Expression of the cerbB-2 (HER-2/neu) oncoprotein in prostatic adenocarcinoma

We report a 62 years old kidney transplant (KT) patient who was diagnosed of localized prostatic cancer (PC) after 6 years of the implant. Transrectal prostatic High Intensity Focused Ultrasound (HIFU) was applied. Results have been satisfactory, achieving pathologic and biochemical success. The discharge was completed at 24 hs, the morbidity was minimal. We have not found any reference in the literature on the appliance of HIFU in PC KT patients. We think that HIFU may represent a good alternative for these patients.     

Expression of NEU/HER-2 oncoprotein (p185neu) in prostate tumors: An immunohistochemical study

The expression of the neu oncogene product was investigated immunohistochemically in 36 cases of benign prostatic hyperplasia (BPH) and seven cases of adenocarcinoma of prostate (CaP). c-neu oncogene encodes a transmembrane growth factor receptor that has partial structural homology with EGF receptor, and is overexpressed and amplified in a number of human tumors, specially, breast cancers. Using a monoclonal antibody, AB-3, which recognizes -COOH-terminal of neu oncoprotein, we have analyzed immunohistochemically the expression of this protein in buffered formalin and Zamboni fluid-fixed surgically removed tissues. Focal patchy and/or diffused cytoplasmic staining of varying intensity was observed in 34 of 36 BPH cases. Four cases showed cell membrane staining as well (4/36 = 11%). All seven cases of adenocarcinomas had moderate to strong c-neu immunoreactivity, and two gave a distinct cell membrane-positive reaction (100%). The available data indicate that prostatic tumors as well as a high percentage of prostatic hyperplasia tissues express c-neu protein; however, its role in cellular proliferation needs further study. © 1993 Wiley-Liss, Inc.     

Urokinase Receptor Mediates Osteoclastogenesis via M‐CSF Release From Osteoblasts and the c‐Fms/PI3K/Akt/NF‐κB Pathway in Osteoclasts

Bone remodeling is a dynamic process based on a fine‐tuned balance between formation and degradation of bone. Osteoblasts (OBLs) are responsible for bone formation and bone resorption is mediated by osteoclasts (OCLs). The mechanisms regulating the OBL‐OCL balance are critical in health and disease; however, they are still far from being understood. We reported recently that the multifunctional urokinase receptor (uPAR) mediates osteogenic differentiation of mesenchymal stem cells (MSCs) to OBLs and vascular calcification in atherosclerosis. Here, we address the question of whether uPAR may also be engaged in regulation of osteoclastogenesis. We show that uPAR mediates this process in a dual fashion. Thus, uPAR affected OBL‐OCL interplay. We observed that osteoclastogenesis was significantly impaired in co‐culture of monocyte‐derived OCLs and in OBLs derived from MSCs lacking uPAR. We show that expression and release, from OBLs, of macrophage colony‐stimulating factor (M‐CSF), which is indispensable for OCL differentiation, was inhibited by uPAR loss. We further found that uPAR, on the other hand, controlled formation, differentiation, and functional properties of macrophage‐derived OCLs. Expression of osteoclastogenic markers, such as tartrate‐resistant acid phosphatase (TRAP) and cathepsin K, was impaired in OCLs derived from uPAR‐deficient macrophages. The requirement of uPAR for osteoclastogenesis was further confirmed by immunocytochemistry and in bone resorption assay. We provide evidence that the underlying signaling mechanisms involve uPAR association with the M‐CSF binding receptor c‐Fms followed by c‐Fms phosphorylation and activation of the PI3K/Akt/NF‐κB pathway in OCLs. We further show that uPAR uses this pathway to regulate a balance between OCL differentiation, apoptosis, and cell proliferation. Our study identified uPAR as an important and multifaceted regulator of OBL‐OCL molecular interplay that may serve as an attractive target in bone disease and ectopic calcification. © 2014 American Society for Bone and Mineral Research.     

