Prognostic Factor In Osteosarcoma Research Articles

Abstract 342: Telomerase-dependent oncolytic adenovirus sensitizes human osteosarcoma cells to chemotherapy through Mcl-1 downregulation

Abstract Osteosarcoma is the most common malignant primary bone tumor. Despite advances in multi-agent chemotherapy, poor response to chemotherapy is one of the critical prognostic factors in osteosarcoma. Therefore, enhancement of chemosensitivity is one approach to improve the survival of osteosarcoma patients. We recently developed a telomerase-specific replication-competent oncolytic adenovirus, Telomelysin (OBP-301). A Phase I clinical trial in the U.S. showed the safety of OBP-301 in advanced cancer patients, and a Phase I/II clinical trial for malignant thoracic tumors is currently undergoing. We recently confirmed the antitumor effect of OBP-301 monotherapy in human osteosarcoma cells. In this study, we investigated the chemosensitizing effect of OBP-301 in human osteosarcoma cells and the molecular mechanism in the OBP-301-mediated enhancement of cell death. We used four human osteosarcoma cell lines, HOS, MNNG/HOS, 143B and SaOS-2. OBP-301 is an attenuated adenovirus, in which the hTERT promoter drives the expression of E1 gene, and causes tumor-selective lysis in a variety of human malignant tumor cells with telomerase activity. OBP-301 infection enhanced the cytotoxic effect of chemotherapeutic agents, cisplatin and doxorubicin, that are commonly used for the treatment of osteosarcomas. The calculation of combination index revealed the synergistic effects in all human osteosarcoma cell lines. Combination of OBP-301 increased apoptosis in the chemotherapeutic agents-treated cells. To clarify the molecular mechanism for the chemosensitizing effect of OBP-301, the expression of Bcl-2 family proteins, which are critical regulators of apoptosis, were investigated. In OBP-301-infected MNNG/HOS and SaOS-2 cells, the expression level of Mcl-1 was markedly decreased by OBP-301 infection. Downregulation of Mcl-1 expression by siRNA enhanced the chemotherapeutic agents-induced apoptosis. Moreover, combination therapy of chemotherapeutic agents with OBP-301 significantly suppressed tumor growth in a subcutaneous xenograft tumor model compared to monotherapy with chemotherapeutic agents or OBP-301. Anti-apoptotic Mcl-1 protein has been shown to be frequently overexpressed in human sarcomas. Overexpression of anti-apoptotic proteins is an important factor to prevent chemotherapy-induced apoptosis in cancer. We demonstrated the involvement of Mcl-1 suppression in the chemosensitizing effect of OBP-301 in human osteosarcoma cells. These results suggest that Mcl-1 is a critical target for improvement of the chemosensitivity in human osteosarcoma cells and the combination of chemotherapy with OBP-301 may be a novel therapeutic strategy for osteosarcoma patients. Citation Format: Shuhei Osaki, Toshinori Omori, Hiroshi Tazawa, Joe Hasei, Yasuaki Yamakawa, Tsuyoshi Sasaki, Toshiyuki Kunisada, Yasuo Urata, Toshifumi Ozaki, Toshiyoshi Fujiwara. Telomerase-dependent oncolytic adenovirus sensitizes human osteosarcoma cells to chemotherapy through Mcl-1 downregulation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 342. doi:10.1158/1538-7445.AM2014-342

Abstract 3102: Genomic alterations in biopsy samples predict the chemosensitivity of pediatric osteosarcoma

