Specific Metastatic Sites Research Articles

Novel immunohistochemistry-based signatures to predict metastatic site of triple-negative breast cancers

Background:Although distant metastasis (DM) in breast cancer (BC) is the most lethal form of recurrence and the most common underlying cause of cancer related deaths, the outcome following the development of DM is related to the site of metastasis. Triple negative BC (TNBC) is an aggressive form of BC characterised by early recurrences and high mortality. Athough multiple variables can be used to predict the risk of metastasis, few markers can predict the specific site of metastasis. This study aimed at identifying a biomarker signature to predict particular sites of DM in TNBC.Methods:A clinically annotated series of 322 TNBC were immunohistochemically stained with 133 biomarkers relevant to BC, to develop multibiomarker models for predicting metastasis to the bone, liver, lung and brain. Patients who experienced metastasis to each site were compared with those who did not, by gradually filtering the biomarker set via a two-tailed t-test and Cox univariate analyses. Biomarker combinations were finally ranked based on statistical significance, and evaluated in multivariable analyses.Results:Our final models were able to stratify TNBC patients into high risk groups that showed over 5, 6, 7 and 8 times higher risk of developing metastasis to the bone, liver, lung and brain, respectively, than low-risk subgroups. These models for predicting site-specific metastasis retained significance following adjustment for tumour size, patient age and chemotherapy status.Conclusions:Our novel IHC-based biomarkers signatures, when assessed in primary TNBC tumours, enable prediction of specific sites of metastasis, and potentially unravel biomarkers previously unknown in site tropism.

Abstract P6-09-21: Prognostic impact of metastatic pattern in stage IV breast cancer at initial diagnosis

Background: Stage IV breast cancer at initial diagnosis (BCID) can be recognized in approximately 5% of all breast neoplasms. Clinical outcomes of these patients (pts) are highly variable and depend on tumor biology and pt characteristics. The prognostic influence of metastatic pattern (MP) at initial presentation and factors associated with specific organ involvement have been understudied. The primary aim of this study was to analyze the influence of MP compared with other biologic and clinical factors in the survival of pts with stage IV BCID. The secondary aim was to evaluate factors associated with specific sites of metastatic spread. Methods: We evaluated women with microscopically confirmed stage IV BCID with known metastatic sites, reported to the Surveillance, Epidemiology and End Results (SEER) 18 registries program from 2010 to 2013. Pts with other primary tumor either before or after breast cancer were excluded. MP was categorized as bone only, visceral (lung, liver or brain), bone and visceral and other. Pt characteristics were compared between MP. Univariate and multivariate analyses determined the effects of each variable on overall survival (OS). Logistic regression examined factors associated with specific sites of metastases. Results: We included 9143 pts. Median age was 61 years (range 19-102). Median OS for the entire cohort was 28 months (95% CI 27-29 months). At diagnosis, bone only metastases represented 37.5% of pts, visceral 21.9%, bone and visceral 28.8% and other 11.9%. Median OS by MP was: bone only 38 months, visceral 21 months, bone and visceral 19 months and other 33 months (p Conclusions: To our knowledge, this is the largest study of MP in stage IV BCID. There were substantial differences in prognosis according to MP, bone only was the most common MP and had the best OS, whereas bone and visceral MP had the worst prognosis. We observed significant differences in pt characteristics according to MP. Independent predictors of OS included age at diagnosis, race, marital status, tumor grade, tumor subtype and MP. There was a clear influence of tumor subtype among other factors on specific sites of metastases. Our study identified several prognostic factors that could guide therapy selection in treatment naive pts.Background: Stage IV breast cancer at initial diagnosis (BCID) can be recognized in approximately 5% of all breast neoplasms. Clinical outcomes of these patients (pts) are highly variable and depend on tumor biology and pt characteristics. The prognostic influence of metastatic pattern (MP) at initial presentation and factors associated with specific organ involvement have been understudied. The primary aim of this study was to analyze the influence of MP compared with other biologic and clinical factors in the survival of pts with stage IV BCID. The secondary aim was to evaluate factors associated with specific sites of metastatic spread. Methods: We evaluated women with microscopically confirmed stage IV BCID with known metastatic sites, reported to the Surveillance, Epidemiology and End Results (SEER) 18 registries program from 2010 to 2013. Pts with other primary tumor either before or after breast cancer were excluded. MP was categorized as bone only, visceral (lung, liver or brain), bone and visceral and other. Pt characteristics were compared between MP. Univariate and multivariate analyses determined the effects of each variable on overall survival (OS). Logistic regression examined factors associated with specific sites of metastases. Results: We included 9143 pts. Median age was 61 years (range 19-102). Median OS for the entire cohort was 28 months (95% CI 27-29 months). At diagnosis, bone only metastases represented 37.5% of pts, visceral 21.9%, bone and visceral 28.8% and other 11.9%. Median OS by MP was: bone only 38 months, visceral 21 months, bone and visceral 19 months and other 33 months (p Conclusions: To our knowledge, this is the largest study of MP in stage IV BCID. There were substantial differences in prognosis according to MP, bone only was the most common MP and had the best OS, whereas bone and visceral MP had the worst prognosis. We observed significant differences in pt characteristics according to MP. Independent predictors of OS included age at diagnosis, race, marital status, tumor grade, tumor subtype and MP. There was a clear influence of tumor subtype among other factors on specific sites of metastases. Our study identified several prognostic factors that could guide therapy selection in treatment naive pts. Citation Format: Leone BA, Vallejo CT, Romero AO, Machiavelli MR, Perez JE, Leone J, Leone JP. Prognostic impact of metastatic pattern in stage IV breast cancer at initial diagnosis [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P6-09-21.

