Worse prognosis of osteosarcoma patients expressing IGF-1 on a tissue microarray

BACKGROUND It is hardly possible to define osteosarcoma (OS) patients at greatest risk for non-response to chemotherapy, metastasis and short survival times. Our goal was the investigation of local expression of insulin-like growth factor (IGF-1) with regard to survival time of OS patients using a tissue microarray (TMA). MATERIALS AND METHODS Tumor tissue specimens from surgical primary tumor resections were collected from patients with OS. A TMA was composed, sections were stained with rabbit anti-IGF-1 and grading was performed. Statistics involved Kaplan-Meier curves and the log-rank test. RESULTS We analyzed immunohistochemical expression of local IGF-1 on a TMA based on surgical primary tumor resections of 67 OS patients. The mean clinical follow-up time was 98 months. Twenty-two (33%) OS patients stained negatively and 44 (66%) OS patients stained positively for IGF-1. Significantly shorter survival was detected with expression of IGF-1 (p=0.007). The 5-year survival rate for patients expressing IGF-1 was 63% compared to 92% in patients without expression of IGF-1. Non-responders to chemotherapy and patients with metastasis, who also stained positively for IGF-1 manifested a significantly (p=0.002 and p<0.0001, respectively) shorter survival. CONCLUSION Expression of local IGF-1 in primary tumor tissue appears to significantly affect the aggressiveness of OS, may predict survival time and, above all, may discriminate patients with non-response to chemotherapy and metastasis. This represents the basis for successful patient selection with regard to the decision process for or against chemotherapy and the choice of the most effective therapeutic drug. It may be a more important marker of tumor progression and indicator of prognosis than serum IGF-1. Novel tumor markers and therapeutic agents targeting the local IGF-1 pathway may increase the likelihood of therapeutic success

Anwendung von Allgemeintoleranzen im ISO GPSNormensystem – Aspekte der Normung

Bei der Erstellung der Produktdokumentation nach dem ISO GPS-Normensystem sind die Normen ISO 22081 und DIN 2769 bezüglich der Angabe von Allgemeintoleranzen anzuwenden. Beide Normen erreichen in der Anwendung noch nicht die Akzeptanz der Vorläufernormen ISO 2768, Teile 1 und 2. Dies gilt insbesondere, wenn auf die Erstellung einer Zeichnung verzichtet wird und die geometrische Spezifikation ausschließlich im 3DCAD-Modell und dem zugeordneten Datensatz erstellt wird. Gründe sind neben ungenügendem Wissen in der Anwendung der GPS-Normen auch Limitierungen der in den Normen angegebenen Regeln für die Anwendbarkeit von Allgemeintoleranzen. Darüber hinaus bestehen begriffliche Unklarheiten innerhalb des GPS-Normensystems sowie zwischen dem GPS-Normensystem und den Normen zur Technischen Produktdokumentation. Im Interesse einer aufwandsarmen Erstellung der Produktdefinition ist die weitreichende Anwendung von Allgemeintoleranzen erstrebenswert, da sie die Anzahl der erforderlichen individuellen Toleranzspezifikationen teils deutlich reduziert. Der Beitrag identifiziert einige normative Festlegungen, die das einschränken bzw. dem Entwickler derzeit wenig Hilfe bei der Festlegung der Allgemeintoleranzen geben. Ansätze für die Verbesserung werden vorgeschlagen

Foscan and Foslip based photodynamic therapy in osteosarcoma in vitro and in intratibial mouse models