Abstract PURPOSE: Sensitivity of preoperative chemotherapy, evaluated by a histological analysis using resected tumor tissue, is the most important prognostic factor for osteosarcoma (OS). Methotrexate, doxorubicin, cisplatin (MAP regimen), and ifosfamide (IFO) are recognized as key drugs for OS, although it is still debatable whether addition of IFO for all OS patient is beneficial or not. In this study, to construct a prediction system of chemosensitivity of preoperative chemotherapy for pediatric OS patients, array CGH and NGS technologies were applied. METHODS: Pre-therapeutic biopsy tumor samples of 30 pediatric conventional OS patients treated in our institute who had homogeneous clinicopathological- and therapeutic- background were analyzed. The patients were divided into three groups (A:MAP-sensitive, B:IFO-sensitive and C:resistant to both) according to the histological response to chemotherapy including MAP and/or IFO. Twenty-two tumor samples were used as a learning set for searching candidate markers. Additionally, 8 tumor samples were used for independent analysis as a validation set. Genomic DNAs were prepared from frozen biopsy samples and used for CGH analysis with Agilent 44k DNA microarray. Target-sequencing by using the Ion Torrent AmpliSeq Comprehensive Cancer Panel containing 409 cancer-related genes were also conducted for 10 tumor/non-tumor pairs. RESULTS: Survival analysis of this cohort assured that response levels to chemotherapy were significantly correlated to the patient outcomes. Differential aberration analyses of the copy number data resulted in selection of 8 putative genomic markers for classifying into A, B and C groups with differential chemosensitivity. Scoring system to predict chemosensitivity of each tumor was developed with these genomic markers and subsequent validation analysis indicated that the scoring system was remarkably consistent to the patient properties. Target re-sequencing of 409 cancer-related genes by using Ion Proton sequencer identified 98 candidate non-synonymous single nucleotide variations (SNVs) (average 10 SNVs per tumor), however, significantly recurrent mutations in C group have not been observed so far. CONCLUSION: Our results indicated that copy number changes in OS biopsy samples could be good predictors of response to preoperative chemotherapy. These signatures as well as global mutation information may be useful for the future genomics-based personalized chemotherapy for OS patients. Citation Format: Shintaro Iwata, Tsukasa Yonemoto, Hajime Kageyama, Hiroto Kamoda, Sana Yokoi, Hiroki Nagase, Akira Nakagawara, Takeshi Ishii, Miki Ohira. Genomic alterations in biopsy samples predict the chemosensitivity of pediatric osteosarcoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 3102. doi:10.1158/1538-7445.AM2014-3102

Postoperative nomogram to predict the probability of metastasis in Enneking stage IIB extremity osteosarcoma.

BackgroundMetastasis is the most crucial prognostic factor in osteosarcoma. The goal of this study was to develop a new nomogram to predict the probability of metastasis in Enneking stage IIB extremity osteosarcoma after neoadjuvant chemotherapy and limb salvage surgery.MethodsWe examined medical records of 91 patients who had undergone surgery between March 1994 and March 2007. A nomogram was developed using multivariate logistic regression. The nomogram was validated internally by bootstrapping-method (200 repetitions) and externally in independent validation set (n = 34). A Youden-derived cutoff value was assigned to the nomogram to predict dichotomous outcomes for metastasis.ResultsThe nomogram was built from four predictors of tumor site, serum alkaline phosphatase, intracapsular extension, and Huvos grade, and an additional clause that the cutoff value should be added to the total points in the cases of incomplete surgical resection. P-value of Hosmer and Lemshow Goodness-of-fit test of this model was 0.649. Area under receiver operating curve values of 0.83 (95% confidence interval [CI], 0.75 to 0.92) in the training set and 0.80 (95% CI, 0.63 to 0.96) in the validation set were obtained. The accuracy of dichotomous outcomes was 79.1% (95% CI, 0.69 to 0.86) and 82.4% (95% CI, 0.63 to 0.92) in the training and validation sets.ConclusionsWe have developed a new high-performance nomogram to predict the probability of metastasis in Enneking stage IIB extremity osteosarcoma after limb salvage surgery.

Changing prognostic factors in osteosarcoma: analysis of 381 cases from two institutions.

Osteosarcoma occurs most commonly in children and young adults, with a historic second incidence peak in the elderly. Most studies have focused on those occurring in adolescence. Detailed information on descriptive features and prognostic factors in patients of different age groups is lacking. We analyzed 381 osteosarcomas diagnosed between 1973 and 2012 to identify factors significantly associated with survival in various age groups. The peak incidence was seen in patients age <25, followed by a steady incidence rate thereafter until the sixth decade, when it started to decline. In the early onset diseases, significant factors for recurrence-free survival (RFS) were tumor site and size; whereas those for overall survival (OS) were gender, tumor site, type, grade and size. In patients age 25 to 54, tumor type and grade were significant for RFS, and the pathologic type was significant for OS. In those age ≥55, race and tumor size were significant for RFS; tumor site and size were significant for OS. In multivariate analysis, tumor size remained significant for RFS; gender, tumor site and size maintained their significance for OS in patients age <25. While no independent factor was identified in patients age 25 to 54, tumor size remained significant for RFS in those age ≥55. Chemotherapy-induced tumor necrosis was a prognosticator for RFS in patients age 25 to 54 by univariate analysis, but not as an independent factor in any stratified age group. Our data indicate that the distinctive prognostic factors differed significantly among different age groups, thus providing a rationale for age-based management strategies.

