Intra-abdominal tumors, such as ovarian cancer, have a clear predilection for metastasis to the omentum, an organ primarily composed of adipocytes. Currently, it is unclear why tumor cells ...preferentially home to and proliferate in the omentum, yet omental metastases typically represent the largest tumor in the abdominal cavities of women with ovarian cancer. We show here that primary human omental adipocytes promote homing, migration and invasion of ovarian cancer cells, and that adipokines including interleukin-8 (IL-8) mediate these activities. Adipocyte-ovarian cancer cell coculture led to the direct transfer of lipids from adipocytes to ovarian cancer cells and promoted in vitro and in vivo tumor growth. Furthermore, coculture induced lipolysis in adipocytes and β-oxidation in cancer cells, suggesting adipocytes act as an energy source for the cancer cells. A protein array identified upregulation of fatty acid-binding protein 4 (FABP4, also known as aP2) in omental metastases as compared to primary ovarian tumors, and FABP4 expression was detected in ovarian cancer cells at the adipocyte-tumor cell interface. FABP4 deficiency substantially impaired metastatic tumor growth in mice, indicating that FABP4 has a key role in ovarian cancer metastasis. These data indicate adipocytes provide fatty acids for rapid tumor growth, identifying lipid metabolism and transport as new targets for the treatment of cancers where adipocytes are a major component of the microenvironment.
Ovarian cancer (OvCa) metastasizes to organs in the abdominal cavity, such as the omentum, which are covered by a single layer of mesothelial cells. Mesothelial cells are generally thought to be ..."bystanders" to the metastatic process and simply displaced by OvCa cells to access the submesothelial extracellular matrix. Here, using organotypic 3D cultures, we found that primary human mesothelial cells secrete fibronectin in the presence of OvCa cells. Moreover, we evaluated the tumor stroma of 108 human omental metastases and determined that fibronectin was consistently overexpressed in these patients. Blocking fibronectin production in primary mesothelial cells in vitro or in murine models, either genetically (fibronectin 1 floxed mouse model) or via siRNA, decreased adhesion, invasion, proliferation, and metastasis of OvCa cells. Using a coculture model, we determined that OvCa cells secrete TGF-β1, which in turn activates a TGF-β receptor/RAC1/SMAD-dependent signaling pathway in the mesothelial cells that promotes a mesenchymal phenotype and transcriptional upregulation of fibronectin. Additionally, blocking α5 or β1 integrin function with antibodies reduced metastasis in an orthotopic preclinical model of OvCa metastasis. These findings indicate that cancer-associated mesothelial cells promote colonization during the initial steps of OvCa metastasis and suggest that mesothelial cells actively contribute to metastasis.
Tumor-associated macrophages (TAMs) exhibit an M2 macrophage phenotype that suppresses anti-tumor immune responses and often correlates with poor outcomes in patients with cancer. Patients with ...ovarian cancer frequently present with peritoneal carcinomatosis, but the mechanisms that induce naïve peritoneal macrophages into TAMs are poorly understood. In this study, we found an increased abundance of TAMs in mouse i.p. xenograft models of ovarian cancer that expressed HOXA9 , a homeobox gene that is associated with poor prognosis in patients with ovarian cancer. HOXA9 expression in ovarian cancer cells stimulated chemotaxis of peritoneal macrophages and induced macrophages to acquire TAM-like features. These features included induction of the M2 markers, CD163 and CD206, and the immunosuppressive cytokines, IL-10 and chemokine ligand 17, and down-regulation of the immunostimulatory cytokine, IL-12. HOXA9-mediated induction of TAMs was primarily due to the combinatorial effects of HOXA9-induced, tumor-derived transforming growth factor-β2 and chemokine ligand 2 levels. High HOXA9 expression in clinical specimens of ovarian cancer was strongly associated with increased abundance of TAMs and intratumoral T-regulatory cells and decreased abundance of CD8+ tumor-infiltrating lymphocytes. Levels of immunosuppressive cytokines were also elevated in ascites fluid of patients with tumors that highly expressed HOXA9. HOXA9 may, therefore, stimulate ovarian cancer progression by promoting an immunosuppressive microenvironment via paracrine effects on peritoneal macrophages.
