Search

Background Mantle cell lymphoma (MCL) is derived from naïve CD5+ B-cells with the cytogenetic hallmark translocation 11;14. The presence of additional abnormalities is associated with blastoid variants in MCL (BMCL) and confers a poor prognosis. Many of these tumors also show deletion or loss of heterozygosity (LOH) of the ATM gene and biallelic ATM inactivation show significantly higher chromosomal imbalances. Case presentation Here we report a 52 year-old male who presented to the clinic with worsening dyspnea, fever, chills, diffuse lymphadenopathy, splenomegaly and leukocytosis with blastoid cells circulating in blood. The bone marrow aspirate showed about 40% abnormal blast-looking cells and biopsy revealed a remarkable lymphoid infiltrate. The patient was diagnosed with blastoid variant mantle cell lymphoma (BMCL). Chromosome analysis on bone marrow showed a complex karyotype. FISH analysis from B-cell lymphoma panel showed bi-allelic amplification of ATM gene. Other abnormalities were present including CCND1/IGH fusion, confirming the MCL diagnosis, in addition to RB1 and p53 deletion. High resolution SNP-microarray studies showed complex copy number changes, especially on chromosomes 7 and 11, consistent with chromoanagenesis. Microarray studies also showed LOH at the ATM locus indicating the amplification seen on FISH is not biallelic. Conclusion To the best of our knowledge, ATM gene amplification is not previously reported in BMCL and our case suggests a novel mechanism of ATM inactivation caused by chromoanagenesis resulting in mutant allele specific imbalance with copy number gain.

Cat scratch disease (CSD) is an infectious disease process of generally immunocompetent children and young adults. This infection can be introduced through skin trauma by direct exposure to the saliva of an infected kitten or cat. CSD is typically associated with constitutional symptoms and self-limited regional lymphadenopathy. In the sole presence of swollen lymph nodes, however, the differential diagnosis for CSD is relatively broad, including an active infection, an ongoing inflammatory process, and a metastatic process. CSD can present as axillary lymphadenopathy without typical constitutional symptoms. With proper clinical and laboratory investigation, CSD can be accurately identified and correctly diagnosed, as demonstrated in this case series featuring five symptomatic young adults with axillary lymphadenopathy. Breast imaging clinic specializes in lymph node assessment because metastatic lymphadenopathy is one of the most common presenting signs of breast cancer. Most isolated axillary lymphadenopathy without breast mass is benign reactive lymphadenopathy, but biopsy is necessary to exclude malignancies, such as metastatic lymphadenopathy or lymphoma.

Identification of the functional impact of mutated and altered genes in cancer is critical for implementing precision oncology and drug repurposing. In recent years, the emergence of multiomics data from large, well-characterized patient cohorts has provided us with an unprecedented opportunity to address this problem. In this study, we investigated survival-associated genes across 26 cancer types and found that these genes tended to be hub genes and had higher K-core values in biological networks. Moreover, the genes associated with adverse outcomes were mainly enriched in pathways related to genetic information processing and cellular processes, while the genes with favorable outcomes were enriched in metabolism and immune regulation pathways. We proposed using the number of survival-related neighbors to assess the impact of mutations. In addition, by integrating other databases including the Human Protein Atlas and the DrugBank database, we predicted novel targets and anticancer drugs using the drug repurposing strategy. Our results illustrated the significance of multidimensional analysis of clinical data in important gene identification and drug development.

Hepatocellular adenoma (HCA) is a benign hepatocyte-derived tumor commonly seen in reproductive-aged women with long-term use of oral contraceptives (OCs) in European and North American countries. Accordingly, HCA is currently classified into four molecular subtypes as adopted by the World Health Organization. The present study was firstly to characterize and determine the genetic alterations and clinicopathological features of the largest series of HCAs in China. We reviewed 189 patients with HCA who underwent hepatectomies at our liver center from January 1984 to January 2012, among which 36 HCAs were randomly selected for the sequencing of HNF1α, β-catenin and gp130 genes, and 60 HCAs were randomly selected for detecting microsatellite instability (MSI). Compared with Western studies, our data showed distinctive findings including male (69.8%) and overweight/obese (50.3%) predominance. Only 3.5% of female patients had a documented history of OCs use for 2-4 years. All 36 sequenced HCAs showed HNF1α mutations (72% missense, 28% synonymous), 2 hotspot polymorphisms of HNF1α (I27L: rs1169288 and S487N: rs2464196) were seen in 17 (47%) and 10 (27.8%) cases, respectively, and a novel single nucleotide polymorphism site (rs1169304) in intron 9 of HNF1α was detected in 32 (88%) cases, but no β-catenin or gp130 gene mutation was detected, and no nuclear β-catenin staining was detected by immunohistochemistry. The frequency of MSI was 75% for D12S1398 (HNF1α inactivated pathway) and 78.5% for D6S1064 (HIPPO signaling pathway) in 34 overweight/obese patients with HCA. Our results firstly indicate that patients with HCA in China frequently occur in male overweigh and obese adult population, lack an association with OCs use and exhibit unique genetic alterations. Taken together, these observations suggest that alternative pathogenetic pathways involve in HCA tumorigenesis in Chinese patients.