Is it important to maintain high-dose intensity chemotherapy in the treatment of adults with osteosarcoma?

Neoadjuvant chemotherapy for osteosarcoma is the standard of care, but there is still confusion regarding the best chemotherapy regimen and the optimal intensity. This retrospective study intends to evaluate whether there is a clear correlation between the chemotherapy dose intensity (DI) and the percentage of tumor necrosis, the risk of tumor recurrence after surgery and patient survival. The medical records of all adult patients with localized osteosarcoma that received treatment between the years of 1998 and 2009 at the Tel Aviv Sourasky Medical Center were analyzed. We used multiple logistic/linear regression models to test the effect of the neoadjuvant chemotherapy relative DI (RDI) on histological response, recurrence and time to recurrence. A Cox regression analysis was conducted for the effects of neoadjuvant chemotherapy RDI, histological response, tumor location, gender and age on patient survival. Thirty medical records were analyzed. Survival, histological response, recurrence and time to recurrence were not affected by the chemotherapy RDI. The 5-year overall survival of the patient's population was found to be 63% with a median survival of 9.4 years. Patients with a good histological response had a longer survival than those with a bad response (mean survival times 11.0 vs. 6.6 years, log-rank test, P = 0.046). High DI is not a prognostic factor in osteosarcoma and maintaining it should not be a prime priority. Histological response is a prognostic but possibly not a reliable predictive factor, and further research is needed in order to find other reliable factors.

Osteosarcoma in pediatric patients and young adults: a single institution retrospective review of presentation, therapy, and outcome.

Background. Little is known about how cumulative chemotherapy delivery influences the poorer outcome observed in young adult (YA, 18–40 years) versus pediatric (<18 years) osteosarcoma patients. Here, we retrospectively examined differences in presentation, therapy, including cumulative chemotherapy dose, and outcome in YA and pediatric patients. Methods. We reviewed 111 cases of high-grade osteosarcoma at Moffitt Cancer Center between 1988 and 2012. Presentation factors, therapies, and survival were compared between YA and pediatric cohorts. Results. The cohorts were equivalent with respect to metastatic status, gender, tumor size, tumor site, and histological subtype. We found that the YA patients tended to have poorer histologic response to neoadjuvant chemotherapy measured by necrosis with 55% and 35% of pediatric versus YA patients responding favorably (P = 0.06). Only 39% of YA patients achieved the typical pediatric dose of methotrexate, doxorubicin, and cisplatin. These patients had a 3-year EFS of 76% (CI 53–100%) versus 47% (CI 26–69%; P = 0.09) in those who received less chemotherapy. Conclusion. Age continues to be a prognostic factor in osteosarcoma. Our study suggests that presentation factors are not associated with prognosis, while poorer response to chemotherapy and lower cumulative dose of chemotherapy delivered to YA patients may contribute to poorer outcomes.

XB130 Expression in Human Osteosarcoma: A Clinical and Experimental Study

Identifying prognostic factors for osteosarcoma (OS) aids in the selection of patients who require more aggressive management. XB130 is a newly characterized adaptor protein that was reported to be to be a prognostic factor of certain tumor types. However, the association between XB130 expression and the prognosis of OS remains unknown. In the present study, we investigated the association between XB130 expression and clinicopathologic features and prognosis in patients suffering osteosarcoma, and further investigated its potential role on OS cells in vitro and vivo. A retrospective immunohistochemical study of XB130 was performed on archival formalin-fixed paraffin-embedded specimens from 60 pairs of osteosarcoma and noncancerous bone tissues, and compared the expression of XB130 with clinicopathological parameters. We then investigate the effect of XB130 silencing on invasion in vitro and lung metastasis in vivo of the human osteosarcoma cell line. Immunohistochemical assays revealed that XB130 expression in osteosarcoma tissues was significantly higher than that in corresponding noncancerous bone tissues (P=0.001). In addition, high XB130 expression more frequently occurred in osteosarcoma tissues with advanced clinical stage (P=0.002) and positive distant metastasis (P=0.001). Moreover, osteosarcoma patients with high XB130 expression had significantly shorter overall survival and disease-free survival (both P<0.001) when compared with patients with the low expression of XB130. The univariate analysis and multivariate analysis shown that high XB130 expression and distant metastasis were the independent poor prognostic factor. We showed that XB130 depletion by RNA interference inhibited invasion of XB130-rich U2OS cells in vitro and lung metastasis in vivo. This is the first study to reveal that XB130 overexpression may be related to the prediction of metastasis potency and poor prognosis for osteosarcoma patients, suggesting that XB130 may serve as a prognostic marker for the optimization of clinical treatments. Furthermore, XB130 is the potential molecular target for osteosarcoma therapy.

