. Imaging techniques and histology are the gold standard in setting the diagnosis. The prognosis is poor. Early recognition and surgery combined with chemotherapy can prolong survival. Orv Hetil. 2020; 161(6): 232-236.
Subject(s)
Pulmonary Artery , Sarcoma/diagnosis , Vascular Neoplasms/diagnosis , Humans , In Situ Hybridization, Fluorescence , Male , Middle AgedABSTRACT
Thirty-one cases of low-grade renal cell carcinoma (RCC) with clear cells and tubulopapillary/papillary architecture were analyzed retrospectively with immunohistochemical and genetic markers to gain more experience with the differential diagnosis of such cases. All samples coexpressed CK7 and CA9; the TFE3 or TFEB reactions were negative; the CD10 and the AMACR stainings were negative in 27 cases and 30 cases, respectively. The FISH assays for papillary RCC, available in 27 cases, and deletion of chromosome 3p, available in 29 cases, gave negative results. The results for 3p deletion, VHL gene mutation or VHL gene promoter region hypermethylation testing, along with the diffuse CD10-positivity in 2 cases confirmed 21 cases as clear cell papillary RCC (CCPRCC; CK7+, CA9+; no 3p loss, no VHL abnormality) and 10 cases as clear cell RCC (CCRCC; CK7+, CA9+; no 3p loss, VHL abnormality mutation/hypermethylation present). In CCPRCCs, the representative growth pattern was branching tubulo-acinar, commonly accompanied by cyst formation. The linear nuclear arrangement or cup-shaped staining of CA9 did not necessarily indicate CCPRCC, and the absence of these did not exclude the diagnosis of CCPPRC. One tumor infiltrated the renal sinus; the others exhibited pT1 stage; and metastatic outcome was not recorded. The CCRCC cases were in pT1 stage; 6 exhibited cup-shaped staining of CA9, and 1 displayed lymph node metastasis at the time of surgery. Distant metastatic disease was not observed. In summary, the VHL abnormalities distinguished the subset of CCRCC with diffuse CK7-positivity and no 3p loss from cases of CCPRCC.
Subject(s)
Biomarkers, Tumor/analysis , Carcinoma, Papillary/diagnosis , Carcinoma, Renal Cell/diagnosis , Kidney Neoplasms/diagnosis , Adult , Aged , Aged, 80 and over , Carcinoma, Papillary/genetics , Carcinoma, Renal Cell/genetics , DNA Methylation/genetics , Diagnosis, Differential , Female , Humans , Kidney Neoplasms/genetics , Male , Middle Aged , Retrospective Studies , Von Hippel-Lindau Tumor Suppressor Protein/geneticsABSTRACT
Coincidences of more than one pathogenic mutation in high and/or moderate risk-associated cancer genes have been rarely reported, and the implication for disease progression has been debated. We present a case harboring two autosomal dominant inherited mutations potentially aggravating the phenotype. Case report: A 16-year-old female was referred to the Endocrine Unit due to two palpable thyroid nodules and hair loss. Two hypoechoic, inhomogeneous masses with microcalcification in the thyroid gland were confirmed as medullary thyroid carcinoma. Genetic testing revealed a pathogenic heterozygous RET mutation associated with multiple endocrine neoplasia type 2 (MEN2). Furthermore, genetic screening identified the same mutation in the proband's clinically negative brother as well as in his two sons. The proband's mother and maternal aunt died of breast cancer. No samples were available from the deceased. The proband underwent further genetic counseling and BRCA1/2 testing. A novel, frameshift heterozygous BRCA1 mutation (BRCA1 p.Ile90Serfs, NC_000017.10:g.41256905_41256917) was identified in the proband, but it was absent in the brother and father, indicative of maternal inheritance. Breast or ovarian cancer was neither detected in our case at initial presentation nor during the 6-year follow-up. Conclusion: Coincidence of two monogenic autosomal dominant tumor syndromes is extremely rare, but it represents a significant therapeutic and cancer surveillance challenge. Due to the wider use of next generation sequencing in clinical practice, similar situations may occur more frequently.