Prognostic impact of metastatic pattern in stage IV breast cancer at initial diagnosis.

To analyze the prognostic influence of metastatic pattern (MP) compared with other biologic and clinical factors in stage IV breast cancer at initial diagnosis (BCID) and evaluate factors associated with specific sites of metastases (SSM). We evaluated women with stage IV BCID with known metastatic sites, reported to the Surveillance, Epidemiology and End Results program from 2010 to 2013. MP was categorized as bone-only, visceral, bone and visceral (BV), and other. Univariate and multivariate analyses determined the effects of each variable on overall survival (OS). Logistic regression examined factors associated with SSM. We included 9143 patients. Bone represented 37.5% of patients, visceral 21.9%, BV 28.8%, and other 11.9%. Median OS by MP was as follows: bone 38months, visceral 21months, BV 19months, and other 33months (P<0.0001). Univariate analysis showed that higher number of metastatic sites had worse prognosis. In multivariate analysis, older age (hazard ratio 1.9), black race (hazard ratio 1.17), grade 3/4 tumors (hazard ratio 1.6), triple-negative (hazard ratio 2.24), BV MP (hazard ratio 2.07), and unmarried patients (hazard ratio 1.25) had significantly shorter OS. As compared with HR+/HER2- tumors, triple-negative and HR-/HER2+had higher odds of brain, liver, lung, and other metastases. HR+/HER2+had higher odds of liver metastases. All three subtypes had lower odds of bone metastases. There were substantial differences in OS according to MP. Tumor subtypes have a clear influence among other factors on SSM. We identified several prognostic factors that could guide therapy selection in treatment naïve patients.

Prognostic significance of pattern and burden of metastatic disease in patients with stage 4 neuroblastoma: A study from the International Neuroblastoma Risk Group database

Neuroblastoma is a childhood cancer with remarkably divergent tumour behaviour and the presence of metastatic disease is a powerful predictor of adverse outcome. However, the importance of the involvement of specific metastatic sites or overall metastatic burden in determining outcome has not been fully explored. We analysed data from the International Neuroblastoma Risk Group database for 2250 patients with stage 4 disease treated from 1990 to 2002. Metastatic burden was assessed using a ‘metastatic site index’ (MSI), a score based on the number of metastatic systems involved. Overall, involvement of bone marrow, bone, lung, central nervous system, or other sites was associated with worse outcome. For patients aged ≥18 months, involvement of liver had the greatest impact on outcome and was associated with tumour MYCN amplification and adrenal primary and lung metastases. Increased MSI was associated with worse outcome and higher baseline ferritin/lactate dehydrogenase. We explored the impact of initial treatment approach on these associations. Limiting the analysis to patients allocated to protocols including stem cell transplant (SCT), there was no longer an association of outcome with metastatic involvement of any individual system or increasing MSI. Thus, treatment escalation with SCT (and the addition of differentiating agents to maintenance therapy) appears to have provided maximal benefit to patients with greatest metastatic disease burden. These findings underscore the importance of examining prognostic factors in the context of specific treatments since the addition of new therapies may change or even negate the predictive impact of a particular variable.

Promoter hypermethylation profiling of distant breast cancer metastases.

