Bromodomain extraterminal protein (BETP) inhibitors transcriptionally repress oncoproteins and nuclear factor-κB (NF-κB) target genes that undermines the growth and survival of mantle cell lymphoma (MCL) cells. However, BET bromodomain inhibitor (BETi) treatment causes accumulation of BETPs, associated with reversible binding and incomplete inhibition of BRD4 that potentially compromises the activity of BETi in MCL cells. Unlike BETi, BET-PROTACs (proteolysis-targeting chimera) ARV-825 and ARV-771 (Arvinas, Inc.) recruit and utilize an E3-ubiquitin ligase to effectively degrade BETPs in MCL cells. BET-PROTACs induce more apoptosis than BETi of MCL cells, including those resistant to ibrutinib. BET-PROTAC treatment induced more perturbations in the mRNA and protein expressions than BETi, with depletion of c-Myc, CDK4, cyclin D1 and the NF-κB transcriptional targets Bcl-xL, XIAP and BTK, while inducing the levels of HEXIM1, NOXA and CDKN1A/p21. Treatment with ARV-771, which possesses superior pharmacological properties compared with ARV-825, inhibited the in vivo growth and induced greater survival improvement than the BETi OTX015 of immune-depleted mice engrafted with MCL cells. Cotreatment of ARV-771 with ibrutinib or the BCL2 antagonist venetoclax or CDK4/6 inhibitor palbociclib synergistically induced apoptosis of MCL cells. These studies highlight promising and superior preclinical activity of BET-PROTAC than BETi, requiring further in vivo evaluation of BET-PROTAC as a therapy for ibrutinib-sensitive or -resistant MCL.
Lymphoma, Mantle-Cell , Proteins , Animals , Humans , Mice , Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Azepines/pharmacology , Cell Line, Tumor , Lymphoma, Mantle-Cell/drug therapy , Lymphoma, Mantle-Cell/metabolism , NF-kappa B/metabolism , Nuclear Proteins/metabolism , Proteins/metabolism , Proteolysis , Signal Transduction/drug effects , Thalidomide/analogs & derivatives , Thalidomide/pharmacology , Transcription Factors/metabolism
Fusion Proteins, bcr-abl/biosynthesis , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/metabolism , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/mortality , Adolescent , Adult , Aftercare , Aged , Aged, 80 and over , Disease-Free Survival , Female , Fusion Proteins, bcr-abl/genetics , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Male , Middle Aged , Survival Rate
Breast Neoplasms/therapy , Leukemia, Myeloid, Acute/etiology , Myelodysplastic Syndromes/etiology , Myelodysplastic-Myeloproliferative Diseases/etiology , Neoplasms, Second Primary/etiology , Adult , Aged , Female , Hematopoietic Stem Cell Transplantation , Humans , Leukemia, Myeloid, Acute/mortality , Leukemia, Myeloid, Acute/therapy , Middle Aged , Myelodysplastic Syndromes/mortality , Myelodysplastic Syndromes/therapy , Myelodysplastic-Myeloproliferative Diseases/mortality , Myelodysplastic-Myeloproliferative Diseases/therapy , Neoplasms, Second Primary/mortality , Neoplasms, Second Primary/therapy , Proportional Hazards Models
Leukemia, Myelogenous, Chronic, BCR-ABL Positive/drug therapy , Protein Kinase Inhibitors/therapeutic use , Protein-Tyrosine Kinases/antagonists & inhibitors , Trisomy , Adolescent , Adult , Aged , Aged, 80 and over , Child , Chromosomes, Human, Pair 8 , Female , Humans , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/genetics , Leukemia, Myelogenous, Chronic, BCR-ABL Positive/mortality , Male , Middle Aged
STAT3 is an oncogene that regulates critical cellular processes and whose constitutive activation has been demonstrated to correlate with biological and clinical features in many types of human malignancy. In this study, STAT3 activation was assessed in the Ewing sarcoma family of tumours (ESFT), which is characterized by fusion of the EWS gene with one of several Ets transcription factors, most commonly EWS-FLI1. STAT3 activation was assessed by immunohistochemistry using a monoclonal antibody specific for tyrosine(705)-phosphorylated STAT3 (pSTAT3(tyr705)) and a tissue microarray containing 49 paraffin-embedded ESFT tumours with known EWS translocations. Twenty-five (51%) tumours were pSTAT3(tyr705)-positive, as defined by more than 10% tumour cell immunostaining. STAT3 activation correlated with tumour site at presentation, with pSTAT3(tyr705)-negative ESFT involving axial sites predominantly (p = 0.008). Notably, among 31 patients who presented with localized disease, high-level STAT3 activation correlated with better overall survival (p = 0.02). STAT3 activation was not directly related to EWS-FLI1 expression, since EWS-FLI1 transfection did not result in STAT3 activation. Furthermore, detailed molecular analysis indicated that STAT3 activation may be seen with EWS-FLI1 or EWS-ERG and appears to be independent of EWS-FLI1 fusion type. In conclusion, STAT3 activation is present in approximately half of ESFT and correlates with clinical features. The role of STAT3 activation in ESFT pathogenesis seems to be independent of the type of EWS/Ets translocation.