ABSTRACT
OBJECTIVE: Giant cell tumor of bone (GCTB) is a frequently recurring locally aggressive osteolytic lesion, where pathological osteoclastogenesis and bone destruction are driven by neoplastic stromal cells. Here, we studied if cell cycle fractions within the mononuclear cell compartment of GCTB can predict its progression-free survival (PFS). METHODS: 154 cases (100 primaries and 54 recurrent) from 139 patients of 40 progression events, was studied using tissue microarrays. Ploidy and in situ cell cycle progression related proteins including Ki67 and those linked with replication licensing (mcm2), G1-phase (cyclin D1, Cdk4), and S-G2-M-phase (cyclin A; Cdk2) fractions; cell cycle control (p21waf1) and repression (geminin), were tested. The Prentice-Williams-Peterson (PWP) gap-time models with the Akaike information criterion (AIC) were used for PFS analysis. RESULTS: Cluster analysis showed good correlation between functionally related marker positive cell fractions indicating no major cell cycle arrested cell populations in GCTB. Increasing hazard of progression was statistically associated with the elevated post-G1/S-phase cell fractions. Univariate analysis revealed significant negative association of poly-/aneuploidy (pâ¯<â¯0.0001), and elevated cyclin A (pâ¯<â¯0.001), geminin (pâ¯=â¯0.015), mcm2 (pâ¯=â¯0.016), cyclin D1 (pâ¯=â¯0.022) and Ki67 (B56: pâ¯=â¯0.0543; and Mib1: pâ¯=â¯0.0564 -strong trend) positive cell fractions with PFS. The highest-ranked multivariate interaction model (AICâ¯=â¯269.5) also included ploidy (HR 5.68, 95%CI: 2.62-12.31, pâ¯<â¯0.0001), mcm2 (pâ¯=â¯0.609), cyclin D1 (HR 1.89, 95%CI: 0.88-4.09, pâ¯=â¯0.105) and cyclin A (pâ¯<â¯0.0001). The first and second best prognostic models without interaction (AICâ¯=â¯271.6) and the sensitivity analysis (AICâ¯=â¯265.7) further confirmed the prognostic relevance of combining these markers. CONCLUSION: Ploidy and elevated replication licensing (mcm2), G1-phase (cyclin D1) and post-G1 phase (cyclin A) marker positive cell fractions, indicating enhanced cell cycle progression, can assist in identifying GCTB patients with increased risk for a reduced PFS.
Subject(s)
Cell Cycle , Giant Cell Tumor of Bone/pathology , Adolescent , Adult , Aged , Biomarkers, Tumor/metabolism , Child , Child, Preschool , Cohort Studies , Disease Progression , Female , Humans , Kaplan-Meier Estimate , Ki-67 Antigen/metabolism , Male , Middle Aged , Models, Biological , Multivariate Analysis , Progression-Free Survival , Risk Factors , Young AdultABSTRACT
Blastemal Wilms' tumors are associated with poor chemo-responsiveness and an adverse prognosis. Our aim was to contribute to the miRNA profiling of the disease, while demonstrating the value of archived formalin-fixed, paraffin-embedded (FFPE) samples as miRNA sources. MiRNA was extracted from tumor and normal tissues of 8 patients diagnosed with blastemal Wilms' tumor in Hungary. A quantitative real-time PCR-based protocol was used to identify miRNAs of interest and study the expression of selected miRNAs in all samples. Profiling of miRNA expression from FFPE samples turned out to be cost-effective in Wilms' tumor, as most miRNAs (including miRNA-194-5p, which was studied in all patients) showed expression alterations similar to the ones reported in the literature. MiR-184 expression was found to be lower than in previous studies, while the downregulation of miR-203a is a novel finding. MiR-184 may be downregulated in a subset of blastemal and other Wilms' tumors. A loss of miR-203a may or may not be specific to blastemal cells, but available evidence hints at its importance in the pathogenesis of Wilms' tumor. It should be considered for inclusion in future studies of miRNA expression.