The efficacy and safety of hypofractionated radiotherapy with concurrent anti‐HER‐2 therapy following breast‐conserving therapy for breast cancer

The purpose of this study was to report rates and severities of radiation‐related toxicities and analyze disease‐control outcomes in patients who have received hypofractionated whole breast radiation (HF) with concurrent trastuzumab with or without pertuzumab. We conducted a retrospective cohort study including women with stage I‐III HER2‐positive breast cancer who received HF at the University of Pennsylvania between 1/2005 and 5/2018 with concurrent trastuzumab with or without pertuzumab. Fractionation was 266 cGy daily to a total dose of 4256 cGy with or without a sequential tumor bed boost. Eighty patients were included in the cohort with a median follow‐up time of 21.44 months. There was one grade 3 acute toxicity (fatigue) and no grade 3 late toxicities. 91% and 25% of patients experienced grade 1‐2 acute and late skin reactions, respectively. An excellent‐good cosmetic outcome was reported by 74% and 95% of patients and physicians, respectively. No patients experienced tumor recurrences, and the only death was due to a secondary cause. These results suggest that hypofractionated whole breast radiation administered concurrently with anti‐HER‐2 therapies is efficacious and has acceptable toxicity in early‐stage breast cancer patients treated with lumpectomy. Continued follow‐up is warranted to evaluate long‐term outcomes.     

C-erb-B2 (HER2/neu) expression in synovial sarcoma of the head and neck

BACKGROUND: Synovial sarcoma is a malignant mesenchymal tumor composed of varying proportions of spindle and epithelial cell components. Because of the histologic and immunohistochemical similarity of synovial sarcoma to epithelial carcinomas, we hypothesized that the human epithelial growth factor receptor 2 (C-erb-B2, also termed HER2/neu) may contribute to the tumor phenotype and provide a new therapeutic target for this soft tissue tumor. METHODS: Three head and neck, one chest wall, and seven extremity synovial sarcomas were evaluated for C-erb-B2 (HER2/neu) expression by immunohistochemistry, Western immunoblotting, and fluorescence in situ hybridization (FISH). RESULTS: The head and neck cases demonstrated immunohistochemically strong positive staining, whereas tumors from other anatomic locations showed neither positive nor cytoplasmic restricted staining. Antigen-targeted antibody therapy (trastuzumab) was initiated in two patients. CONCLUSIONS: These results demonstrate that C-erb-B2 (HER2/neu) may play a role in the tumorigenesis of synovial sarcoma; and, therefore, antigrowth factor therapies may provide a previously unrecognized pharmaceutical approach to soft tissue tumors. The data also suggest that although synovial sarcoma of the head and neck and synovial sarcoma of the extremities have similar morphologic features, they may be clinically and mechanistically distinct entities.     

Retrospective Study of 156 Atrial Septal Defects in Dogs and Cats (2001–2005)

Atrial septal defect (ASD) is a common congenital heart disease (CHD) in humans, but considered relatively rare in veterinary medicine. However, modern echocardiographic and Doppler techniques currently offer a good view of the morphology of the interatrial septum, thus facilitating earlier detection of ASD in awake animals. In this context, we carried out a retrospective study of cases of recently diagnosed ASD in dogs and cats at the Cardiology Unit of Alfort (2001–2005) using echocardiography combined with colour Doppler mode. ASD was diagnosed in 156 animals and represented 37.7% of all canine and feline CHDs (n = 414). ASD was the most common CHD after mitral dysplasia in both species. Boxer and Domestic shorthair were the most common canine and feline breeds affected. Most defects (98.7%) were secundum‐type ASD, without clinical signs in 73.7% of cases. The most common clinical signs included systolic murmur heard over the left heart base (20.2%), exercise intolerance (7.0%), syncope (5.3%), dyspnoea (2.6%) and cough (2.6%). Animals that presented a systolic heart murmur over the left base had a significantly larger ASD than others (P < 0.05). These data suggest that the incidence of ASD is higher than previously assumed. ASD should be suspected, for example, in instances of left basal systolic heart murmur, although its clinical and haemodynamic consequences are usually minor.     