Current osteosarcoma therapies cause severe treatment-related side effects and chemoresistance, and have low success rates. Consequently, alternative treatment options are urgently needed. Photodynamic therapy (PDT) is a minimally invasive, local therapy with proven clinical efficacy for a variety of tumor types. PDT is cytotoxic, provokes anti-vascular effects and stimulates tumor cell targeting mechanisms of the immune system and, consequently, has potential as a novel therapy for osteosarcoma patients. This study investigated the uptake and the dark- and phototoxicity and cytotoxic mechanisms of the photosensitizer (PS) 5,10,15,20-tetrakis(meta-hydroxyphenyl) chlorine (mTHPC, Foscan) and a liposomal mTHPC formulation (Foslip) in the human 143B and a mouse K7M2-derived osteosaroma cell line (K7M2L2) in vitro. Secondly the tumor- and metastasis-suppressive efficacies of mTHPC formulations based PDT and associated mechanisms in intratibial, metastasizing osteosarcoma mouse models (143B/SCID and syngeneic K7M2L2/BALB/c) were studied. The uptake of Foscan and Foslip in vitro was time- and dose-dependent and resulted in mTHPC and light dose-dependent phototoxicity associated with apoptosis. In vivo, the uptake of both i.v. administered mTHPC formulations was higher in tumor than in healthy control tissue. PDT caused significant (Foscan P<0.05, Foslip P<0.001) tumor growth inhibition in both models. A significant (Foscan P<0.001, Foslip P<0.001) immunsystem-dependent suppression of lung metastasis was only observed in the K7M2L2/BALB/c model and was associated with a marked infiltration of T-lymphocytes at the primary tumor site. This article is protected by copyright. All rights reserved

Linking thermal and seismic mantle structure in the light of uncertain mineralogy and limited tomographic resolution

&amp;lt;p&amp;gt;Mantle convection is primarily driven by gravitational forces acting on thermally buoyant structures in Earth&amp;#039;s interior. The associated vertical stresses generate phases of uplift and subsidence of the surface, leaving observable traces in the geologic record. Utilizing new data assimilation techniques, geodynamic inverse models of mantle flow can provide theoretical estimates of these surface processes, which can be tested against geologic observations. These so-called mantle flow retrodictions are emerging as powerful tools that have the potential to allow for tighter constraints on the inherent physical parameters.&amp;lt;/p&amp;gt; &amp;lt;p&amp;gt;To contain meaningful information, the inverse models require an estimate of the present-day buoyancy distribution within the mantle, which can be derived from seismic observations. By using thermodynamically self-consistent models of mantle mineralogy, it is possible to convert the seismic structure of global tomographic models to temperature. However, both seismic and mineralogical models are significantly affected by different sources of uncertainty and often require subjective modelling choices, which can lead to different estimated properties. In addition, due to the complexity of the mineralogical models, the relation between temperature and seismic velocities is highly nonlinear and not strictly bijective: In the presence of phase transitions, different temperatures can result in the same seismic velocity, further complicating the conversion between the two parameters.&amp;lt;/p&amp;gt; &amp;lt;p&amp;gt;&amp;amp;#160;&amp;lt;/p&amp;gt; &amp;lt;p&amp;gt;Using a synthetic closed-loop experiment, we investigate the theoretical ability to estimate the present-day thermal state of Earth&amp;#039;s mantle based on tomographic models. The temperature distribution from a 3-D mantle circulation model with earth-like convective vigour serves as a representation of the &amp;quot;true&amp;quot; temperature field, which we aim to recover after a set of processing steps. These steps include the &amp;amp;#8220;forward and inverse&amp;amp;#8221; mineralogical mapping between temperatures and seismic velocities, using a thermodynamic model for pyrolite composition, as well as applying a tomographic filter to mimic the limited resolution and uneven data coverage of the underlying tomographic model. Owing to imperfect knowledge of the parameters governing mineral anelasticity, we test the effects of changes to the anelastic correction applied in forward and inverse mineralogical mapping. The mismatch between the recovered and the initial temperature field carries a strong imprint of the tomographic filter. Additionally, we observe systematic errors in the recovered temperature field in the vicinity of phase transitions. Our results highlight that, given the current limits of tomographic models and the incomplete knowledge of mantle mineralogy, amplitudes and spatial scales of a temperature field obtained through global seismic models will deviate significantly from the true state. Strategies to recover the present-day buoyancy field must be carefully selected in order to minimize additional uncertainties.&amp;lt;/p&amp;gt;</jats:p