We report a case of a 43-year-old male who presented with a large soft-tissue neck mass 7 years ago. A diagnosis of giant cell angiofibroma (GCA) was made on the basis of light microscopy and immunohistochemical studies. Chromosome analysis showed a male karyotype with t(12;17)(q15;q23),del(18)(q21) in all 20 cells analyzed. This cytogenetic abnormality in GCA is different from the t(17;22)(q22;q13) found in related lesions, such as giant cell fibroblastoma and solitary fibrous tumor, none of which has a specific chromosomal abnormality. Our case is the second case of GCA with chromosomal aberrations. To the best of our knowledge, this is the first case of GCA with t(12;17) occurring as an extra-orbital mass.

Pro-tumorigenic function of the p38 kinase plays a critical role in human cholangiocarcinogenesis. However, the underlying mechanism remains incompletely understood. Here, we report that c-Met, the tyrosine kinase receptor for hepatocyte growth factor (HGF), contributes to the pro-tumorigenic ability of p38 in human cholangiocarcinoma cells. Both p38 and c-Met promote the proliferation and invasion of human cholangiocarcinoma cells. Importantly, inhibition or knockdown of p38 decreased the basal activation of c-Met. Tyrosine phosphatase inhibitor studies revealed that p38 promotes the activity of c-Met, at least in part, by inhibiting dephosphorylation of the receptor. Moreover, density enhanced phosphatase-1 (DEP-1) is involved in p38-mediated inhibiting dephosphorylation of c-Met. Furthermore, p38 inhibits the degradation of c-Met. Taken together, these data provide a potential mechanism to explain how p38 promotes human cholangiocarcinoma cell proliferation and invasion. We propose that the link between p38 and c-Met is implicated in the progression of human cholangiocarcinoma.

Acute promyelocytic leukemia (APL) is a well-defined subtype of acute myeloid leukemia (AML) specifically characterized by the t(15;17)(q22;q12) translocation. The t(15;17) results in the fusion of the promyelocytic leukemia (PML) and retinoic acid receptor alpha (RARA) genes. Rare cryptic fusions often associated with small genomic insertions can best be detected by reverse transcriptase-polymerase chain reaction (RT-PCR) although conventional chromosomal studies or even fluorescence in situ hybridization (FISH) analyses appear normal. We report here an APL clone with a cryptic PML-RARA fusion that returned negative results by both karyotyping and fluorescence in situ hybridization (FISH), but returned positive results by RT-PCR analysis. A single nucleotide polymorphism (SNP) microarray analysis was used in this case to help resolve the discordance, revealing a 49-kilobase intragenic PML gene duplication. A dual color dual fusion PML-RARA FISH probe set identified a small, extra PML signal in a chromosome other than 15 or 17. Although coinsertion of a RARA sequence could be detected by neither FISH nor array, the RT-PCR positivity is consistent with this fusion "ectopic" to the natural gene loci. The findings highlight the clinical utility of microarray in cases of cryptic PML-RARA fusion.