CCN family 2/connective tissue growth factor (CCN2/CTGF) promotes osteoclastogenesis via induction of and interaction with dendritic cell–specific transmembrane protein (DC‐STAMP)

CCN family 2/connective tissue growth factor (CCN2/CTGF) promotes endochondral ossification. However, the role of CCN2 in the replacement of hypertrophic cartilage with bone is still unclear. The phenotype of Ccn2 null mice, having an expanded hypertrophic zone, indicates that the resorption of the cartilage extracellular matrix is impaired therein. Therefore, we analyzed the role of CCN2 in osteoclastogenesis because cartilage extracellular matrix is resorbed mainly by osteoclasts during endochondral ossification. Expression of the Ccn2 gene was upregulated in mouse macrophage cell line RAW264.7 on day 6 after treatment of glutathione S transferase (GST) fusion mouse receptor activator of NF‐κB ligand (GST‐RANKL), and a combination of recombinant CCN2 (rCCN2) and GST‐RANKL significantly enhanced tartrate‐resistant acid phosphatase (TRACP)–positive multinucleated cell formation compared with GST‐RANKL alone. Therefore, we suspected the involvement of CCN2 in cell‐cell fusion during osteoclastogenesis. To clarify the mechanism, we performed real‐time PCR analysis of gene expression, coimmunoprecipitation analysis, and solid‐phase binding assay of CCN2 and dendritic cell–specific transmembrane protein (DC‐STAMP), which is involved in cell‐cell fusion. The results showed that CCN2 induced and interacted with DC‐STAMP. Furthermore, GST‐RANKL–induced osteoclastogenesis was impaired in fetal liver cells from Ccn2 null mice, and the impaired osteoclast formation was rescued by the addition of exogenous rCCN2 or the forced expression of DC‐STAMP by a retroviral vector. These results suggest that CCN2 expressed during osteoclastogenesis promotes osteoclast formation via induction of and interaction with DC‐STAMP. © 2011 American Society for Bone and Mineral Research.     

Disengaging the IL‐2 Receptor with Daclizumab Enhances IL‐7–Mediated Proliferation of CD4+ and CD8+ T Cells

Allograft rejection is mainly driven by the production of IL‐2, which expands T cells by linking the IL‐2 receptor (IL‐2R) composed of three subunits: CD25, CD122 and CD132. Daclizumab, widely used in immunosuppression, is a humanized anti‐CD25 antibody that disrupts IL‐2 signaling by binding to CD25 and preventing the assembly of the high‐affinity IL‐2R. Here we show that Daclizumab, while blocking the T‐cell response to IL‐2, increases CD4⁺ and CD8⁺ T‐cell proliferative response to the homeostatic cytokine IL‐7. The IL‐7R shares CD132 with the IL‐2R and blocking of CD25 by Daclizumab results in the enhanced formation of the IL‐7R that in turn allows IL‐7 to bind more efficiently on the cell surface. The consequently increased IL‐7R signaling boosts intracellular phosphorylated STAT5 and T‐cell proliferation. In addition, treatment with Daclizumab delays the internalization of CD127 upon IL‐7 treatment, retaining T‐cell sensitivity to IL‐7 for a prolonged time. This effect of Daclizumab highlights the redundancy of the cytokine system, which may influence T‐cell proliferation in transplanted patients, and provides information to improve future immunosuppressive strategies.     

Repair of large osteochondral defects in rabbits using porous hydroxyapatite/collagen (HAp/Col) and fibroblast growth factor‐2 (FGF‐2)

Articular cartilage has a limited capacity for self‐renewal. This article reports the development of a porous hydroxyapatite/collagen (HAp/Col) scaffold as a bone void filler and a vehicle for drug administration. The scaffold consists of HAp nanocrystals and type I atelocollagen. The purpose of this study was to investigate the efficacy of porous HAp/Col impregnated with FGF‐2 to repair large osteochondral defects in a rabbit model. Ninety‐six cylindrical osteochondral defects 5 mm in diameter and 5 mm in depth were created in the femoral trochlear groove of the right knee. Animals were assigned to one of four treatment groups: porous HAp/Col impregnated with 50 µl of FGF‐2 at a concentration of 10 or 100 µg/ml (FGF10 or FGF100 group); porous HAp/Col with 50 µl of PBS (HAp/Col group); and no implantation (defect group). The defect areas were examined grossly and histologically. Subchondral bone regeneration was quantified 3, 6, 12, and 24 weeks after surgery. Abundant bone formation was observed in the HAp/Col implanted groups as compared to the defect group. The FGF10 group displayed not only the most abundant bone regeneration but also the most satisfactory cartilage regeneration, with cartilage presenting a hyaline‐like appearance. These findings suggest that porous HAp/Col with FGF‐2 augments the cartilage repair process. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 28:677–686, 2010     