Epithelial ovarian cancers (EOCs) often exhibit morphologic features of embryonic Müllerian duct-derived tissue lineages and colonize peritoneal surfaces that overlie connective and adipose tissues. ...However, the mechanisms that enable EOC cells to readily adapt to the peritoneal environment are poorly understood. In this study, we show that expression of HOXA9, a Müllerian-patterning gene, is strongly associated with poor outcomes in patients with EOC and in mouse xenograft models of EOC. Whereas HOXA9 promoted EOC growth in vivo, HOXA9 did not stimulate autonomous tumor cell growth in vitro. On the other hand, expression of HOXA9 in EOC cells induced normal peritoneal fibroblasts to express markers of cancer-associated fibroblasts (CAFs) and to stimulate growth of EOC and endothelial cells. Similarly, expression of HOXA9 in EOC cells induced normal adipose- and bone marrow-derived mesenchymal stem cells (MSCs) to acquire features of CAFs. These effects of HOXA9 were due in substantial part to its transcriptional activation of the gene encoding TGF-β2 that acted in a paracrine manner on peritoneal fibroblasts and MSCs to induce CXCL12, IL-6, and VEGF-A expression. These results indicate that HOXA9 expression in EOC cells promotes a microenvironment that is permissive for tumor growth.
Summary We report a detailed cytomorphologic evaluation of the circulating component of widely metastatic breast carcinoma. A previously healthy 38-year-old woman was diagnosed with breast cancer. ...Wide local excision revealed a 1.7-cm infiltrating ductal adenocarcinoma, BSR score 7/9 with angiolymphatic invasion, and 4/20 lymph nodes positive for carcinoma. Five years later, a bone marrow biopsy revealed involvement of bone marrow by metastatic breast carcinoma, and shortly thereafter, metastases were identified in the liver and lung hilum. She enrolled in a clinical investigation for the detection of circulating tumor cells (CTCs) in breast carcinoma. A total of 659 CTCs were identified in a 10-mL blood sample using an immunofluorescent protocol targeting cytokeratins and detected using fiber-optic array scanning technology. The detected CTCs were subsequently stained with a Wright-Giemsa stain, and representative cells were evaluated in detail by light microscopy for morphologic evaluation. We find that the patient's CTCs exhibit a high degree of pleomorphism including CTCs with high and low nuclear-to-cytoplasmic ratios along with CTCs exhibiting early and late apoptotic changes. In addition, in comparison with her tumor cells in other sites, the full morphologic spectrum of cancer cells present in primary and metastatic tumor is also present in peripheral blood circulation.
A Rare-Cell Detector for Cancer Krivacic, Robert T.; Ladanyi, Andras; Curry, Douglas N. ...
Proceedings of the National Academy of Sciences - PNAS,
07/2004, Letnik:
101, Številka:
29
Journal Article
Recenzirano
Odprti dostop
Although a reliable method for detection of cancer cells in blood would be an important tool for diagnosis and monitoring of solid tumors in early stages, current technologies cannot reliably detect ...the extremely low concentrations of these rare cells. The preferred method of detection, automated digital microscopy (ADM), is too slow to scan the large substrate areas. Here we report an approach that uses fiber-optic array scanning technology (FAST), which applies laser-printing techniques to the rare-cell detection problem. With FAST cytometry, laser-printing optics are used to excite 300,000 cells per sec, and emission is collected in an extremely wide field of view, enabling a 500-fold speed-up over ADM with comparable sensitivity and superior specificity. The combination of FAST enrichment and ADM imaging has the performance required for reliable detection of early-stage cancer in blood.