Prognostic Value of Radiological Response to Chemotherapy in Patients with Osteosarcoma

BackgroundChemotherapy is essential to improve the prognosis of the patients with osteosarcoma, and the response to chemotherapy is an important prognostic factor. In this study, the impact of various radiological examinations on overall survival (OS) and event-free survival (EFS) was evaluated.MethodEighty-two patients with high-grade osteosarcoma were included in this study, and we evaluated the following factors for prognostic significance: age (≥40 years), gender (male), tumor location (truncal site), metastatic disease, histological response to chemotherapy, radiological response to chemotherapy assessed using X-ray, angiography, CT, MRI, 201Tl scintigraphy, and 99mTc-MIBI scintigraphy (99mTc-MIBI), and combined radiological score (CRS).ResultsUnivariate analyses revealed that metastatic disease, histological response, 99mTc-MIBI, and CRS were significantly correlated with OS. Multivariate analyses showed that metastatic disease (OS: HR 35.9, P<0.001; EFS: HR 17.32, P<0.001) was an independent predictor of OS and EFS. Tumor location (HR 36.1, P = 0.003), histological response (HR 31.1, P = 0.036), and 99mTc-MIBI (HR 18.4, P = 0.038) were significant prognostic factors for OS. Moreover, CRS was a marginally significant predictor of OS and EFS.ConclusionThe chemotherapeutic effects evaluated by 99mTc-MIBI and CRS could be considered as prognostic factors in osteosarcoma.

Proteomic approach toward molecular backgrounds of drug resistance of osteosarcoma cells in spheroid culture system

Chemoresistance is one of the most critical prognostic factors in osteosarcoma, and elucidation of the molecular backgrounds of chemoresistance may lead to better clinical outcomes. Spheroid cells resemble in vivo cells and are considered an in vitro model for the drug discovery. We found that spheroid cells displayed more chemoresistance than conventional monolayer cells across 11 osteosarcoma cell lines. To investigate the molecular mechanisms underlying the resistance to chemotherapy, we examined the proteomic differences between the monolayer and spheroid cells by 2D-DIGE. Of the 4762 protein species observed, we further investigated 435 species with annotated mass spectra in the public proteome database, Genome Medicine Database of Japan Proteomics. Among the 435 protein species, we found that 17 species exhibited expression level differences when the cells formed spheroids in more than five cell lines and four species out of these 17 were associated with spheroid-formation associated resistance to doxorubicin. We confirmed the upregulation of cathepsin D in spheroid cells by western blotting. Cathepsin D has been implicated in chemoresistance of various malignancies but has not previously been implemented in osteosarcoma. Our study suggested that the spheroid system may be a useful tool to reveal the molecular backgrounds of chemoresistance in osteosarcoma.

Role of integrin‐linked kinase in osteosarcoma progression

Although integrin-linked kinase (ILK) has been suggested to play a role in the tumorigenesis of a number of human epithelial carcinomas, little is known of its role in musculoskeletal sarcoma. The authors studied ILK expression by immunohistochemistry using osteosarcoma prechemotherapy specimens from 56 patients, and investigated the prognostic implications of the findings obtained. It was found that ILK overexpression was significantly correlated with the presence of distant metastasis (p = 0.008) and that it was an independent prognostic factor for both poor overall survival and poor event-free survival (p = 0.015 and 0.010, respectively). During a transfection experiment conducted by transfecting osteosarcoma cells with ILK siRNA, VEGF concentrations were measured using an ELISA kit, and then compared with those of untransfected controls to evaluate its angiogenic effects. In addition, apoptotic percentages were measured by Annexin-V flow cytometry, and invasive properties were evaluated by measuring the numbers of non-migrating cells in a Boyden chamber. It was found that ILK downregulation significantly decreased angiogenesis, increased apoptosis, and decreased invasiveness of osteosarcoma cells. These results show that ILK is a promising prognostic factor in osteosarcoma and a novel potential therapeutic target for the treatment of osteosarcoma.