Expression of MSH2 and MSH6 on a Tissue Microarray in Patients with Osteosarcoma

BACKGROUND/AIM: Reliable prognostic factors for the outcome of patients with osteosarcoma (OS) remain elusive. We analyzed the relationship between immunohistochemical expression of deoxyribonucleic acid (DNA) mismatch repair (MMR) proteins, MutS protein homolog 2 (MSH2) and MSH6 using a tissue microarray (TMA) with respect to OS patient demographics and survival time. MATERIALS AND METHODS: We retrieved tumor tissue specimens from bone tissue originating from surgical primary tumor specimens of OS patients to generate a TMA and stained sections with antibodies against MSH2 and MSH6. RESULTS: Tumor resections of 67 patients with a mean follow-up of 98 months were evaluated. MSH2 was expressed in nine (13%), MSH6 in ten (15%) and combined MSH2 and MSH6 (MSH2/6) in six (9%) patients. Significantly shorter survival times were associated with expression of MSH6, MSH2/6, as well as simultaneous non-response to chemotherapy and presence of metastasis. CONCLUSION: The survival time of patients with OS may be predicted by local expression of MSH6 and MSH2/6 in surgical primary tumor resections. This study shows the prognostic value of the local expression of DNA MMR proteins, as markers for poor prognosis of OS patients

Abstract 589: Targeting of metastatic osteosarcoma with F8-TNF-alpha in an orthotopic mouse model

Abstract Osteosarcoma (OS) is a malignancy of bone and five-year survival rates are still at a low 20% if metastatic disease is present. Metastatic OSs most frequently metastasize to the lung. TNF-α is a powerful cytokine already shown to be active against many cancers. Nevertheless, TNF-α is highly toxic at clinically relevant concentrations. To overcome this limitation, targeted forms of TNF-α can be used. In this study, targeting is achieved through linking TNF-α to the antibody fragment F8, targeting the EDA-domain of fibronectin. To test the efficacy of F8-TNF-α against OS, we sought to investigate the effects of F8-TNF-α in an orthotopic syngeneic OS model. In addition to evaluating compound efficacy, we also tested whether the route of administration (systemic versus local application) can even further increase treatment efficacy. Ultimately, we aimed at measuring treatment success by assessing different stages of OS progression, especially metastatic disease. To show clinical relevance, we performed immunofluorescence (IF) of EDA on primary human OS samples. In a second step, we evaluated drug efficacy in a syngeneic OS mouse model. K7M2L2 OS cells, stably infected to express the marker proteins lacZ and mCherry, were orthotopically injected into the tibia of mice. Once primary tumors were detectable, the mice were treated with either PBS or F8-TNF-α. The compound was given either systemically (intravenously (i.v.), tail vein) or locally (intraarterial (i.a.), femoral artery). Four days after the last treatment, all mice were sacrificed, terminal blood sampling was performed and blood samples were analyzed for the presence of circulating tumor cells (CTCs) via FACS/mCherry fluorescence. Following X-gal staining, metastases on the surface of lungs were counted. EDA and infiltration of immune cells (CD4+, CD8+, natural killer (NK) cells, CD19+) were assessed histologically. Using IF, we detected strong expression of EDA in primary tumors as well as bone and lung metastases from human OS patients. Preliminary results from our treatment study showed a significant decrease in the number of lung metastases as well as the number of CTCs in mice treated with i.a. F8-TNF-α while the treatment was well tolerated by the mice. The reduction of lung metastases could, at least in part, be explained by a strong expression of EDA in the lung metastases. In addition, significant increases in lung-infiltrating immune cells were observed after administration of F8-TNF-α (i.v. F8- TNF-α: increased CD4+ T-cells; i.a. F8-TNF-α: increased NK cells). Although significant reductions in numbers of metastases and CTCs were detected, F8-TNF-α did not significantly impair the growth of the primary tumors. In conclusion, our study demonstrates a reduction of CTCs as well as lung metastases after administration of F8-TNF-α and thus, provides a rationale for further studying the anti-metastatic effects of this compound in a clinically relevant model of OS. Citation Format: Bernhard Robl, Sander Martijn Botter, Aleksandar Boro, Dario Neri, Bruno Fuchs. Targeting of metastatic osteosarcoma with F8-TNF-alpha in an orthotopic mouse model. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 589.</jats:p