Promoter hypermethylation of tumor suppressor genes seems to be an early event in breast carcinogenesis and is potentially reversible. This makes methylation a possible therapeutic target, a marker for treatment response and/or a prognostic factor. Methylation status of 40 tumor suppressor genes was compared between 53 primary breast tumors and their corresponding metastases to brain, lung, liver, or skin. In paired analyses, a significant decrease in methylation values was seen in distant metastases compared to their primaries in 21/40 individual tumor suppressor genes. Furthermore, primary tumors that metastasized to the liver clustered together, in line with the finding that primary breast carcinomas that metastasized to the brain, skin, or lung, showed higher methylation values in up to 27.5 % of tumor suppressor genes than primary carcinomas that metastasized to the liver. Conversion in methylation status of several genes from the primary tumor to the metastasis had prognostic value, and methylation status of some genes in the metastases predicted survival after onset of metastases. Methylation levels for most of the analyzed tumor suppressor genes were lower in distant metastases compared to their primaries, pointing to the dynamic aspect of methylation of these tumor suppressor genes during cancer progression. Also, specific distant metastatic sites seem to show differences in methylation patterns, implying that hypermethylation profiles of the primaries may steer site-specific metastatic spread. Lastly, methylation status of the metastases seems to have prognostic value. These promising findings warrant further validation in larger patient cohorts and more tumor suppressor genes.Electronic supplementary materialThe online version of this article (doi:10.1007/s10549-015-3362-y) contains supplementary material, which is available to authorized users.

Granulocytic immune infiltrates are essential for the efficient formation of breast cancer liver metastases.

IntroductionBreast cancer cells display preferences for specific metastatic sites including the bone, lung and liver. Metastasis is a complex process that relies, in part, on interactions between disseminated cancer cells and resident/infiltrating stromal cells that constitute the metastatic microenvironment. Distinct immune infiltrates can either impair the metastatic process or conversely, assist in the seeding, colonization and growth of disseminated cancer cells.MethodsUsing in vivo selection approaches, we previously isolated 4T1-derived breast cancer cells that preferentially metastasize to these organs and tissues. In this study, we examined whether the propensity of breast cancer cells to metastasize to the lung, liver or bone is associated with and dependent on distinct patterns of immune cell infiltration. Immunohistocytochemistry and immunohistofluorescence approaches were used to quantify innate immune cell infiltrates within distinct metastases and depletion of Gr1+ (Ly-6C and Ly-6G) or specifically Ly-6G+ cells was performed to functionally interrogate the role of Ly-6G+ infiltrates in promoting metastasis to these organs.ResultsWe show that T lymphocytes (CD3+), myeloid-derived (Gr-1+) cells and neutrophils (Ly-6G+ or NE+) exhibit the most pronounced recruitment in lung and liver metastases, with markedly less recruitment within bone metastatic lesions. Interestingly, these infiltrating cell populations display different patterns of localization within soft tissue metastases. T lymphocytes and granulocytic immune infiltrates are localized around the periphery of liver metastases whereas they were dispersed throughout the lung metastases. Furthermore, Gr-1+ cell-depletion studies demonstrate that infiltrating myeloid-derived cells are essential for the formation of breast cancer liver metastases but dispensable for metastasis to the lung and bone. A specific role for the granulocytic component of the innate immune infiltrate was revealed through Ly-6G+ cell-depletion experiments, which resulted in significantly impaired formation of liver metastases. Finally, we demonstrate that the CD11b+/Ly-6G+ neutrophils that infiltrate and surround the liver metastases are polarized toward an N2 phenotype, which have previously been shown to enhance tumor growth and metastasis.ConclusionsOur results demonstrate that the liver-metastatic potential of breast cancer cells is heavily reliant on interactions with infiltrating Ly-6G+ cells within the liver microenvironment.Electronic supplementary materialThe online version of this article (doi:10.1186/s13058-015-0558-3) contains supplementary material, which is available to authorized users.

Divergent and convergent evolution in metastases suggest treatment strategies based on specific metastatic sites.