Purpose: Recently several reverse transcription-PCR techniques have been proven to be useful for the detection of circulating micrometastases.
However, this way intact cell clusters that were found ...in animal experiments of prognostic value could not be detected. In
this study, evaluation and modification of a commercial, cytokeratin-based, immunomagnetic cell separation method was performed
for the detection of intact cell clusters in colorectal carcinoma patients.
Experimental Design: Thirty-two colon cancer patients (6 were in Dukes stage B, 13 in stage C, and 13 in stage D) and 20 healthy donor samples
were evaluated. Immunomagnetic cell separation was performed from the buffy coat of peripheral blood samples (20 ml) using
the Carcinoma Cell Enrichment Kit (Miltenyi Biotec, Bergisch Gladbach, Germany), avoiding any filtering steps. The enriched
cell fraction was cytocentrifuged and immunocytochemically labeled using a pancytokeratin antibody (MNF116; Dako).
Results: Of 20 healthy samples, 2 contained one cytokeratin-positive cell. Of 32 single samples from malignant cases, 24 showed cytokeratin-positive
cells. Tumor cell clusters, mixed-cell doublets (one cytokeratin-positive and -negative cell), and mixed-cell clusters were
detected in 22 of 24 patients. In six cases, cytokeratin-positive dendritic-like cells were detected. Follow-up data indicate
that chemotherapy cannot destroy all of the circulating tumor cell clusters.
Conclusions: Using the methods presented, we could detect circulating colon cancer cells and cell clusters in colon carcinoma patients.
Similar cellular structures were described previously only in rats. Present data prove that such structures are present in
human colorectal cancer, too.
Ovarian cancer (OvCa) is characterized by widespread and rapid metastasis in the peritoneal cavity. Visceral adipocytes promote this process by providing fatty acids (FAs) for tumour growth. However, ...the exact mechanism of FA transfer from adipocytes to cancer cells remains unknown. This study shows that OvCa cells co-cultured with primary human omental adipocytes express high levels of the FA receptor, CD36, in the plasma membrane, thereby facilitating exogenous FA uptake. Depriving OvCa cells of adipocyte-derived FAs using CD36 inhibitors and short hairpin RNA knockdown prevented development of the adipocyte-induced malignant phenotype. Specifically, inhibition of CD36 attenuated adipocyte-induced cholesterol and lipid droplet accumulation and reduced intracellular reactive oxygen species (ROS) content. Metabolic analysis suggested that CD36 plays an essential role in the bioenergetic adaptation of OvCa cells in the adipocyte-rich microenvironment and governs their metabolic plasticity. Furthermore, the absence of CD36 affected cellular processes that play a causal role in peritoneal dissemination, including adhesion, invasion, migration and anchorage independent growth. Intraperitoneal injection of CD36-deficient cells or treatment with an anti-CD36 monoclonal antibody reduced tumour burden in mouse xenografts. Moreover, a matched cohort of primary and metastatic human ovarian tumours showed upregulation of CD36 in the metastatic tissues, a finding confirmed in three public gene expression data sets. These results suggest that omental adipocytes reprogram tumour metabolism through the upregulation of CD36 in OvCa cells. Targeting the stromal-tumour metabolic interface via CD36 inhibition may prove to be an effective treatment strategy against OvCa metastasis.
The aim of this study was to assess for an association between the sequence of surgical and systemic treatment and overall survival in patients with Stage IV endometrial cancer, as well as to ...identify prognostic factors for mortality.