Abstract 3815: Genomic predictors of response to preoperative chemotherapy in patients with pediatric osteosarcoma.

Abstract Purpose: It is generally accepted that sensitivity of preoperative chemotherapy, evaluated by a histological analysis using resected tumor tissue, is the most important prognostic factor for Osteosarcoma (OS). Methotrexate, doxorubicin, cisplatin (MAP regimen), and ifosfamide (IFO) are recognized as key drugs for OS, although it is still debatable whether addition of IFO for all OS patient is beneficial or not. There have been few analyses documenting molecular markers that predict the response to each regimen used in the preoperative chemotherapy of OS patients. The aim of this study is to identify possible genomic markers for the prediction of sensitivity of preoperative chemotherapy for pediatric OS patients. Methods: Pre-therapeutic biopsy tumor samples of 22 pediatric conventional OS patients treated in Chiba Cancer Center who had homogeneous clinicopathological- and therapeutic- background were analyzed. In our study, all patients were divided into 3 groups according to the histological response; group A includes 8 patients who demonstrated a good histological response to MAP regimen, group B includes 6 patients who were resistant to MAP regimen but showed good response to IFO regimen, and group C includes 8 patients who did not respond to neither MAP nor IFO regimen. Genomic DNAs were prepared from frozen biopsy samples and used for aCGH analysis with Agilent 44k DNA microarray. Results: Survival analysis revealed that patients in group A and B exhibited significantly better outcomes than in group C concerning both overall and event-free survivals. According to the results of comparative analysis of copy number variation (CNV) patterns in each group, 2 CNV regions that separated group A from group B and C (sensitivity 100%, specificity 92.9%) and 6 CNV regions that separated group C from group A and B (sensitivity 100%, specificity 100%) were identified. CNV regions specifically identified in Group C contained several genes previously reported as OS-related genes, such as CDKN2A/B, PTEN, and CDK4. Based on these CNV profiles, we established a scoring system that clearly classify the tumors into these 3 groups Re-sequencing of known cancer-related genes in these OS tumors is also underway to integrate genomic alterations into the system. Conclusion: Our results indicated that copy number changes in OS biopsy samples could be good predictors of response to preoperative chemotherapy. These signatures may be useful for the future genomics-based personalized chemotherapy for OS patients. Citation Format: Shintaro Iwata, Miki Ohira, Tsukasa Yonemoto, Takeshi Ishii, Hiroto Kamoda, Akira Nakagawara. Genomic predictors of response to preoperative chemotherapy in patients with pediatric osteosarcoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 3815. doi:10.1158/1538-7445.AM2013-3815

Suboptimal chemotherapy is an adverse prognostic factor in osteosarcoma

BackgroundWe sought to determine whether suboptimal chemotherapy compromised the prognosis of osteosarcoma patients.MethodsA total of 132 eligible patients who underwent chemotherapy between 1998 and 2008 were identified in our database. Information regarding patient demographics, clinical characteristics, and survival status were extracted for analysis. Optimal chemotherapy was defined as receipt of ≥80% of the planned dose intensity of prescribed agents within the planned durations.ResultsThe use of optimal chemotherapy resulted in an overall survival benefit with P = 0.006. Patients who failed to complete the optimal chemotherapy protocol had a dismal prognosis of 30.8% overall survival over five years, whereas those who completed the optimal chemotherapy had an overall survival rate over five years of 65.3%. Based on multivariate analysis, patients who were treated with a suboptimal protocol had a higher risk of relapse, metastasis and mortality. The hazard ratio (HR) of recurrence or death for the suboptimal chemotherapy group was as high as 2.512 over that of the optimal chemotherapy group (HR = 2.512, 95% confidence interval = 1.242 to 3.729).ConclusionsChemotherapy is a significant independent prognostic variable, and suboptimal chemotherapy was found to have a detrimental effect on the outcome of patients with osteosarcoma.