Systemic therapy for metastatic cancer is currently determined exclusively by the site of tumor origin. Yet, there is increasing evidence that the molecular characteristics of metastases significantly differ from the primary tumor. We define the evolutionary dynamics of metastases that govern this molecular divergence and examine their potential contribution to variations in response to targeted therapies. Darwinian interactions of transformed cells with the tissue microenvironments at primary and metastatic sites are analyzed using evolutionary game theory. Computational models simulate responses to targeted therapies in different organs within the same patient. Tumor cells, although maximally fit at their primary site, typically have lower fitness on the adaptive landscapes offered by the metastatic sites due to organ-specific variations in mesenchymal properties and signaling pathways. Clinically evident metastases usually exhibit time-dependent divergence from the phenotypic mean of the primary population as the tumor cells evolve and adapt to their new circumstances. In contrast, tumors from different primary sites evolving on identical metastatic adaptive landscapes exhibit phenotypic convergence. Thus, metastases in the liver from different primary tumors and even in different hosts will evolve toward similar adaptive phenotypes. The combination of evolutionary divergence from the primary cancer phenotype and convergence towards similar adaptive strategies in the same tissue cause significant variations in treatment responses particularly for highly targeted therapies. The results suggest that optimal therapies for disseminated cancer must take into account the site(s) of metastatic growth as well as the primary organ.

Liver metastasis predicts poorer prognosis in stage IV lung adenocarcinoma patients receiving first-line gefitinib.

Gefitinib is currently used as a first-line therapy in patients of advanced non-small cell lung cancer (NSCLC) with susceptible epidermal growth factor receptor (EGFR) mutations. However, treatment outcomes of these patients vary. This study was conducted to evaluate the impact of specific metastatic sites on treatment outcomes of patients with stage IV lung adenocarcinoma with susceptible EGFR mutations receiving first-line gefitinib, focusing on the impact of liver metastasis. Between October 2009 and April 2014, patients of stage IV lung adenocarcinoma harboring EGFR mutation in exon 19 or 21, who received first-line gefitinib treatment, were enrolled in two hospitals and followed until December 22, 2014. The impacts of various clinical features, including sex, age, smoking history, performance status, EGFR mutation site, metastatic sites, etc., on progression-free survival (PFS) and overall survival (OS) were analyzed. A total of 148 patients were eligible for analysis. Patients with liver metastasis on initial diagnosis (n=19) had shorter PFS and OS than those without liver metastasis did (median of PFS, 6.7 vs. 11.2 months, p<0.0001; median of OS, 9.2 vs. 17.5 months, p<0.0001). Multivariable Cox regression analysis showed liver metastasis was an independent poor prognostic factor for PFS (HR=2.939 [95% CI: 1.729-4.997], p<0.0001) and OS (HR=3.300 [95% CI: 1.708-6.373], p=0.0004). Liver metastasis predicts poorer PFS and OS in stage IV lung adenocarcinoma patients with susceptible gene mutations receiving first-line gefitinib. Further study is warranted to elucidate the underlying mechanisms and find treatment modalities to improve prognosis of these patients.

Complete loss of PTEN protein expression correlates with shorter time to brain metastasis and survival in stage IIIB/C melanoma patients with BRAFV600 mutations.

Loss of function of PTEN is a frequent event in melanoma, particularly in tumors with BRAF(V600) mutations. The prevalence, pathologic features, and clinical outcomes associated with PTEN loss in patients with stage IIIB/C melanoma were interrogated to improve our understanding of the clinical significance of this molecular event. Archival tissue from lymphadenectomy specimens among patients (n = 136) with stage IIIB or IIIC melanoma was assessed by DNA sequencing for activating BRAF and NRAS mutations, and by immunohistochemistry for the expression of PTEN protein. Associations of these molecular aberrations with demographics, tumor characteristics, and clinical outcomes were determined. The prevalence of BRAF(V600) mutations (40% overall), NRAS mutations (10%), and PTEN loss (25%) did not vary by pathologic substage. BRAF/NRAS mutation status did not correlate with distant disease-free survival (DDFS) or overall survival (OS). Complete loss of PTEN expression correlated with shorter OS but not DDFS. When stratified by specific sites of distant metastasis, PTEN loss was associated with significantly shorter time to melanoma brain metastasis (MBM), but not to liver, lung, or bone metastasis. Analysis of PTEN in mutationally defined subsets showed that PTEN loss was significantly associated with OS and time to MBM in patients with BRAF(V600) mutations. Loss of PTEN protein expression correlates significantly with decreased OS and time to MBM in stage IIIB/C melanoma patients with BRAF(V600) mutations. The findings add to evidence supporting a significant role for PTEN loss and the PI3K-AKT pathway in melanoma.