Biomarkers, Tumor/biosynthesis , Bone Neoplasms/metabolism , Gene Expression Regulation, Neoplastic , STAT3 Transcription Factor/biosynthesis , Sarcoma, Ewing/metabolism , Adolescent , Adult , Biomarkers, Tumor/genetics , Bone Neoplasms/genetics , Bone Neoplasms/pathology , Child , Child, Preschool , Female , Humans , Immunoenzyme Techniques , Male , Neoplasm Proteins/biosynthesis , Neoplasm Proteins/genetics , Oncogene Proteins, Fusion/metabolism , Prognosis , Protein Array Analysis/methods , Proto-Oncogene Protein c-fli-1/metabolism , RNA-Binding Protein EWS/genetics , Reverse Transcriptase Polymerase Chain Reaction/methods , STAT3 Transcription Factor/genetics , Sarcoma, Ewing/genetics , Sarcoma, Ewing/pathology , Survival Analysis , Translocation, Genetic , Tumor Cells, Cultured
AIMS: In some cases distinction between chromophobe renal cell carcinoma (CRCC), oncocytoma and clear cell (conventional) renal cell carcinoma (eosinophilic variant) using routine light microscopy remains problematic. The present study investigates the level of agreement in the diagnosis of CRCC, as well as the histological features most frequently used for this diagnosis by two pathologists with a special interest in renal neoplasia. The sensitivity and specificity of immunohistochemical markers in cases with overlapping histological features in the diagnosis of CRCC were also studied. Electron microscopy was performed, as a diagnostic gold standard, on all of the cases. METHODS AND RESULTS: Thirty-two renal tumours with predominantly eosinophilic cytoplasm were reviewed in a blinded fashion by two pathologists. The diagnosis and morphological features used to render each diagnosis were tabulated. Validation of the utility of keratin 7 and 20, epithelial membrane antigen (EMA), vimentin, CD10, parvalbumin, RCC antigen, antimitochondrial antibody and Hale's colloidal iron was performed by the construction of a tissue microarray (TMA) master block. Based on histological criteria alone, overall agreement on the diagnosis of these tumours was reached in 69% of the cases, while there was total disagreement in 12%. In 59% of the cases, total agreement was reached in classifying the case as a CRCC based on histology alone. Kappa statistics for interobserver variability were calculated as only slight agreement (kappa = 0.3). The histological features most frequently associated with a diagnosis of CRCC were accentuated cell borders (87%) and a combination of hyperchromatic wrinkled nuclei (79%) and perinuclear halos (74%). The most sensitive and specific marker for CRCC was parvalbumin (sensitivity 0.91; specificity 1.0). The immunohistochemical profile of EMA+/ vimentin- was useful but had low specificity (sensitivity 0.75; specificity 0.4). CD10 had the highest sensitivity (1.0) but worst specificity (0.25) for CRCC. Keratin 7 had high sensitivity (0.83) but fairly low specificity (0.37) for CRCC. Hale's colloidal iron and the RCC antigen marker were not contributory. Finally, the antimitochondrial antibody was found to be fairly sensitive (0.83) for excluding CRCC. CONCLUSIONS: A small but significant proportion of renal tumours with cells having eosinophilic cytoplasm cannot be classified, even by experienced pathologists, based on histology alone. In these cases it is imperative to use markers with known sensitivity and specificity for the diagnosis of CRCC.
Carcinoma, Renal Cell/diagnosis , Kidney Neoplasms/diagnosis , Adenoma, Oxyphilic/diagnosis , Adenoma, Oxyphilic/metabolism , Adenoma, Oxyphilic/ultrastructure , Carcinoma, Renal Cell/metabolism , Carcinoma, Renal Cell/ultrastructure , Diagnosis, Differential , Humans , Immunohistochemistry , Intermediate Filament Proteins/analysis , Keratin-20 , Keratin-7 , Keratins/analysis , Kidney Neoplasms/metabolism , Kidney Neoplasms/ultrastructure , Microscopy, Electron , Mucin-1/analysis , Neprilysin/analysis , Observer Variation , Parvalbumins/analysis , Pathology, Clinical/standards , Prospective Studies , Retrospective Studies , Sensitivity and Specificity , Single-Blind Method , Tissue Array Analysis/methods , Vimentin/analysis
AIMS: To present three cases of sclerosing haemangioma of the lung (SHL) with prominent cystic changes. METHODS AND RESULTS: The patients were three women, 27, 35, and 64 years of age. In two cases, the lesion was found on routine chest X-ray and in both instances the diagnosis of SHL was not entertained in the radiological differential diagnosis. In one case, the finding was discovered on post mortem examination. Grossly, the lesions were well-circumscribed and cystic. Histologically, the tumours were characterized by a dual population of small and large bland-appearing neoplastic cells growing in a predominantly cystic pattern. Immunohistochemical staining for epithelial membrane antigen by tumour cells was consistently positive in all three cases. All tumours were negative for keratin, CD34, factor VIII, and S100. The two patients in whom the lesion was identified ante-mortem are alive and free of disease 4 and 7 years after surgical excision. CONCLUSIONS: The present cases indicate that sclerosing haemangioma may present as a cystic pulmonary neoplasm; such a presentation should be taken into consideration when assessing cystic pulmonary lesions.
Cysts/pathology , Hemangioma/pathology , Lung Neoplasms/pathology , Adult , Aged , Diagnosis, Differential , Female , Hemangioma/metabolism , Humans , Immunohistochemistry , Lung Neoplasms/metabolism , Mucin-1/metabolism , Sclerosis/pathology