Age as a prognostic factor in osteosarcoma: Survival analysis.

10074 Background: Studies comparing survival between adult and paediatric population with osteosarcoma are scarce and contradictory. Generally adults were excluded from analysis in historical series. End Point: evaluate age as prognostic factor in ostosarcomas IIb treated exclusively by a multidisciplinary group in a single institution. Methods: 132/278 patients with histological diagnosis of osteosarcoma IIb were selected. All were treated exclusively in our hospital from July 1988 until December 2010. Patients 17 years old or younger were considered paediatric. They all received presurgical chemotherapy with the same scheme: ifosfamide + doxorrubicin + high dose methotrexate. Univariate analysis was made (Fischer exact test). Survival was calculated wih Kaplan-Meier actuarial method. Curves were compared with log rank test. Multivariate Cox analysis was made. Results: Median age was 19.6 years (std: 9,1; range 5-58). Adults: 77, children: 55. No differences were detected between the two age groups regarding: elevated alkaline phosphatase, kind of surgery (amputation vs. limb sparing), relapse site (lung, local, other), necrosis greater than 90% or number of lung resections. There was a tendency towards axial localization in adults (p=0.05). No paediatric patient had inadequate medical intervention, but it was present in 14.3% of adults (p=0.002). 5 year Overall survival (5y OS) in children was 85,2% compared with 61,8% in adults (p=0.005); Disease free survival (DFS) had a non significant tendency to be better in children (69.6% vs. 51,3%). Variables associated with worse OS were: axial location (p=0.01), elevated alkaline phosphatase (0.003), amputation (p=0.008), local relapse or systemic non-lung metastasis (p= 0.001), necrosis less than 90% (0.001). Multivariate Cox analysis showed association between OS and paediatric population (p=0.01) and necrosis greater than 90% (p=0.001), while DFS was associated with necrosis greater than 90% (p=0.01) and elevated alkaline phosphatase (p=0.05). Conclusions: Paediatric population presents better survival compared to adults in our institution. Differences in tumour biology and in the mode of presentation related to age may be the explanation.

Abstract 5579: Genomic approach to the prediction of chemotherapeutic sensitivity in osteosarcoma

Abstract PURPOSE: Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. Despite of recent progress of combined therapy, about 30% of OS patients die because of tumor progression or relapse. It is generally accepted that sensitivity of neoadjuvant chemotherapy is the most important prognostic factor for OS, however, there have been few analyses documenting molecular markers that predicts the chemosensitivity of OS patients. The aim of this study is to identify possible genomic biomarkers for the prediction of chemosensitivity and prognosis for OS patients. METHODS: Pretherapeutic biopsy samples of 22 pediatric OS patients treated in Chiba cancer center who had homogeneous clinicopathological- and therapeutic- background were analyzed. In our study, patients were divided into 3 groups; group A includes 8 patients who demonstrated a good response for 1st regimen, group B includes 6 patients who were resistant to 1st regimen but showed good response to 2nd regimen, group C includes 8 patients who did not respond to neither 1st nor 2nd regimen. Genomic DNAs were prepared from each frozen tissue and used for aCGH analysis with Agilent 44k DNA microarray. RESULTS: According to the results of comparative analysis of CNV patterns in each patient, three chromosomal patterns strongly separated group A from B&amp;C or C from A&amp;B were identified. Gain of 5p12 was found in 7 out of 8 patients of group A, while only 1 out of 14 patients of group B and C (sensitivity 87.5%, specificity 92.9%). On the other hand, loss of 9q21 and amplification of 12q14 were specifically observed in group C, but not in A and B (sensitivity 100%, specificity 92.9%). The average of copy numbers within the region of 5p12 and 9q21.3 on each patient were calculated, and 2-dimensional plot of these two values clearly showed that each group formed distinct clusters. CONCLUSION: Our results indicated that copy number changes in OS biopsy samples could be good markers to determine suitable chemotherapeutic regimen for individual patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5579. doi:1538-7445.AM2012-5579

High serum alkaline phosphatase cooperating with MMP-9 predicts metastasis and poor prognosis in patients with primary osteosarcoma in Southern China