Abstract 52: Identification of genomic alterations associated with metastasis in clear cell renal cell carcinoma

Abstract Clear cell renal cell carcinoma (ccRCC) is the most abundant and lethal form of RCC and about one third of patients exhibit metastasis with poor prognosis. At the molecular level, metastasis is accompanied by multiple genetic and epigenetic alterations including genomic copy number changes. Identification of novel genomic markers and underlying mechanisms associated with metastasis could result in better therapeutic management of ccRCC patients. In this study, aCGH was performed using Agilent 244K arrays on DNA from 83 primary and 64 metastatic unmatched ccRCC fresh-frozen specimens and profiles were compared using Fisher's Exact test (FDR, p-value ≤ 0.05), with a minimum threshold difference of 15%. Significant copy number changes were found at 15 loci between primary and metastatic lesions, after excluding loci enriched in normal CNVs. Loss of 2p16.1-p15, 2q33.1-q33.3, 3q11.1-q13.2, 3q22.3, 10q21.3-q22.1 and 11p15.4 were more frequent in primary lesions, while nine aberrations (gain of 5q23.3-q31.1, 7q21.3-q22.1, 16p11.2, 20q11.1-q13.12, 21p11.2-q11.1 and loss of 4p15.2-p14, 4q, 9p24.3-p13.3 and 18q21.2-q21.31) were more frequent in metastatic lesions. While some of these aberrations have previously been implicated in metastasis, nine were novel. In order to further examine the association of these aberrations with metastasis, the frequencies of the 15 aberrations were determined in primary ccRCCs publicly available from TCGA, stratified into low (stages I and II, n=287) and high stage groups (III and IV, n=202). Eight of the nine aberrations that were more frequent in metastatic lesions, were also observed at a higher frequency in high stage TCGA primary ccRCC (four were significantly higher between Stages I and IV). Little or no trend was observed for the six aberrations more frequent in primary lesions. In order to identify alterations associated with specific metastatic sites, aCGH profiles were compared across metastatic lesions at sites represented by at least 6 specimens: 15 lung, 7 bone, 6 lymph node, 6 pancreas and 6 brain metastatic ccRCC specimens. Gain of 7q11 and 8p12-p11, and loss of 6q14-q22 and 10q were specifically observed in lung metastases. Gain of chr12 and loss of chr22 were frequent in bone, loss of 1q32-q41 and gain of 22q11 in lymph node, and loss of 2p22.3, 8p12, 8q12-q13 and 8q24 in pancreatic lesions. No specific aberrations were found in brain lesions. In conclusion, this study has confirmed known and identified novel genomic loci associated with metastasis in ccRCC. Determination of candidate genes of the aberrations will further our understanding of metastatic progression of ccRCC and together with examination of metastatic site-specific alterations, may assist in selecting patients with metastatic disease for appropriate targeted therapy and lead to improved overall survival. Citation Format: Venkata J. Thodima, Banumathy Gowrishankar, Ana Molina, Murielle Georges, RSK Chaganti, Robert J. Motzer, Jane Houldsworth. Identification of genomic alterations associated with metastasis in clear cell renal cell carcinoma. [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 52. doi:10.1158/1538-7445.AM2014-52

Ras in digestive oncology: from molecular biology to clinical implications.

The modalities of Ras mutation detection, its role as a predictive biomarker, mechanisms of wild-type Ras activation, and the role of Ras-directed targeted therapies will be discussed mainly in colorectal cancer. RAS genotype is generally considered to be highly concordant between primary colorectal tumours and metastases. However, recent data show significant discordance between primary tumours and specific metastatic sites, but also heterogeneity within primary tumours. Moreover, the mechanisms of Ras activation expand far beyond mutations through altered expression or function of physiological Ras activators and inhibitors. Accordingly, genomic signatures of Ras or epidermal growth factor receptor (EGFR) activation are being developed and are potential predictive biomarkers of response to anti-EGFR antibodies. Finally, several recent clinical trials targeting Ras or its downstream signalling with statins or Raf inhibitors have shown promising activity in chemorefractory metastatic colorectal cancer. RAS mutation remains an important biomarker predicting response to anti-EGFR therapies and perhaps clinical outcomes after surgery for metastatic colorectal cancer, but new techniques including genomic signatures need to be validated to take into account the complexity of Ras activation. The importance of Ras signalling as a therapeutic target has recently been outlined by successful clinical trials with Raf inhibitors.

Metastasis mechanism and gene/protein expression in gastric cancer with distant organs metastasis.