The National Cancer Database (NCDB) was queried for cases of endometrial cancer diagnosed between 2006 and 2015. These cases were further narrowed to include only International Classification of Diseases for Oncology (ICD-O) histologic codes of adenocarcinoma or endometrioid adenocarcinoma and FIGO stage IV disease. Exclusion criteria were: reference date prior to date of diagnosis; unknown treatment sequence; intraoperative chemotherapy use; and hormonal or immunotherapy as first-line systemic therapy or unknown hormonal or immunotherapy use. Three groups were defined according to sequence of administration of surgery and systemic therapy. Overall survival (OS) was compared by calculating hazard ratios for death via Cox regression analysis. The Incomplete Treatment (IT) group received surgery alone, systemic therapy alone, or no treatment. The Primary Surgery (PS) group received surgery before systemic therapy. The Interval Surgery (IS) group had neoadjuvant systemic therapy followed by surgery, with or without further systemic therapy after surgery. Subanalyses were performed to assess the impact of post-IS systemic therapy, age, ethnicity, insurance status, and Charlson-Deyo score on OS. Analysis was performed using STATA.16.
A total of 8,520 cases of FIGO Stage IV endometrial adenocarcinoma were identified using the criteria above. Median OS in months for each treatment sequence group was 7.2 for IT, 35.7 for PS, and 30.7 for IS. Regardless of treatment sequence, receiving both surgical and systemic therapy improved OS compared to receiving only one or neither treatment. The hazard ratio (HR) for death for PS compared to IT was 0.38 (95% CI 0.36-0.40) and for IS compared to IT was 0.41 (95% CI 0.36-0.47), p<0.0001 for both. There was no significant difference in survival for IS compared to PS (HR 1.10, 95% CI 0.96-1.24, p=0.18). Stratification of IS cases by administration of further systemic therapy after surgery yielded the same result. Patient groups with worse OS (p<0.0001 for all) included: age >50 years (HR 1.18, 95% CI 1.08-1.30), non-Hispanic Black compared to non-Hispanic White (HR 1.27, 95% CI 1.17-1.37), and presence of medical comorbidities reflected by Charlson-Deyo score of 1 or more. Patients with private insurance had better OS than uninsured patients (HR 0.80, 95% CI 0.71-0.90, p<0.0001) but the same was not true for government insurance.
This retrospective survival analysis of NCDB data for FIGO Stage IV endometrial adenocarcinoma illustrated a benefit to receiving both systemic and surgical treatment over receiving only one or neither form of therapy. No difference in mortality was identified between primary surgery with adjuvant systemic therapy and neoadjuvant systemic therapy with interval surgery, even when accounting for additional chemotherapy after surgery. The decision whether to begin treatment with surgery or chemotherapy will thus depend on nuanced clinical factors specific to each patient. Patient factors associated with worse mortality included age >50 years, Non-Hispanic Black ethnicity, lack of insurance, and presence of medical comorbidities. These findings indicate a need to address the socioeconomic determinants of health contributing to these disparities in mortality.
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Purpose: The purpose of this study was to assess the feasibility of using rare event imaging system (REIS)-assisted analysis to detect
occult tumor cells (OTCs) in peripheral blood (PB). The study ...also sought to determine whether REIS-assisted OTC detection
presents a clinically viable alternative to manual microscopic detection to establish the true significance of OTC from solid
epithelial tumors.
Experimental Design: We recently demonstrated proof of concept using a fluorescence-based automated microscope system, REIS, for OTC detection
from the PB. For this study, the prototype of the system was adopted for high-throughput and high-content cellular analysis.
Results: The performance of the improved REIS was examined using normal blood ( n = 10), normal blood added to cancer cells ( n = 20), and blood samples obtained from cancer patients ( n = 80). Data from the screening of 80 clinical slides from breast and lung cancer patients, by manual microscopy and by the
REIS, revealed that as many as 14 of 35 positive slides (40%) were missed by manual screening but positively identified by
REIS. In addition, REIS-assisted scanning reliably and reproducibly quantified the total number of cells analyzed in the assay
and categorized positive cells based on their marker expression profile.
Conclusions: REIS-assisted analysis provides excellent sensitivity and reproducibility for OTC detection. This approach may enable an
improved method for screening of PB samples and for obtaining novel information about disease staging and about risk evaluation
in cancer patients.
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