BackgroundOsteosarcoma is a malignant tumor with high ability to form invasion and metastasis. Identifying prognostic factor in osteosarcoma is helpful to select those patients for more aggressive management. Our study evaluated serum alkaline phosphatase (ALP) cooperating with matrix metalloproteinase-9 (MMP-9) as an important prognostic predictor for local recurrence and distant metastasis of osteosarcoma.Methods177 cases were included from the osteosarcoma patients treated at 1st Affiliated Hospital of Sun Yat-sen University (1999-2008). Pre-chemotherapy serum ALP (pre-ALP) were studied and correlated with tumor recurrence, lung metastasis and patient survival. MMP-9 protein in tumor tissues was detected by immunohistochemistry and correlated with pre-ALP level.ResultsPre-ALP were partitioned into normal, high, and very high groups, in each group the incidence of metastases was 12.2%, 21.2% and 34.6%, respectively (p = 0.007). In the three groups the mean disease-free survival (DFS) was 57 ± 3.15, 28 ± 3.57 and 14 ± 3.35 months, respectively (p < 0.001); overall survival (OS) was 92 ± 26.89, 39 ± 8.61 and 17 ± 5.07 months, respectively (p < 0.001). By multivariate analysis, elevated serum pre-ALP were associated with shorter DFS (p = 0.018) and OS (p = 0.031). If elevated ALP levels decreased after clinical treatment, the incidence of lung metastasis rate decreased (p = 0.028); DFS and OS were both prolonged (p < 0.001). Pre-ALP was also positively correlated with MMP-9 expression (p = 0.015) in tumor tissue.ConclusionsPre-ALP was an independent prognostic factor for the survival of osteosarcoma patients in south China, and correlated with MMP-9 expression and lung metastasis. ALP can also serve as a prognostic marker for treatment, and merit large-scale validation studies.

Outcome for adolescent and young adult patients with osteosarcoma

There are conflicting data regarding age as a prognostic factor in osteosarcoma. The authors conducted a study evaluating the impact of age on prognosis in children and young adults with osteosarcoma enrolled on North American cooperative group trials. Patients with high-grade osteosarcoma of any site enrolled on North American cooperative group trials CCG-7943, POG-9754, INT-0133, and AOST0121 were included in this study. Primary tumor site, age, sex, ethnicity, histologic response, and presence of metastatic disease at diagnosis were evaluated for their impact on overall survival (OS) and event-free survival (EFS). A total of 1054 patients were eligible and had complete data available for the study. Age was not significantly associated with any other presenting covariate analyzed except sex. Age 18 or older was associated with a statistically significant poorer EFS (P = .019) and OS (P = .043). The 10-year EFS and OS in patients <10, 10 to 17, and ≥18 years old were 55%, 55%, 37% and 68%, 60%, 41%, respectively. The poorer EFS in patients ≥18 years old was because of an increased rate of relapse. Presence of metastatic disease at diagnosis, poor histologic response, and pelvic tumor site were also associated with a poorer prognosis. In multivariate analysis, age continued to be associated with poorer EFS (P = .019) and OS (P = .049). In osteosarcoma, age 18 to 30 years is associated with a statistically significant poorer outcome because of an increased rate of relapse. Poorer outcome in adolescent and young adult patients is not explained by tumor location, histologic response, or metastatic disease at presentation.

The ‘other’ bone sarcomas

We aimed to identify the incidence, outcome and prognostic factors associated with spindle cell sarcomas of bone (SCSB). We studied 196 patients with a primary non-metastatic tumour treated with the intent to cure. The results were compared with those of osteosarcoma patients treated at our hospital during the same period. The overall incidence of SCSB was 7.8% of all patients with a primary bone sarcoma. The five- and ten-year survival rates were 67.0% and 60.0%, respectively, which were better than those of patients with osteosarcoma treated over the same period. All histological subtypes had similar outcomes. On univariate analysis, factors that were significantly associated with decreased survival were age > 40 years, size > 8 cm, the presence of a pathological fracture, amputation, involved margins and a poor response to pre-operative chemotherapy. Multivariate analyses showed that age > 65 years, amputation and involved margins were all statistically significant prognostic factors. Involved margins and poor response to pre-operative chemotherapy were associated with an increased risk of local recurrence. SCSB has a better prognosis than osteosarcoma when matched for age. Most prognostic factors for osteosarcoma also seem to apply to SCSB. Patients with SCSB should be treated in the same way as patients of the same age with osteosarcoma.