Glycoprotein CD44 Expression in Human Breast Cancer: An Immunohistochemical Study Including Correlation with Cathepsin D, Type IV Collagen, Laminin, Fibronectin, EGFR, c-erbB-2 Oncoprotein, p53, Steroid Receptor Status, and Proliferative Indices

Abstract: CD44, an integral membrane glycoprotein has diverse functions in cell-cell and cell-matrix interactions and may be a determinant of metastatic and invasive behavior in carcinomas. The expression of CD44 in 86 female breast carcinomas was examined by immunohistochemistry using the monoclonal mouse antihuman phagocytic glycoprotein-1, CD44 (clone DF 1485) on formalin-fixed, paraffin-embedded tissue. The CD44 expression was studied in correlation with the expression of basement membrane (BM) antigen (type IV collagen, laminin), fibronectin, cathepsin D, p53, c-erbB-2, proliferative activity Ki-67, PCNA), steroid receptor content, as well as with other conventional clinicopathological parameters in breast cancer. CD44 expression was observed in variable amounts in the epithelial cells of the control group (cases of fibrocystic and benign proliferative-breast disease). CD44 expression showed a heterogeneous pattern, such as small foci amounting to less than 5% of the total cells in a given section, or larger clusters of tumor cells weakly or strongly stained. A greater proportion of the tumor stained in a patchy pattern. Seventy-two point six percent of tumors showed CD44 expression (>5% of neoplastic cells). In 10.9% of the cancer, more than 90% of the cancer cells stained positively for CD44. High values of CD44 were correlated with lymphnode involvement (p = 0.006) and progesterone receptor content (p = 0.02). There was no significant correlation between CD44 expression and the other clinicopathological parameters, including tumor size, age of patient, histological type, tumor grade and estrogen receptor content as well as with the other prognostic parameters.
These findings suggest that the CD44 expression in breast cancer is independent of the expression of extracellular components, the proteolytic enzyme cathepsin D, and the growth fraction of tumor, but may be useful for monitoring human-breast metastasis.     

Assessment of Tocilizumab (Anti–Interleukin‐6 Receptor Monoclonal) as a Potential Treatment for Chronic Antibody‐Mediated Rejection and Transplant Glomerulopathy in HLA‐Sensitized Renal Allograft Recipients

Extending the functional integrity of renal allografts is the primary goal of transplant medicine. The development of donor‐specific antibodies (DSAs) posttransplantation leads to chronic active antibody‐mediated rejection (cAMR) and transplant glomerulopathy (TG), resulting in the majority of graft losses that occur in the United States. This reduces the quality and length of life for patients and increases cost. There are no approved treatments for cAMR. Evidence suggests the proinflammatory cytokine interleukin 6 (IL‐6) may play an important role in DSA generation and cAMR. We identified 36 renal transplant patients with cAMR plus DSAs and TG who failed standard of care treatment with IVIg plus rituximab with or without plasma exchange. Patients were offered rescue therapy with the anti–IL‐6 receptor monoclonal tocilizumab with monthly infusions and monitored for DSAs and long‐term outcomes. Tocilizumab‐treated patients demonstrated graft survival and patient survival rates of 80% and 91% at 6 years, respectively. Significant reductions in DSAs and stabilization of renal function were seen at 2 years. No significant adverse events or severe adverse events were seen. Tocilizumab provides good long‐term outcomes for patients with cAMR and TG, especially compared with historical published treatments. Inhibition of the IL‐6–IL‐6 receptor pathway may represent a novel approach to stabilize allograft function and extend patient lives.