c-Met, the tyrosine-kinase receptor for hepatocyte growth factor, plays a critical role in the tumorigenesis of hepatocellular carcinoma (HCC). However, the underlying mechanism remains incompletely understood. The mature c-Met protein p190Met(αβ) (consists of a α subunit and a β subunit) is processed from pro-Met. Here we show that pro-Met is processed into p190Met(NC) by sarco/endoplasmic reticulum calcium-ATPase (SERCA) inhibitor thapsigargin. p190Met(NC) compensates for the degradation of p190Met(αβ) and protects human HCC cells from apoptosis mediated by endoplasmic reticulum (ER) stress. In comparison with p190Met(αβ), p190Met(NC) is not cleaved and is expressed as a single-chain polypeptide. Thapsigargin-initiated p190Met(NC) expression depends on the disturbance of ER calcium homeostasis. Once induced, p190Met(NC) is activated independent of hepatocyte growth factor engagement. p190Met(NC) contributes to sustained high basal activation of c-Met downstream pathways during ER calcium disturbance-mediated ER stress. Both p38 MAPK-promoted glucose-regulated protein 78 (GRP78) expression and sustained high basal activation of PI3K/Akt and MEK/ERK are involved in the cytoprotective function of p190Met(NC). Importantly, the expression of p190Met(NC) is detected in some HCC cases. Taken together, these data provide a potential mechanism to explain how c-Met promotes HCC cells survival in response to ER stress. We propose that context-specific processing of c-Met protein is implicated in HCC progression in stressful microenvironments.

Background & Aims Hepatocellular carcinoma (HCC) is believed to arise from tumor-initiating cells (T-ICs), although little is known about their stem cell–like properties. Methods We quantified levels of p28 GANK (Gankyrin), OV6, and Oct4 in 130 human HCC samples using immunohistochemistry. Magnetic-activated cell sorting was used to isolate OV6 + HCC cells. T-IC properties were evaluated by quantitative reverse-transcription polymerase chain reaction, flow cytometry, and spheroid formation. We used a coimmunoprecipitation assay to study interactions among p28 GANK , Oct4, and WWP2. Tumorigenicity and pulmonary metastasis were examined in nonobese diabetic and severe combined immunodeficient mice. Results In HCC samples, high levels of p28 GANK correlated with expansion of OV6 + tumor cells; the combination of high levels of p28 GANK and OV6 was associated with progression of HCC. p28 GANK was predominantly expressed in liver T-ICs, isolated by magnetic sorting, and undifferentiated primary HCC spheroids. Increased levels of p28 GANK in T-ICs increased their percentages in HCC samples, expression of stem cell genes, self-renewal potential, chemoresistance in vitro, and tumorigenicity and ability to develop into pulmonary metastases in mice. Conversely, knockdown of p28 GANK reduced their T-IC properties. p28 GANK likely activates liver T-ICs by impeding ubiquitination and degradation of the transcription factor Oct4 by WWP2. In support of this concept, levels of p28 GANK correlated with those of Oct4 in HCC samples. Conclusions p28 GANK activates and maintains liver T-ICs in HCCs by preventing degradation of Oct4. Inhibitors of p28 GANK might therefore be developed to inactivate T-ICs and slow tumor progression.

The overall survival of patients with hepatocellular carcinoma (HCC) remains poor, and the molecular mechanisms underlying HCC progression and aggressiveness are unclear. Here, we report that increased expression of p28GANK (Gankyrin, PSMD10, or p28) in human HCC predicts poor survival and disease recurrence after surgery. Patients with HCC who have large tumors, with vascular invasion and intrahepatic or distant metastasis, expressed high levels of p28GANK. Invasive tumors overexpressing p28GANK were featured by active epithelial-mesenchymal transition (EMT), and exhibited increased angiogenesis associated with vascular endothelial growth factor overexpression, whereas silencing p28GANK expression attenuated EMT and motility/invasion of tumor cells. The p28GANK activates phosphoinositide 3-kinase (PI3K)–V-akt Murine Thymoma Viral Oncogene Homolog (AKT)–hypoxia-inducible factor 1α (HIF-1α) signaling to promote TWIST1, vascular endothelial growth factor, and metalloproteinase 2 expression. Suppression of the PI3K–AKT–HIF-1α pathway interfered with p28GANK-mediated EMT and invasion. Consistently, we detected a significant correlation between p28GANK expression and p-AKT levels in a cohort of HCC biopsies, and the combination of these two parameters is a more powerful predictor of poor prognosis. Conclusion: These results present novel mechanistic insight into a critical role of p28GANK in HCC progression and metastasis. (HEPATOLOGY 2011)

We described a combined multiple clustering approach to automatically identify chronic lymphocytic leukemia neoplastic population by flow cytometry immunophenotyping. Flow cytometry data from various specimens were preprocessed by data cross-linking and subset selection before undergoing subspace and consensus clustering. This approach was implemented as a Server-side application, with results comparable to those performed by manual gating on commercial software.