Expression of matrix metalloproteinase regulator, RECK, and its clinical significance in osteosarcoma

Reversion-inducing cysteine-rich protein with Kazal motifs (RECK), a novel membrane-anchored matrix metalloproteinase inhibitor, have been shown to be associated with prognosis and suppress tumor progression through angiogenesis inhibition in many cancers. In this study, the expression of RECK in osteosarcoma was examined, and its clinical significance was firstly evaluated. RECK expression was immunohistochemically examined in osteosarcoma from 49 patients. By summing intensity and proportion scores, these patients were categorized as weak and strong. RECK expression in the primary tumor was strong in 27 patients (55.1%) and was weak in the rest of the patients. The 5-year survival rate of patients with RECK-strong tumor (81.5%) was significantly higher than that of patients with RECK-weak tumor (36.4%; p = 0.003). Reduced RECK expression significantly correlated with metastasis (p = 0.010) and recurrence (p = 0.004). A multivariate analysis confirmed that reduced RECK expression was an independent and significant factor to predict a poor prognosis (p = 0.017). RECK status is a useful prognostic factor in osteosarcoma, and an independent prognostic factor contributing to the determination of more adequate therapy strategies for each patient.

The clinical features and prognostic factors of osteosarcoma in adult.

Objective To study clinical features and outcomes of adult osteosareoma and evaluate prognostic factors for survival. Methods The clinical data of 30 over 40 years old of os-teosarcoma were retrospectively reviewed between March 2003 and March 2005, which included sex, age, tumor site, et al. Kaplan-Meier method was used to measure the overall survival rate. And log-rank univari-ate analysis was used to find prognostic factors related with survival rate. And Cox model multivariate analy-sis was used to find which prognostic factor was the independent one. Results All the were fol-lowed up at least 3 years, and 13 patient died with a median survival of 16months (range, 3-35 months) . Overall surviral rate was 56.7%. The tumors were at extremitities in 20 cases and axial bones in 10 cases. In X-ray film, 17 cases showed osteolytic lesions, 8 showed mixed, and 5 showed osteoblastic lesions. There were 22 cases with normal serum AKP levels, and 8 cases with higher levels of serum AKP. Overall surviral rate of 3 years was significantly related to patients age, tumor site, serum AKP level, Enneking surgical stage, surgical method and distant metastasis (Log-rank, P 0.05). Coulusion The clinical features of adult with osteosareoma was different from the young. There were more cases with tumor on axial bones in adult, and more cases with a normal serum AKP level, and osteolytic radiological finding. Surgical method played an important role in prognosis for survival in adult osteosarcoma patients. Key words: Adult; Osteosarcoma; Prognosis

VEGF expression as a prognostic marker in osteosarcoma

The vascular endothelial growth factor (VEGF) pathway is the key regulator of angiogenesis. In osteosarcoma baseline VEGF is of proven prognostic value but prognostic potential of post-NACT VEGF expression is largely unexplored. Treatment naive patients with osteosarcoma were subjected to initial staging workup followed by three cycles of neoadjuvant chemotherapy (NACT) and surgery; resected tumors were assessed for histological necrosis by Huvos grading. Initial biopsy and resected tumor specimens post-NACT were examined for VEGF expression by immunohistochemistry. Positive VEGF expression was considered when intensive positive staining was observed in >10% of the tumor cells. VEGF expression at baseline was compared with grade of tumor; pre-NACT and post-NACT VEGF expression were compared with histological necrosis. Receiver operating characteristic curves were generated to assess best threshold and predictability. A total of 31 patients were recruited with median age of 17 years (range 5-66 years); male/female ratio was 25:6; 23 patients (74%) were non-metastatic. At baseline, there was 90% concordance between positive VEGF expression and higher histological grade (28/31); baseline VEGF expression did not correlate well with stage and histological necrosis. Twenty-one (67%) were poor and 10 (33%) were good histologic responders; post-NACT VEGF expression as well as VEGF change following NACT significantly correlated with histological necrosis. Positive VEGF expression in surviving tumor cells post-NACT in resected tumors appears to be an important negative prognostic factor in osteosarcoma which may help future therapies to be identified according to the angiogenic potential of the disease.