Metastasis is the most fatal characteristics of malignancy tumor, which accounted for more than 90% of tumor-related mortality. Distant organ or tissue metastasis is a sign of poor prognosis in patients with gastric cancer. Tumor cells metastasis is a very complex process including tumor cell transformation, growth, angiogenesis, invasion, dissemination and survival in the circulation, and subsequent adhesion and colonization the secondary organ or tissue. The origin of tumor cell, genetic variation, the circulatory mode and the physiological structure of the metastatic organ determines the specific sites of distant metastasis. In theory, the metastatic lesion is originated from their primary tumor, so they should have the same molecular profile with the primary tumor. But this view has been confirmed to be wrong in various tumors, including gastric cancer. The gene expression of primary gastric cancer and its metastasis have differences, which may contribute to the early diagnosis and individualized treatment of metastasis. However, the heterogeneity of tumor cells is still unclear in different metastasis lesion of gastric cancer, which will be a major focus of future research. In this review, we discuss the basic principles of cancer metastasis, the unique physiological characteristics of the various metastasis organs and the expression of different functions of gene/protein in primary and metastasis of gastric cancer. In addition, we also discuss the diagnosis, prognosis and treatment in various organ metastasis of gastric cancer.

Abstract C35: The development of metastatic mouse models of neuroblastoma to bone and brain to identify the molecular mechanisms governing metastasis

Abstract Metastasis is a key reason for the lack of success in treating children with neuroblastoma (NB). Over 50% of patients present with metastatic lesions in the bone, brain, liver and lymph nodes, with a high fatality rate for patients that relapse with those metastases. A better understanding of the molecular processes defining the complexity and multi-step nature of NB dissemination is required for the development of effective treatments. To date, there is no NB model that quantitatively and reproducibly metastasizes to bone and brain. Thus, we developed novel metastatic mouse models of neuroblastoma to bone and brain through in vivo selection to enhance metastatic capability. SK-N-AS, derived from a bone marrow metastasis, and IMR-32, a MYCN-amplified line derived from an abdominal mass, were tagged for in vivo monitoring using a triple reporter (TR) encoding TK, GFP and firefly luciferase, which allows for nuclear imaging, FACs analysis, and whole body bioluminescence. Reporter expression was confirmed by double sorting for GFP and testing for luciferase activity. Cells were introduced into the blood stream of NOD/SCID mice through intra-cardiac injection, and monitored bi-weekly using bioluminescence. SK-N-ASTR cells metastasized to the adrenal gland (87%, 13/15 animals) and bone (mandible, hindlimbs, 67%, 10/15 animals), whereas IMR-32TR cells metastasized to bone (100%, 10/10 animals) and brain (40%, 4/10 animals). Metastatic lesions were confirmed by ex-vivo bioluminescence, 3D bioluminescence, MRI or micro-CT. Cells from specific metastatic sites of individual mice were isolated, expanded in culture, GFP sorted and injected into a second cohort of mice. Several of the SK-N-ASTR re-injected cell lines derived from bone exhibited enhanced metastasis after one round of in vivo selection, with 100% metastasis to bone and the adrenal gland (a common location of primary NB tumours) and up to 60% metastasis to brain, as confirmed by MRI. Subsequently, these animals had a decreased overall survival rate of approximately 43 days relative to the parental cell line, of 69 days. The metastatic sub-populations do not proliferate at higher rates than the parental cell line, as assessed by BrdU labeling. Thus, the enhanced metastasis observed in vivo is not due to a faster growth rate, but rather selection of metastasis promoting functions. This is supported by the finding that the metastatic SK-N-ASTR sub-populations also exhibited enhanced migration and invasion toward serum, as assessed using boyden chambers, compared to the parental line. These sub-populations are being characterized to identify the molecular mechanisms governing metastasis. Metastasis to brain and bone is an often fatal event in children with newly diagnosed and recurrent NB. We have developed the first reproducible and efficient NB metastatic mouse model to identify novel genes and signalling proteins regulating metastasis, and to identify drug candidates that suppress metastatic growth. Citation Format: Kelly E. Fathers, Samar Mouaaz, Constanza Rioseco, Anna Mourskaia, Ronald Blasberg, Meredith Irwin, Peter Siegel, David Kaplan. The development of metastatic mouse models of neuroblastoma to bone and brain to identify the molecular mechanisms governing metastasis. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C35.