Subject(s)
MicroRNAs/genetics , Wilms Tumor/genetics , Child , Child, Preschool , Female , Gene Expression Regulation, Neoplastic/genetics , Humans , Infant , Male , Paraffin Embedding , Prognosis , Reverse Transcriptase Polymerase Chain Reaction , Wilms Tumor/pathologyABSTRACT
Soft tissue sarcomas (STS) and neuroblastomas (NBL), are childhood malignancies still associated with poor prognoses despite the overall improvement in childhood tumor survival of the past decades. Anaplastic lymphoma kinase (ALK) inhibition is promising new strategy to improve the outcome of these pediatric tumors. Eighteen histologic samples of pediatric STS and 19 NBL patients were analyzed for ALK abnormalities using fluorescent in situ hybridization (FISH) with break-apart probes and immunohistochemistry (IHC). ALK alterations were presented in 20 of the 37 sections. The presence of ALK alteration in NBL samples were detected using IHC in 84,2% of all cases compared to 21,1% FISH positivity. In STS cases the results were less different (IHC 16,7% vs FISH 22,2%). The difference can be explained by the different type of molecular alterations. FISH method detected translocation and amplification, but not the point mutation of ALK gene. IHC confirmed the diagnosis by detecting the expression of ALK protein.After ALK positivity was proven, the effectiveness and safety of the crizotinib therapy was examined in 4 patients (1 alveolar rhabdomyosarcoma (RMA), 1 embryonal rhabdomyosarcoma (RME), 1 inflammatory myofibroblastic tumor (IMT), 1 NBL). We observed continuous remission of the IMT patient, all other cases the inhibitor treatment was not curative.Our findings underline the importance of screening the ALK status parallel with both IHC and FISH. Crizotinib treatment had a long-term effect in ALK positive IMT patients, however itwas only temporary efficient in relapsed, progressive STS and NBL.
Subject(s)
Anaplastic Lymphoma Kinase/genetics , Crizotinib/therapeutic use , Inflammation/genetics , Neoplasms, Muscle Tissue/genetics , Rhabdomyosarcoma, Alveolar/genetics , Rhabdomyosarcoma, Embryonal/genetics , Adolescent , Anaplastic Lymphoma Kinase/metabolism , Biomarkers, Tumor/genetics , Child , Child, Preschool , Female , Follow-Up Studies , Gene Amplification , Humans , Immunohistochemistry , In Situ Hybridization, Fluorescence , Infant , Infant, Newborn , Inflammation/drug therapy , Inflammation/pathology , Male , Neoplasms, Muscle Tissue/drug therapy , Neoplasms, Muscle Tissue/pathology , Point Mutation , Prognosis , Protein Kinase Inhibitors/therapeutic use , Rhabdomyosarcoma, Alveolar/drug therapy , Rhabdomyosarcoma, Alveolar/pathology , Rhabdomyosarcoma, Embryonal/drug therapy , Rhabdomyosarcoma, Embryonal/pathology , Translocation, GeneticABSTRACT
The survival of children treated with Ewing sarcoma at Semmelweis University were investigated. Pediatric patients with Ewing sarcoma treated at Semmelweis University from 2001 through 2013 were analyzed in terms of overall survival and clinical factors (age, primary localization and extent of the tumor, time interval from primary complaints to diagnosis). For statistical analysis Kaplan-Meier estimated survival and log rank test were applied. Mean age and follow-up time of the 78 patients were 11.16 and 6.29 years, respectively. In 57% of patients time interval from primary symptoms to diagnosis was less than half year. In 53.8% of the patients the disease was metastatic at primary diagnosis (pulmonary only: 29.5%, any other: 24.3%). 5- and 10-year overall survival of patients were 68.1% and 60.4%, respectively. Among the analyzed factors, the presence of metastasis impaired 5-year overall survival significantly (88.5% for localized disease, 63.5% for pulmonary only and 40.9% for any other metastasis). The survival rate of pediatric patients with Ewing sarcoma treated at Semmelweis University is similar to the result in Western European countries.