It has been shown that oncoprotein p28(GANK), which is consistently overexpressed in human hepatocellular carcinoma (HCC), plays a critical role in tumorigenesis of HCC. However, the underlying mechanism remains unclear. Here, we demonstrated that p28(GANK) inhibits apoptosis in HCC cells induced by the endoplasmic reticulum (ER) stress. During ER stress, p28(GANK) enhances the unfolded protein response, promotes ER recovery from translational repression, and thereby facilitates cell's ability to cope with the stress conditions. Furthermore, p28(GANK) upregulates glucose-regulated protein 78 (GRP78), a key ER chaperone protein, which subsequently enhances the ER folding capacity and promotes recovery from ER stress. We also demonstrated that p28(GANK) increases p38 mitogen-activated protein kinase and Akt phosphorylation, and inhibits nuclear factor kappa B (NF-kappaB) activation under ER stress, which in turn contributes to GRP78 upregulation. Taken together, our results indicate that p28(GANK) inhibits ER stress-induced apoptosis in HCC cells, at least in part, by enhancing the adaptive response and GRP78 expression. We propose that p28(GANK) has potential implications for HCC progression under the ER stress conditions.

Abstract 972 The transcriptional repressor TEL (Translocation Ets Leukemia) also known as ETV6 (Ets variant gene 6) located on human chromosome 12 is essential for hematopoietic stem cell (HSC) maintenance. Abnormalities involving TEL on the short arm of chromosome 12 including deletions and translocations are common in a surprisingly wide spectrum of hematological and non-hematological malignancies. Genome-wide analysis of leukemic samples revealed the loss of TEL expression and loss of heterozygousity (LOH) in more than half of B-cell ALL patients. We have previously reported on the generation of transgenic zebrafish expressing TEL-AML1 that develop pre-B ALL after long latency, and recapitulate the same phenotype and molecular events in human leukemias. In this study, we first used these transgenic fish to investigate the role of TEL in leukemia development. Our data shows that the loss of TEL expression contributes to, and accelerates the leukemic phenotype in our TEL-AML1 transgenic zebrafish. TEL is a selective regulator of lymphoid and HSC survival. The mechanism by which TEL modulates adult lymphoid and HSC renewal is yet to be elucidated. Moreover, studies of TEL expression and role in lymphoma development are lacking. At the genetic level, the highly conserved alternative start codon, and the initiation of translation at Methionine-43 of TEL creates a shorter isoform that is mostly nuclear. Utilizing specific antibodies that distinguish between the expression patterns of the two TEL isoforms, we demonstrate that the differential cytoplasmic and nuclear localization of TEL isoforms in human cells is involved in B- and T-cell lymphoid maturation. Our studies uncover a nuclear trafficking mechanism that is likely to regulate the nuclear repression and tumor suppressor functions of TEL. We next investigated the role of TEL in normal hematopoiesis, and TEL tumor suppressor functions in lymphoid malignancies in a novel model of zebrafish lymphoma. To study the role of TEL in hematopoiesis, we isolated the zebrafish TEL coding region and promoter elements from a BAC library, and found that TEL is expressed in early embryonic stages, in hematopoietic and neural lineages, and its expression colocalized with the hematopoietic stem cell (HSC) markers cMyb, Flk1 and RUNX1. In this model, the loss of TEL expression using morpholinos and dominant negative approaches in wild type zebrafish led to the development of lymphomas in 15% of these fish. This prompted us to investigate the expression of TEL in human lymphomas. Loss of TEL expression or the cytoplasmic localization of TEL isoforms in tumor cells from lymphoma patients were detected in eleven out of twenty three patients investigated so far (48%), and correlated with the non-Hodgkin diffuse large B-cell lymphomas and follicular subtypes as well as the aggressiveness of lymphomas. Flow cytometric analyses of lymphoma samples associated with loss of TEL expression demonstrated a CD19+ve, CD20+ve, CD10+ve, and CD45+ve phenotype. In cellular assays, we demonstrate that the activity and cellular localization of TEL are tightly regulated at multiple levels, and coordinated by P38 kinase activity. Our studies highlight a novel tumor suppressor role of TEL/ETV6 in lymphoma development, and establish TEL/ETV6 expression as a useful biomarker for non-Hodgkin lymphoma diagnosis and disease progression. Disclosures: No relevant conflicts of interest to declare.