The Tumor-Promoting Flow of Cells Into, Within and Out of the Tumor Site: Regulation by the Inflammatory Axis of TNFα and Chemokines

Tumors are dynamic organs, in which active processes of cell motility affect disease course by regulating the composition of cells at the tumor site. While sub-populations of tumor-promoting leukocytes are recruited inward and endothelial cell migration stands in the basis of vascular branching throughout the tumor, cancer cells make their way out of the primary site towards specific metastatic sites. This review describes the independent and cross-regulatory roles of inflammatory chemokines and of the inflammatory cytokine tumor necrosis factor α (TNFα) in determining cell motility processes that eventually have profound effects on tumor growth and metastasis. First, the effects of inflammatory chemokines such as CCL2 (MCP-1), CCL5 (RANTES) and CXCL8 (IL-8) are described, regulating the inward flow of leukocyte sub-populations with pro-tumoral activities, such as tumor-associated macrophages (TAM), myeloid-derived suppressor cells (MDSC), tumor-associated neutrophils (TAN), Th17 cells and Tregs. Then, the ability of inflammatory chemokines to induce endothelial cell migration, sprouting and tube formation is discussed, with its implications on tumor angiogenesis. This part is followed by an in depth description of the manners by which TNFα potentiates the above activities of the inflammatory chemokines, alongside with its ability to directly induce migratory processes in the tumor cells thus promoting metastasis. Note worthy is the ability of TNFα to induce in the tumor cells the important process of epithelial-to-mesenchymal transition (EMT). Emphasis is given to the ability of TNFα to establish an inflammatory network with the chemokines, and in parallel to form a cell re-modeling network together with transforming growth factor β (TGFβ). The review concludes by discussing the implications of such networks on disease course, and on the future design of therapeutic measures in cancer.

Glutamate receptors: emerging players in melanomagenesis

Glutamate receptors: emerging players in melanomagenesis

To build a prognostic score model containing indispensible tumour markers for metastatic nasopharyngeal carcinoma in an epidemic area

Background and objectiveThe survival outcomes of patients with metastatic nasopharyngeal carcinoma (NPC) differ significantly between individuals. The aim of this study is to build a prognostic score model (PSM) incorporating circulating tumour markers for metastatic NPC in an epidemic area. MethodsSeven hundred and ninety-nine patients with disseminated NPC were analysed retrospectively. Univariate and multivariable analyses were conducted using the Cox proportion hazards model. Factors analysed included patients, characteristics (gender, age group, performance status), circulating tumour-marker characteristics (Epstein–Barr virus (EBV) DNA level, EBV VCA-IgA level, lactate dehydrogenase (LDH) level, alkaline phosphatase (ALP) level), basic laboratory characteristics (leucocyte count, haemoglobin level, albumin level), and disease characteristics (presence of metastasis at presentation, disease-free interval, number of metastatic sites, specific metastatic sites). The PSM was built according to numerical score derived from the regression coefficients of each independent prognostic variable. The prognostic score of each patient was calculated by totalling up the scores of each independent variable. ResultsIndependent prognostic factors included performance status, age, haemoglobin level, LDH level, ALP level and EBV DNA level. Three prognostic groups based on PSM were obtained: low risk (total score=0–4); intermediate risk (5–8); high risk (9–12). Median survivals of the three groups were 25.5, 15.1 and 7months, respectively, (P<0.001). ConclusionClinical and laboratory characteristics can help guide the prognostication of patients with metastatic NPC in epidemic areas.

Genomic signatures of specific sites of metastasis

The conventional approach to cancer therapy, to treat according to the organ or tissue of origin and patients’ demographics, is gradually being replaced by a more personalized approach in which treatment choice is based on detailed knowledge of the genetic defects that underlie the oncogenic process in each individual tumor. As a result, we have witnessed the beginning of a shift from broadly-acting cytotoxic drugs towards more specific and less toxic targeted therapies. At the same time, the annotation of the human genome has enabled the development of a new class of gene expression-based molecular diagnostics that help to guide the choice of therapy.

Prognostic factors (PF) influencing overall survival (OS) in stage IV colorectal cancer (CRC).

e14021 Background: Ample data exists on prognostic factors in relapsed CRC, highlighting the importance of stage, time to recurrence, and adjuvant therapy. PFs among patients with de novo stage IV CRC are less well defined and may differ from those seen in relapsed disease. Methods: To evaluate the prognostic value of clinical and pathological factors, a retrospective review of 2,050 patients diagnosed from 1999 to 2007 and referred to the British Columbia Cancer Agency (BCCA) was undertaken. Patients were included in the study if newly referred to the BCCA, and the presence of stage IV disease was confirmed within four months of initial CRC diagnosis. OS was the primary endpoint and PFs were analyzed using Cox proportional hazards models. Results: 2,050 patients were included with a median age of 66. Palliative chemotherapy was given in 69% of patients. Many patients (1,435/2,050, 70%) had the primary tumor resected, which was associated with improved OS when compared against unresected primaries (HR 0.46 [0.41, 0.51], p<0.0001). Multivariate analysis was conducted on patients with resected primary tumors (Table). Tumor site (colon versus rectal), tumor (T) stage and lymphovascular invasion were not prognostic. Conclusions: Nodal (N) stage, particularly N2 disease, and tumor grade are prognostic among patients with primary resected stage IV CRC. Other prognostic factors include age, ECOG status, and specific sites of metastasis. Variable P value HR [95% CI] N1 vs N0 0.036 1.22 [1.01, 1.47] N2 vs N0 <0.0001 1.51 [1.26, 1.80] Tumor grade 3-4 vs 1-2 <0.0001 1.54 [1.35, 1.76] Site of mets (lung only vs liver only) 0.03 0.74 [0.57, 0.97] Site of mets (other solitary vs liver only) 0.052 0.83 [0.68, 1.00] Site of mets (multiple site vs liver only) <0.0001 1.45 [1.26, 1.68] Age >70 vs ≤70 <0.0001 1.36 [1.20, 1.54] ECOG 2 vs 0-1 <0.0001 1.56 [1.31, 1.85] ECOG 3-4 vs 0-1 <0.0001 2.98 [2.44, 3.63]

Retrospective study of the effect of disease progression on patient reported outcomes in HER-2 negative metastatic breast cancer patients

BackgroundThis retrospective study evaluated the impact of disease progression and of specific sites of metastasis on patient reported outcomes (PROs) that assess symptom burden and health related quality of life (HRQoL) in women with metastatic breast cancer (mBC).MethodsHER-2 negative mBC patients (n = 102) were enrolled from 7 U.S. community oncology practices. Demographic, disease and treatment characteristics were abstracted from electronic medical records and linked to archived Patient Care Monitor (PCM) assessments. The PCM is a self-report measure of symptom burden and HRQoL administered as part of routine care in participating practices. Linear mixed models were used to examine change in PCM scores over time.ResultsMean age was 57 years, with 72% of patients Caucasian, and 25% African American. Median time from mBC diagnosis to first disease progression was 8.8 months. Metastasis to bone (60%), lung (28%) and liver (26%) predominated at initial metastatic diagnosis. Results showed that PCM items assessing fatigue, physical pain and trouble sleeping were sensitive to either general effects of disease progression or to effects associated with specific sites of metastasis. Progression of disease was also associated with modest but significant worsening of General Physical Symptoms, Treatment Side Effects, Acute Distress and Impaired Performance index scores. In addition, there were marked detrimental effects of liver metastasis on Treatment Side Effects, and of brain metastasis on Acute Distress.ConclusionsDisease progression has a detrimental impact on cancer-related symptoms. Delaying disease progression may have a positive impact on patients' HRQoL.

CXCR4 Regulates the Early Extravasation of Metastatic Tumor Cells In Vivo

Recent studies have demonstrated that the chemokine receptor CXCR4 plays a crucial role in organ-specific metastasis formation. Although a variety of studies showed the expression of chemokine receptors, in particular, CXCR4, by gastrointestinal tumors, the precise mechanisms of chemokine receptor-mediated homing of cancer cells to specific sites of metastasis remained elusive. Here, we used liver metastatic human HEP-G2 hepatoma and HT-29LMM colon cancer cells expressing functional CXCR4 to dissect the metastatic cascade by intravital fluorescence microscopy. Immunohistochemistry revealed that the CXCR4 ligand CXCL12 is expressed by endothelial cells and likely Kupffer cells lining the liver sinusoids. Tumor cell adhesion and extravasation in vivo was quantitatively analyzed using intravital fluorescence microscopy. Treatment of cells with an anti-CXCR4 antibody did not affect cell adhesion but significantly impaired tumor cell extravasation (HEP-G2; isotype control: 22.3% ± 4.3% vs anti-CXCR4: 6.0% ± 5.0%, P < .001). In addition, pretreatment of tumor cells with the ligand CXCL12 enhanced the activation of the small GTPases Rho, Rac, and cdc42 as well as tumor cell extravasation without affecting tumor cell adhesion within liver sinusoids. Taken together, the findings of the present study provide first in vivo insights into the early events of chemokine ligand/receptor-mediated liver metastasis formation of tumor cells and define tumor cell extravasation rather than tumor cell arrest as the rate-limiting event.